                         SEQUENCE LISTING

<110>  YEDA RESEARCH AND DEVELOPMENT CO. LTD.
       FUTERMAN, Anthony H.
       SILMAN, Israel
       SUSSMAN, Joel L.
       FLEISHMAN, Sarel
       GOLDENZWEIG, Adi
       POKORNA, Sarka
       ASHANI, Yaacov
       KHERSONSKY, Olga
 
<120>  VARIANTS OF BETA-GLUCOCEREBROSIDASE FOR USE IN TREATING GAUCHER 
       DISEASE

<130>  86504

<150>  IL 273684
<151>  2020-03-29

<150>  US 63/049,685
<151>  2020-07-09

<160>  29    

<170>  PatentIn version 3.5

<210>  1
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  NA sequence of WT Gcase (E.coli optimized codons)

<400>  1
gcgcgcccgt gcattccgaa aagctttggc tatagcagcg tggtgtgcgt gtgcaacgcg       60

acctattgcg atagctttga tccgccgacc tttccggcgc tgggcacctt tagccgctat      120

gaaagcaccc gcagcggccg ccgcatggaa ctgagcatgg gcccgattca ggcgaaccat      180

accggcaccg gcctgctgct gaccctgcag ccggaacaga aatttcagaa agtgaaaggc      240

tttggcggcg cgatgaccga tgcggcggcg ctgaacattc tggcgctgag cccgccggcg      300

cagaacctgc tgctgaaaag ctattttagc gaagaaggca ttggctataa cattattcgc      360

gtgccgatgg cgagctgcga ttttagcatt cgcacctata cctatgcgga taccccggat      420

gattttcagc tgcataactt tagcctgccg gaagaagata ccaaactgaa aattccgctg      480

attcatcgcg cgctgcagct ggcgcagcgc ccggtgagcc tgctggcgag cccgtggacc      540

agcccgacct ggctgaaaac caacggcgcg gtgaacggca aaggcagcct gaaaggccag      600

ccgggcgata tttatcatca gacctgggcg cgctattttg tgaaatttct ggatgcgtat      660

gcggaacata aactgcagtt ttgggcggtg accgcggaaa acgaaccgag cgcgggcctg      720

ctgagcggct atccgtttca gtgcctgggc tttaccccgg aacatcagcg cgattttatt      780

gcgcgcgatc tgggcccgac cctggcgaac agcacccatc ataacgtgcg cctgctgatg      840

ctggatgatc agcgcctgct gctgccgcat tgggcgaaag tggtgctgac cgatccggaa      900

gcggcgaaat atgtgcatgg cattgcggtg cattggtatc tggattttct ggcgccggcg      960

aaagcgaccc tgggcgaaac ccatcgcctg tttccgaaca ccatgctgtt tgcgagcgaa     1020

gcgtgcgtgg gcagcaaatt ttgggaacag agcgtgcgcc tgggcagctg ggatcgcggc     1080

atgcagtata gccatagcat tattaccaac ctgctgtatc atgtggtggg ctggaccgat     1140

tggaacctgg cgctgaaccc ggaaggcggc ccgaactggg tgcgcaactt tgtggatagc     1200

ccgattattg tggatattac caaagatacc ttttataaac agccgatgtt ttatcatctg     1260

ggccatttta gcaaatttat tccggaaggc agccagcgcg tgggcctggt ggcgagccag     1320

aaaaacgatc tggatgcggt ggcgctgatg catccggatg gcagcgcggt ggtggtggtg     1380

ctgaaccgca gcagcaaaga tgtgccgctg accattaaag atccggcggt gggctttctg     1440

gaaaccatta gcccgggcta tagcattcat acctatctgt ggcatcgcca gtaa           1494


<210>  2
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AA sequence of WT Gcase

<400>  2

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Leu Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn His Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asn Leu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Ile Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


His Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Leu Gln Leu Ala Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Ile Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Arg Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Glu His Lys 
    210                 215                 220                 


Leu Gln Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Leu Ser Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asp Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Thr Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Thr Asn Leu Leu Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ile Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Leu Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Lys Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp His Arg 
                485                 490                 495     


Gln 
    


<210>  3
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified -glucocerebrosidase  (GCase variant)

<400>  3
gcgcgcccgt gcattccgaa aagctttggc tatagcagcg tggtgtgcgt gtgcaacgcg       60

acctattgcg atagctttga tccgccgacc tttccggcgc tgggcacctt tagccgctat      120

gaaagcaccc gcagcggccg ccgcatggaa ctgagcatgg gcccgattca ggcgaaccat      180

accggcaccg gcctgctgct gaccctgcag ccggaacaga aatttcagaa agtgaaaggc      240

tttggcggcg cgatgaccga tgcggcggcg ctgaacattc tggcgctgag cccgccggcg      300

cagaacctgc tgctgaaaag ctattttagc gaagaaggca ttggctataa cattattcgc      360

gtgccgatgg cgagctgcga ttttagcatt cgcacctata cctatgcgga taccccggat      420

gattttcagc tgcataactt tagcctgccg gaagaagata ccaaactgaa aattccgctg      480

attcatcgcg cgctgcagct ggcgcagcgc ccggtgagcc tgctggcgag cccgtggacc      540

agcccgacct ggctgaaaac caacggcgcg gtgaacggca aaggcagcct gaaaggccag      600

ccgggcgata tttatcatca gacctgggcg cgctattttg tgaaatttct ggatgcgtat      660

gcggaacata aactgcagtt ttgggcggtg accgcggaaa acgaaccgag cgcgggcctg      720

ctgagcggct atccgtttca gtgcctgggc tttaccccgg aacatcagcg cgattttatt      780

gcgcgcgatc tgggcccgac cctggcgaac agcacccatc ataacgtgcg cctgctgatg      840

ctggatgatc agcgcctgct gctgccgcat tgggcgaaag tggtgctgac cgatccggaa      900

gcggcgaaat atgtgcatgg cattgcggtg cattggtatc tggattttct ggcgccggcg      960

aaagcgaccc tgggcgaaac ccatcgcctg tttccgaaca ccatgctgtt tgcgagcgaa     1020

gcgtgcgtgg gcagcaaatt ttgggaacag agcgtgcgcc tgggcagctg ggatcgcggc     1080

atgcagtata gccatagcat tattgaagat ctgctgtatc atgtggtggg ctggaccgat     1140

tggaacctgg cgctgaaccc ggaaggcggc ccgaactggg tgcgcaactt tgtggatagc     1200

ccgattattg tggatattac caaagatacc ttttataaac agccgatgtt ttatcatctg     1260

ggccatttta gcaaatttat tccggaaggc agccagcgcg tgggcctggt ggcgagccag     1320

aaaaacgatc tggatgcggt ggcgctgatg catccggatg gcagcgcggt ggtggtggtg     1380

ctgaaccgca gcagcaaaga tgtgccgctg accattaaag atccggcggt gggctttctg     1440

gaaaccatta gcccgggcta tagcattcat acctatctgt ggcatcgcca gtaa           1494


<210>  4
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified -glucocerebrosidase  (GCase variant)

<400>  4

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn His Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asn Leu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Ile Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


His Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Leu Gln Leu Ala Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Ile Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Arg Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Glu His Asn 
    210                 215                 220                 


Leu Gln Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Leu Ser Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asp Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Thr Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Leu Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ile Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Leu Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Lys Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp His Arg 
                485                 490                 495     


Gln 
    


<210>  5
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified -glucocerebrosidase  (GCase variant)

<400>  5
gcgcgcccgt gcattccgaa aagctttggc tatagcagcg tggtgtgcgt gtgcaacgcg       60

acctattgcg atagctttga tccgccgacc tttccggcgc cgggcacctt tagccgctat      120

gaaagcaccc gcagcggccg ccgcatggaa ctgagcatgg gcccgattca ggcgaaccat      180

accggcaccg gcctgctgct gaccctgcag ccggaacaga aatttcagaa agtgaaaggc      240

tttggcggcg cgatgaccga tgcggcggcg ctgaacattc tggcgctgag cccgccggcg      300

cagaacctgc tgctgaaaag ctattttagc gaagaaggca ttggctataa cattattcgc      360

gtgccgatgg cgagctgcga ttttagcatt cgcacctata cctatgcgga taccccggat      420

gattttcagc tgcataactt tagcctgccg gaagaagata ccaaactgaa aattccgctg      480

attcatcgcg cgctgcagct ggcgcagcgc ccggtgagcc tgctggcgag cccgtggacc      540

agcccgacct ggctgaaaac caacggcgcg gtgaacggca aaggcagcct gaaaggccag      600

ccgggcgata tttatcatca gacctgggcg cgctattttg tgaaatttct ggatgcgtat      660

gcggaacata acctgcagtt ttgggcggtg accgcggaaa acgaaccgag cgcgggcctg      720

ctgagcggct atccgtttca gtgcctgggc tttaccccgg aacatcagcg cgattttatt      780

gcgcgcgatc tgggcccgac cctggcgaac agcacccatc ataacgtgcg cctgctgatg      840

ctggatgatc agcgcctgct gctgccgcat tgggcgaaag tggtgctgac cgatccggaa      900

gcggcgaaat atgtgcatgg cattgcggtg cattggtatc tggattttct ggcgccggcg      960

aaagcgaccc tgggcgaaac ccatcgcctg tttccgaaca ccatgctgtt tgcgagcgaa     1020

gcgtgcgtgg gcagcaaatt ttgggaacag agcgtgcgcc tgggcagctg ggatcgcggc     1080

atgcagtata gccatagcat tattgaagat ctgctgtatc atgtggtggg ctggaccgat     1140

tggaacctgg cgctgaaccc ggaaggcggc ccgaactggg tgcgcaactt tgtggatagc     1200

ccgattattg tggatattac caaagatacc ttttataaac agccgatgtt ttatcatctg     1260

ggccatttta gcaaatttat tccggaaggc agccagcgcg tgggcctggt ggcgagccag     1320

aaaaacgatc tggatgcggt ggcgctgatg catccggatg gcagcgcggt ggtggtggtg     1380

ctgaaccgca gcagcaaaga tgtgccgctg accattaaag atccggcggt gggctttctg     1440

gaaaccatta gcccgggcta tagcattcat acctatctgt ggcatcgcca gtaa           1494


<210>  6
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified -glucocerebrosidase  (GCase variant)

<400>  6

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn His Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asn Leu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Ile Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Leu Gln Leu Ala Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Arg Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Asn 
    210                 215                 220                 


Leu Gln Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Leu Ser Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asp Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Phe Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ile Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Leu Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Lys Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp His Arg 
                485                 490                 495     


Gln 
    


<210>  7
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified -glucocerebrosidase  (GCase variant)

<400>  7
gcgcgcccgt gcattccgaa aagctttggc tatagcagcg tggtgtgcgt gtgcaacgcg       60

acctattgcg atagctttga tccgccgacc tttccggcgc cgggcacctt tagccgctat      120

gaaagcaccc gcagcggccg ccgcatggaa ctgagcatgg gcccgattca ggcgaaccat      180

accggcaccg gcctgctgct gaccctgcag ccggaacaga aatttcagaa agtgaaaggc      240

tttggcggcg cgatgaccga tgcggcggcg ctgaacattc tggcgctgag cccgccggcg      300

caggatctgc tgctgaaaag ctattttagc gaagaaggca ttggctataa cattattcgc      360

gtgccgatgg cgagctgcga ttttagcatt cgcacctata cctatgcgga taccccggat      420

gattttcagc tgaaaaactt tagcctgccg gaagaagata ccaaactgaa aattccgctg      480

attcatcgcg cgcagcagct ggcgcagcgc ccggtgagcc tgctggcgag cccgtggacc      540

agcccgacct ggctgaaaac caacggcgcg gtgaacggca aaggcagcct gaaaggccag      600

ccgggcgata aatatcatca gacctgggcg cgctattttg tgaaatttct ggatgcgtat      660

gcgaaacata acctgacctt ttgggcggtg accgcggaaa acgaaccgag cgcgggcctg      720

attccgggct atccgtttca gtgcctgggc tttaccccgg aacatcagcg cgattttatt      780

gcgcgcgatc tgggcccgac cctggcgaac agcacccatc ataacgtgcg cctgctgatg      840

ctggatgatc agcgcctgct gctgccgcat tgggcgaaag tggtgctgac cgatccggaa      900

gcggcgaaat atgtgcatgg cattgcggtg cattggtatc tggattttct ggcgccggcg      960

aaagcgaccc tgggcgaaac ccatcgcctg tttccgaact atatgctgtt tgcgagcgaa     1020

gcgtgcgtgg gcagcaaatt ttgggaacag agcgtgcgcc tgggcagctg ggatcgcggc     1080

atgcagtata gccatagcat tattgaagat ctgaactatc atgtggtggg ctggaccgat     1140

tggaacctgg cgctgaaccc ggaaggcggc ccgaactggg tgcgcaactt tgtggatagc     1200

ccgattattg tggatattac caaagatacc ttttataaac agccgatgtt ttatcatctg     1260

ggccatttta gcaaatttat tccggaaggc agccagcgcg tgggcctggt ggcgagccag     1320

aaaaacgatc tggatgcggt ggcgctgatg catccggatg gcagcgcggt ggtggtggtg     1380

ctgaaccgca gcagcaaaga tgtgccgctg accatttggg atccggcggt gggctttctg     1440

gaaaccatta gcccgggcta tagcattcat acctatctgt ggcgccgcca gtaa           1494


<210>  8
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified -glucocerebrosidase  (GCase variant)

<400>  8

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn His Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asp Leu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Ile Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Gln Gln Leu Ala Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Arg Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Asn 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asp Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Tyr Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ile Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Leu Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  9
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  9
gcgcgcccgt gcattccgaa aagctttggc tatagcagcg tggtgtgcgt gtgcaacgcg       60

acctattgcg atagctttga tccgccgacc tttccggcgc cgggcacctt tagccgctat      120

gaaagcaccc gcagcggccg ccgcatggaa cgcagcatgg gcccgattca ggcgaaccat      180

accggcaccg gcctgctgct gaccctgcag ccggaacaga aatttcagaa agtgaaaggc      240

tttggcggcg cgatgaccga tgcggcggcg ctgaacattc tggcgctgag cccgccggcg      300

caggatctgc tgctgaaaag ctattttagc gaagaaggca ttggctataa cattattcgc      360

gtgccgatgg cgagctgcga ttttagcacc cgcacctata cctatgcgga taccccggat      420

gattttaaac tgaaaaactt tagcctgccg gaagaagata ccaaactgaa aattccgctg      480

attcatcgcg cgcagcagct gagccagcgc ccggtgagcc tgctggcgag cccgtggacc      540

agcccgacct ggctgaaaac caacggcgcg gtgaacggca aaggcagcct gaaaggccag      600

ccgggcgata aatatcatca gacctgggcg aactattttg tgaaatttct ggatgcgtat      660

gcgaaacata acctgacctt ttgggcggtg accgcggaaa acgaaccgag cgcgggcctg      720

attccgggct atccgtttca gtgcctgggc tttaccccgg aacatcagcg cgattttatt      780

gcgcgcaacc tgggcccgac cctggcgaac agcacccatc ataacgtgcg cctgctgatg      840

ctggatgatc agcgcctgct gctgccgcat tgggcgaaag tggtgctgac cgatccggaa      900

gcggcgcgct atgtgcatgg cattgcggtg cattggtatc tggattttct ggcgccggcg      960

aaagcgaccc tgggcgaaac ccatcgcctg tttccgaact atatgctgtt tgcgagcgaa     1020

gcgtgcgtgg gcagcaaatt ttgggaacag agcgtgcgcc tgggcagctg ggatcgcggc     1080

atgcagtata gccatagcat tattgaagat ctgaactatc atgtggtggg ctggaccgat     1140

tggaacctgg cgctgaaccc ggaaggcggc ccgaactggg tgcgcaactt tgtggatagc     1200

ccgattattg tggatattac caaagatacc ttttataaac agccgatgtt ttatcatctg     1260

ggccatttta gcaaatttat tccggaaggc agccagcgcg tgggcctggt ggcgagccag     1320

aaaaacgatc tggatgcggt ggcgctgatg catccggatg gcagcgcggt ggtggtggtg     1380

ctgaaccgca gcagcaaaga tgtgccgctg accatttggg atccggcggt gggctttctg     1440

gaaaccatta gcccgggcta tagcattcat acctatctgt ggcgccgcca gtaa           1494


<210>  10
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  10

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn His Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asp Leu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Thr Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Gln Gln Leu Ser Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Arg Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Asn 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asn Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Tyr Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ile Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Leu Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  11
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  11
gcgcgcccgt gcattccgaa aagctttggc tatagcagcg tggtgtgcgt gtgcaacgcg       60

acctattgcg atagctttga tccgccgacc tttccggcgc cgggcacctt tagccgctat      120

gaaagcaccc gcagcggccg ccgcatggaa ctgagcatgg gcccgattca ggcgaaccat      180

accggcaccg gcctgctgct gaccctgcag ccggaacaga aatttcagaa agtgaaaggc      240

tttggcggcg cgatgaccga tgcggcggcg ctgaacattc tggcgctgag cccgccggcg      300

caggatctgc tgctgaaaag ctattttagc gaagaaggca ttggctataa cattattcgc      360

gtgccgatgg cgagctgcga ttttagcacc cgcacctata cctatgcgga taccccggat      420

gattttcagc tgaaaaactt tagcctgccg gaagaagata ccaaactgaa aattccgctg      480

attcatcgcg cgcagcagct gagccagcgc ccggtgagcc tgctggcgag cccgtggacc      540

agcccgacct ggctgaaaac caacggcgcg gtgaacggca aaggcagcct gaaaggccag      600

ccgggcgata aatatcatca gacctgggcg aactattttg tgaaatttct ggatgcgtat      660

gcgaaacata acctgacctt ttgggcggtg accgcggaaa acgaaccgag cgcgggcctg      720

attccgggct atccgtttca gtgcctgggc tttaccccgg aacatcagcg cgattttatt      780

gcgcgcaacc tgggcccgac cctggcgaac agcacccatc ataacgtgcg cctgctgatg      840

ctggatgatc agcgcctgct gctgccgcat tgggcgaaag tggtgctgac cgatccggaa      900

gcggcgcgct atgtgcatgg cattgcggtg cattggtatc tggattttct ggcgccggcg      960

aaagcgaccc tgggcgaaac ccatcgcctg tttccgaact atatgctgtt tgcgagcgaa     1020

gcgtgcgtgg gcagcaaatt ttgggaacag agcgtgcgcc tgggcagctg ggatcgcggc     1080

atgcagtata gccatagcat tattgaagat ctgaactatc atgtggtggg ctggaccgat     1140

tggaacctgg cgctgaaccc ggaaggcggc ccgaactggg tgcgcaactt tgtggatagc     1200

ccgattattg tggatattac caaagatacc ttttataaac agccgatgtt ttatcatctg     1260

ggccatttta gcaaatttat tccggaaggc agccagcgcg tgggcctggt ggcgagccag     1320

aaaaacgatc tggatgcggt ggcgctgatg catccggatg gcagcgcggt ggtggtggtg     1380

ctgaaccgca gcagcaaaga tgtgccgctg accatttggg atccggcggt gggctttctg     1440

gaaaccatta gcccgggcta tagcattcat acctatctgt ggcgccgcca gtaa           1494


<210>  12
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  12

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn His Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asp Leu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Thr Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Gln Gln Leu Ser Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Asn Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Asn 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asn Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Arg Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Tyr Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ile Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Leu Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  13
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  NA sequence of optimized ecoli D7 Gcase

<400>  13
gcgcgcccgt gcattccgaa aagctttggc tatagcagcg tggtgtgcgt gtgcaacgcg       60

acctattgcg atagctttga tccgccgacc tttccggcgc cgggcacctt tagccgctat      120

gaaagcaccc gcagcggccg ccgcatggaa ctgagcatgg gcccgattca ggcgaactgg      180

accggcaccg gcctgctgct gaccctgcag ccggaacaga aatttcagaa agtgaaaggc      240

tttggcggcg cgatgaccga tgcggcggcg ctgaacattc tggcgctgag cccgccggcg      300

caggataacc tgctgaaaag ctattttagc gaagaaggca ttggctataa cattattcgc      360

gtgccgatgg cgagctgcga ttttagcacc cgcacctata cctatgcgga taccccggat      420

gattttcagc tgaaaaactt tagcctgccg gaagaagata ccaaactgaa aattccgctg      480

attcatcgcg cgcaggcgct gagccagcgc ccggtgagcc tgctggcgag cccgtggacc      540

agcccgacct ggctgaaaac caacggcgcg gtgaacggca aaggcagcct gaaaggccag      600

ccgggcgata aatatcatca gacctgggcg aactattttg tgaaatttct ggatgcgtat      660

gcgaaacata acctgacctt ttgggcggtg accgcggaaa acgaaccgag cgcgggcctg      720

attccgggct atccgtttca gtgcctgggc tttaccccgg aacatcagcg cgattttatt      780

gcgcgcaacc tgggcccgac cctggcgaac agcacccatc gcaacgtgcg cctgctgatg      840

ctggatgatc agcgcctgct gctgccgcat tgggcgaaag tggtgctgac cgatccggaa      900

gcggcgcgct atgtgcatgg cattgcggtg cattggtatc tggattttct ggcgccggcg      960

aaagcgaccc tgggcgaaac ccatcgcctg tttccggatt atatgctgtt tgcgagcgaa     1020

gcgtgcgtgg gcagcaaatt ttgggaacag agcgtgcgcc tgggcagctg ggatcgcggc     1080

atgcagtata gccatagcat tattgaagat ctgaactatc atgtggtggg ctggaccgat     1140

tggaacctgg cgctggatcc ggaaggcggc ccgaactggg tgaaaaactt tgtggatagc     1200

ccgattattg tggataccac caaagatacc ttttataaac agccgatgtt ttatcatatg     1260

ggccatttta gcaaatttat tccggaaggc agccagcgcg tgggcctggt ggcgagccag     1320

aaaaacgatc tggatgcggt ggcgctgatg catccggatg gcagcgcggt ggtggtggtg     1380

ctgaaccgca gcagcaaaga tgtgccgctg accatttggg atccggcggt gggctttctg     1440

gaaaccatta gcccgggcta tagcattcat acctatctgt ggcgccgcca gtaa           1494


<210>  14
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AA sequence of D7 Gcase

<400>  14

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn Trp Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asp Asn Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Thr Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Gln Ala Leu Ser Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Asn Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Asn 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asn Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His Arg Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Arg Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asp Tyr Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asp Pro Glu Gly Gly Pro Asn Trp Val Lys Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Thr Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Met Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  15
<211>  24
<212>  DNA
<213>  Artificial sequence

<220>
<223>  NA sequence of FLAG peptide

<400>  15
gactacaaag acgatgacga caag                                              24


<210>  16
<211>  8
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AA sequence of FLAG peptide

<400>  16

Asp Tyr Lys Asp Asp Asp Asp Lys 
1               5               


<210>  17
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  17
gcccgcccct gcatccctaa aagcttcggc tacagctcgg tggtgtgtgt ctgcaatgcc       60

acatactgtg actcctttga ccccccgacc tttcctgccc ccggtacctt cagccgctat      120

gagagtacac gcagtgggcg acggatggag ctgagtatgg ggcccatcca ggctaattgg      180

acgggcacag gcctgctact gaccctgcag ccagaacaga agttccagaa aatcaaggga      240

tttggagggg ccatgacaga tgctgctgct ctcaacatcc ttaagctgtc accccctgcc      300

caagagaacc tacttaaatc gtacttctct gaagaaggaa tcggatataa catcatccgg      360

gtacccatgg ccagctgtga cttctccacc cgcacctaca cctatgcaga cacccctgat      420

gatttccagt tgaagaactt cagcctccca gaggaagata ccaagctcaa gatacccctg      480

attcaccgag cccaggcctt gagccagcgt cccgtttcac tccttgccag cccctggaca      540

tcacccactt ggctcaagac caatggagcg atgaatggga aggggtcact caagggacag      600

cccggagaca agtaccacca gacctgggcc aactactttg tgaagttcct ggatgcctat      660

gctaagcacg gcttaacctt ctgggcagtg acagctgaaa atgagccttc tgctgggctg      720

atccccggat accccttcca gtgcctgggc ttcacccctg aacatcagcg agacttcatt      780

gcccgtaacc taggtcctac cctcgccaac agtactcacc acaatgtccg cctactcatg      840

ctggatgacc aacgcttgct gctgccccac tgggcaaagg tggtactgac agacccagaa      900

gcagctaaat atgttcatgg cattgctgta cattggtacc tggactttct ggctccagcc      960

aaagacaccc taggggagac acaccgcctg ttccccaaca agatgctctt tgcctcagag     1020

gcctgtaccg gctccaagtt ctgggagcag agtgtgcggc taggctcctg ggatcgaggg     1080

gagcagtacg cccacagcat catcgaggac ctcaactact gggtggtcgg ctggaccgac     1140

tggaaccttg ccctgaaccc cgaaggagga cccaattggg tgcgtaactt tgtcgacagt     1200

cccatcattg tagacgccac caaggacgag ttttacaaac agcccatgtt ctaccacatc     1260

ggccacttca gcaagttcat tcctgagggc tcccagagag tggggctggt tgccagtcag     1320

aagaacgacc tggacgcagt ggcactgatg aaccccgatg gctctgctgt tgtggtcgtg     1380

ctaaaccgct cctctaagga tgtgcctctt accatctggg atcctgctgt gggcttcatc     1440

gagacaatct cacctggcta ctccattcac acctacctgt ggcgtcgcca gtga           1494


<210>  18
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  18

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn Trp Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Ile Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Lys Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Glu Asn Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Thr Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Gln Ala Leu Ser Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Met Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Asn Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Gly 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asn Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Asp Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Lys Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Thr Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Glu Gln Tyr Ala His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr Trp Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ala Thr Lys Asp Glu Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Ile Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met Asn Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Ala Val Gly Phe Ile 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  19
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  19
gcccgcccct gcatccctaa aagcttcggc tacagctcgg tggtgtgtgt ctgcaatgcc       60

acatactgtg actcctttga ccccccgacc tttcctgccc ccggtacctt cagccgctat      120

gagagtacac gcagtgggcg acggatggag ctgagtatgg ggcccatcca ggctaattgg      180

acgggcacag gcctgctact gaccctgcag ccagaacaga agttccagaa aatcaaggga      240

tttggagggg ccatgacaga tgctgctgct ctcaacatcc ttaagctgtc accccctgcc      300

caagagaacc tacttaaatc gtacttctct gaagaaggaa tcggatataa catcatccgg      360

gtacccatgg ccagctgtga cttctccacc cgcacctaca cctatgcaga cacccctgat      420

gatttccagt tgaagaactt cagcctccca gaggaagata ccaagctcaa gatacccctg      480

attaagcgag cccaggcctt gagccagcgt cccgtttcac tccttgccag cccctggaca      540

gcccccactt ggctcaagac caatggagcg atgaatggga aggggtcact caagggacag      600

cccggagaca agtaccacca gacctgggcc aactactttg tgaagttcct ggatgcctat      660

gctaagcacg gcttaacctt ctgggcagtg acagctgaaa atgagccttc tgctgggctg      720

atccccggat accccttcca gtgcctgggc ttcacccctg aacatcagcg agacttcatt      780

gcccgtaacc taggtcctac cctcgccaac agtactcacc acaatgtccg cctactcatg      840

ctggatgacc aacgcttgct gctgccccac tgggcaaagg tggtactgag cgacccagaa      900

gcagctaaat atgttcatgg cattgctgta cattggtacc tggactttct ggctccagcc      960

aaagacaccc taggggagac acaccgcctg ttccccaaca agttcctctt tgcctcagag     1020

gcctgtaccg gctcccactt ctgggagcag agtgtgcggc taggctcctg ggatcgaggg     1080

gagcagtacg cccacagcat catcgaggac ctcaactact gggtggtcgg ctggaccgac     1140

tggaaccttg ccctggaccc cgaaggagga cccaattggg tgaagaactt tgtcgacagt     1200

cccatcattg tagacgccac caaggacgag ttttacaaac agcccatgtt ctaccacatc     1260

ggccacttca gcaagttcat tcctgagggc gcccagagag tggggctggt tgccagtcag     1320

aagaacgacc tggacgcagt ggcactgatg aaccccgatg gctctgctgt tgtggtcgtg     1380

ctaaaccgct ccgacaagga tgtgcctctt accatctggg atcctgacgt gggcttcatc     1440

gagacaatct cacctggcta ctccattcac acctacctgt ggcgtcgcca gtga           1494


<210>  20
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  20

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn Trp Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Ile Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Lys Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Glu Asn Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Thr Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile Lys Arg Ala Gln Ala Leu Ser Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ala Pro Thr Trp Leu Lys Thr Asn Gly Ala Met Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Asn Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Gly 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asn Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Ser Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Asp Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Lys Phe Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Thr Gly Ser His Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Glu Gln Tyr Ala His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr Trp Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asp Pro Glu Gly Gly Pro Asn Trp Val Lys Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ala Thr Lys Asp Glu Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Ile Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ala Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met Asn Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Asp Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Asp Val Gly Phe Ile 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  21
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  21
gcccgcccct gcatccctaa aagcttcggc tacagctcgg tggtgtgtgt ctgcaatgcc       60

acatactgtg actcctttga ccccccgacc tttcctgccc ccggtacctt cagccgctat      120

gagagtacac gcagtgggaa gcggatggag aggagtatgg ggcccatcca ggctaattgg      180

acgggcacag gcctgctact gaccctgcac ccagaacaga agttccagaa aatcaaggga      240

tttggagggg ccatgacaga tgctgctgct atcaacatcc ttaagctgtc accccctgcc      300

caagaggagc tacttaaatc gtacttctct gaagaaggaa tcgagtataa catcatccgg      360

gtacccatgg gcagctgtga cttctccacc cgcacctaca cctatgcaga cacccctaac      420

gatttccagt tgaagaactt cagcctccca gaggaagata ccaagctcaa gatacccctg      480

attaagcgag cccaggcctt gagccagcgt cccgtttcac tccttgccag cccctggaca      540

gcccccactt ggctcaagac caatggagcg atgaatggga aggggaccct caagggacag      600

cccggagaca agtaccacca gacctgggcc aactactttg tgaagttcct ggatgcctat      660

gctaagcacg gcttaacctt ctgggcagtg acagctgaaa atgagccttc tgctgggctg      720

atccccggat accccttcca gtgcctgggc ttcacccctg aacatcagcg agacttcatt      780

gcccgtaacc taggtcctac cctcgccaac agtactcacc acaatgtccg cctactcatg      840

ctggatgacc aacgcttgct gctgccccac tgggcaaagg tggtactgag cgacccagaa      900

gcagctaaat atgttcatgg cattgctgta cattggtacc tggactttct ggctccagcc      960

aaagacaccc taggggagac acaccgcctg ttccccgaca agttcctctt tgcctcagag     1020

gcctgtaccg gctcccactt ctgggagcag agtgtgcggc taggctcctg ggatcgaggg     1080

gagcagtacg cccacagcat catcgaggac ctcaactact gggtggtcgg ctggaccgac     1140

tggaaccttg ccctggaccc cgaaggagga cccaattggg tgaagaactt tgtcgacagt     1200

cccatcattg tagacgccac caaggacgag ttttacaaac agcccatgtt ctaccacatg     1260

ggccacttca gcaagttcat tcctgagggc gcccagagag tggggctgag cgccagtcag     1320

aagaacgacc tggacgcagt ggcactgatg aaccccgatg gctctgctgt tgtggtcgtg     1380

ctaaaccgct ccgacaagga tgtgcctctt accatctggg atcctgacgt gggcttcatc     1440

gagacaatct cacctggcta ctccattcac acctacctgt ggcgtcgcca gtga           1494


<210>  22
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  22

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Lys Arg 
        35                  40                  45              


Met Glu Arg Ser Met Gly Pro Ile Gln Ala Asn Trp Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu His Pro Glu Gln Lys Phe Gln Lys Ile Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Ile Asn Ile Leu Lys Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Glu Glu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Glu Tyr Asn Ile Ile Arg Val Pro Met Gly Ser Cys Asp Phe 
        115                 120                 125             


Ser Thr Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asn Asp Phe Gln Leu 
    130                 135                 140                 


Lys Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile Lys Arg Ala Gln Ala Leu Ser Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ala Pro Thr Trp Leu Lys Thr Asn Gly Ala Met Asn 
            180                 185                 190         


Gly Lys Gly Thr Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Asn Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Gly 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asn Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Ser Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Asp Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asp Lys Phe Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Thr Gly Ser His Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Glu Gln Tyr Ala His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr Trp Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asp Pro Glu Gly Gly Pro Asn Trp Val Lys Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ala Thr Lys Asp Glu Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Met Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ala Gln 
            420                 425                 430         


Arg Val Gly Leu Ser Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met Asn Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Asp Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Asp Val Gly Phe Ile 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  23
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  NA sequence of optimized human D7 Gcase

<400>  23
gcccgcccct gcatccctaa aagcttcggc tacagctcgg tggtgtgtgt ctgcaatgcc       60

acatactgtg actcctttga ccccccgacc tttcctgccc ccggtacctt cagccgctat      120

gagagtacac gcagtgggaa gcggatggag aggagtatgg ggcccatcca ggctaattgg      180

acgggcacag gcctgctact gaccctgcac ccagaacaga agttccagaa aatcaaggga      240

tttggagggg ccatgacaga tgctgctgct atcaacatcc ttaagctgtc accccctgcc      300

caagaggagc tacttaaatc gtacttctct gaagaaggaa tcgagtataa catcatccgg      360

gtacccatgg gcagctgtga cttctccacc cgcacctaca cctatgcaga cacccctaac      420

gatttccagt tgaagaactt cagcctccca gaggaagata ccaagctcaa gatacccctg      480

attaagcgag cccaggcctt gagccagcgt cccgtttcac tccttgccag cccctggaca      540

gcccccactt ggctcaagac caatggagcg atgaatggga aggggaccct caagggacag      600

cccggagaca agtaccacca gacctgggcc aactactttg tgaagttcct ggatgcctat      660

gctaagcacg gcttaacctt ctgggcagtg acagctgaaa atgagccttc tgctgggctg      720

atccccggat accccttcca gtgcctgggc ttcacccctg aacatcagcg agacttcatt      780

gcccgtaacc taggtcctac cctcgccaac agtactcacc acaatgtccg cctactcatg      840

ctggatgacc aacgcttgct gctgccccac tgggcaaagg tggtactgag cgacccagaa      900

gcagctaaat atgttcatgg cattgctgta cattggtacc tggactttct ggctccagcc      960

aaagacaccc taggggagac acaccgcctg ttccccgaca agttcctctt tgcctcagag     1020

gcctgtaccg gctcccactt ctgggagcag agtgtgcggc taggctcctg ggatcgaggg     1080

gagcagtacg cccacagcat catcgaggac ctcaactact gggtggtcgg ctggaccgac     1140

tggaaccttg ccctggaccc cgaaggagga cccaattggg tgaagaactt tgtcgacagt     1200

cccatcattg tagacgccac caaggacgag ttttacaaac agcccatgtt ctaccacatg     1260

ggccacttca gcaagttcat tcctgagggc gcccagagag tggggctgag cgccagtcag     1320

aagaacgacc tggacgcagt ggcactgatg aaccccgatg gctctgctgt tgtggtcgtg     1380

ctaaaccgct ccgacaagga tgtgcctctt accatctggg atcctgacgt gggcttcatc     1440

gagacaatct cacctggcta ctccattcac acctacctgt ggcgtcgcca gtga           1494


<210>  24
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  NA sequence of WT Gcase (human optimized codons)

<400>  24
gcccgcccct gcatccctaa aagcttcggc tacagctcgg tggtgtgtgt ctgcaatgcc       60

acatactgtg actcctttga ccccccgacc tttcctgccc ttggtacctt cagccgctat      120

gagagtacac gcagtgggcg acggatggag ctgagtatgg ggcccatcca ggctaatcac      180

acgggcacag gcctgctact gaccctgcag ccagaacaga agttccagaa agtgaaggga      240

tttggagggg ccatgacaga tgctgctgct ctcaacatcc ttgccctgtc accccctgcc      300

caaaatttgc tacttaaatc gtacttctct gaagaaggaa tcggatataa catcatccgg      360

gtacccatgg ccagctgtga cttctccatc cgcacctaca cctatgcaga cacccctgat      420

gatttccagt tgcacaactt cagcctccca gaggaagata ccaagctcaa gatacccctg      480

attcaccgag ccctgcagtt ggcccagcgt cccgtttcac tccttgccag cccctggaca      540

tcacccactt ggctcaagac caatggagcg gtgaatggga aggggtcact caagggacag      600

cccggagaca tctaccacca gacctgggcc agatactttg tgaagttcct ggatgcctat      660

gctgagcaca agttacagtt ctgggcagtg acagctgaaa atgagccttc tgctgggctg      720

ttgagtggat accccttcca gtgcctgggc ttcacccctg aacatcagcg agacttcatt      780

gcccgtgacc taggtcctac cctcgccaac agtactcacc acaatgtccg cctactcatg      840

ctggatgacc aacgcttgct gctgccccac tgggcaaagg tggtactgac agacccagaa      900

gcagctaaat atgttcatgg cattgctgta cattggtacc tggactttct ggctccagcc      960

aaagccaccc taggggagac acaccgcctg ttccccaaca ccatgctctt tgcctcagag     1020

gcctgtgtgg gctccaagtt ctgggagcag agtgtgcggc taggctcctg ggatcgaggg     1080

atgcagtaca gccacagcat catcacgaac ctcctgtacc atgtggtcgg ctggaccgac     1140

tggaaccttg ccctgaaccc cgaaggagga cccaattggg tgcgtaactt tgtcgacagt     1200

cccatcattg tagacatcac caaggacacg ttttacaaac agcccatgtt ctaccacctt     1260

ggccacttca gcaagttcat tcctgagggc tcccagagag tggggctggt tgccagtcag     1320

aagaacgacc tggacgcagt ggcactgatg catcccgatg gctctgctgt tgtggtcgtg     1380

ctaaaccgct cctctaagga tgtgcctctt accatcaagg atcctgctgt gggcttcctg     1440

gagacaatct cacctggcta ctccattcac acctacctgt ggcgtcgcca gtga           1494


<210>  25
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AA sequence of WT Gcase

<400>  25

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Leu Gly Thr Phe Ser Arg Tyr Glu Ser Thr Arg Ser Gly Arg Arg 
        35                  40                  45              


Met Glu Leu Ser Met Gly Pro Ile Gln Ala Asn His Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu Gln Pro Glu Gln Lys Phe Gln Lys Val Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Leu Asn Ile Leu Ala Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Asn Leu Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Gly Tyr Asn Ile Ile Arg Val Pro Met Ala Ser Cys Asp Phe 
        115                 120                 125             


Ser Ile Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Gln Leu 
    130                 135                 140                 


His Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile His Arg Ala Leu Gln Leu Ala Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ser Pro Thr Trp Leu Lys Thr Asn Gly Ala Val Asn 
            180                 185                 190         


Gly Lys Gly Ser Leu Lys Gly Gln Pro Gly Asp Ile Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Arg Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Glu His Lys 
    210                 215                 220                 


Leu Gln Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Leu Ser Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asp Leu Gly Pro Thr Leu Ala Asn Ser Thr 
            260                 265                 270         


His His Asn Val Arg Leu Leu Met Leu Asp Asp Gln Arg Leu Leu Leu 
        275                 280                 285             


Pro His Trp Ala Lys Val Val Leu Thr Asp Pro Glu Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Lys Ala Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asn Thr Met Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Val Gly Ser Lys Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Met Gln Tyr Ser His Ser Ile Ile 
        355                 360                 365             


Thr Asn Leu Leu Tyr His Val Val Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asn Pro Glu Gly Gly Pro Asn Trp Val Arg Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ile Thr Lys Asp Thr Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Leu Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ser Gln 
            420                 425                 430         


Arg Val Gly Leu Val Ala Ser Gln Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Met His Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Ser Lys Asp Val Pro Leu Thr Ile Lys Asp Pro Ala Val Gly Phe Leu 
465                 470                 475                 480 


Glu Thr Ile Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  26
<211>  1494
<212>  DNA
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  26
gcccgcccct gcatccctaa aagcttcggc tacagctcgg tggtgtgtgt ctgcaatgcc       60

acatactgtg actcctttga ccccccgacc tttcctgccc ccggtcagtt cgcccgctat      120

gagagtacac gcagtgggaa gcggatggag aggagtatgg ggcccatcca ggctaattgg      180

acgggcacag gcctgctact gaccctgcac ccagaacaga agttccagaa aatcaaggga      240

tttggagggg ccatgacaga tgctgctgct atcaacatcc ttaagctgtc accccctgcc      300

caagagaggc tacttaaatc gtacttctct gaagaaggaa tcgagtataa catcatccgg      360

gtacccatgg gcagctgtga cttctccacc cgcacctaca cctatgcaga cacccctgac      420

gatttcgagt tgagaaactt cagcctccca gaggaagata ccaagctcaa gatacccctg      480

attaagcgag cccaggcctt gagccagcgt cccgtttcac tccttgccag cccctggaca      540

gcccccgcct ggatgaagac caatggagcg atgaatggga aggggaccct caagggacag      600

cccggagaca agtaccacca gacctgggcc aactactttg tgaagttcct ggatgcctat      660

gctaagcacg gcttaacctt ctgggcagtg acagctgaaa atgagccttc tgctgggctg      720

atccccggat accccttcca gtgcctgggc ttcacccctg aacatcagcg agacttcatt      780

gcccgtaacc taggtcctac cctcgccaac agtagccaca aggacgtccg cctactcatg      840

ctggatgacc aacgcagcct gctgccccac tgggcacagg tggtactgag cgacccaaga      900

gcagctaaat atgttcatgg cattgctgta cattggtacc tggactttct ggctccagcc      960

gaggacaccc taggggagac acaccgcctg ttccccgaca agttcctctt tgcctcagag     1020

gcctgtaccg gctcccactt ctgggagcag agtgtgcggc taggctcctg ggatcgaggg     1080

gagcagtacg cccacagcat catcgaggac ctcaactact gggtgaccgg ctggaccgac     1140

tggaaccttg ccctggaccc cgaaggagga cccaattggg tgaagaactt tgtcgacagt     1200

cccatcattg tagacgccac cagggacgag ttttacaaac agcccatgtt ctaccacatg     1260

ggccacttca gcaagttcat tcctgagggc gcccagagag tggggctgag cgccagtgag     1320

aagaacgacc tggacgcagt ggcactgcag aaccccgatg gctctgctgt tgtggtcgtg     1380

ctaaaccgct ccgacaagga tgtgcctctt accatctggg atcctgacgt gggcttcatc     1440

gagacagtgt cacctggcta ctccattcac acctacctgt ggcgtcgcca gtga           1494


<210>  27
<211>  497
<212>  PRT
<213>  Artificial sequence

<220>
<223>  genetically modified betta-glucocerebrosidase  (GCase variant)

<400>  27

Ala Arg Pro Cys Ile Pro Lys Ser Phe Gly Tyr Ser Ser Val Val Cys 
1               5                   10                  15      


Val Cys Asn Ala Thr Tyr Cys Asp Ser Phe Asp Pro Pro Thr Phe Pro 
            20                  25                  30          


Ala Pro Gly Gln Phe Ala Arg Tyr Glu Ser Thr Arg Ser Gly Lys Arg 
        35                  40                  45              


Met Glu Arg Ser Met Gly Pro Ile Gln Ala Asn Trp Thr Gly Thr Gly 
    50                  55                  60                  


Leu Leu Leu Thr Leu His Pro Glu Gln Lys Phe Gln Lys Ile Lys Gly 
65                  70                  75                  80  


Phe Gly Gly Ala Met Thr Asp Ala Ala Ala Ile Asn Ile Leu Lys Leu 
                85                  90                  95      


Ser Pro Pro Ala Gln Glu Arg Leu Leu Lys Ser Tyr Phe Ser Glu Glu 
            100                 105                 110         


Gly Ile Glu Tyr Asn Ile Ile Arg Val Pro Met Gly Ser Cys Asp Phe 
        115                 120                 125             


Ser Thr Arg Thr Tyr Thr Tyr Ala Asp Thr Pro Asp Asp Phe Glu Leu 
    130                 135                 140                 


Arg Asn Phe Ser Leu Pro Glu Glu Asp Thr Lys Leu Lys Ile Pro Leu 
145                 150                 155                 160 


Ile Lys Arg Ala Gln Ala Leu Ser Gln Arg Pro Val Ser Leu Leu Ala 
                165                 170                 175     


Ser Pro Trp Thr Ala Pro Ala Trp Met Lys Thr Asn Gly Ala Met Asn 
            180                 185                 190         


Gly Lys Gly Thr Leu Lys Gly Gln Pro Gly Asp Lys Tyr His Gln Thr 
        195                 200                 205             


Trp Ala Asn Tyr Phe Val Lys Phe Leu Asp Ala Tyr Ala Lys His Gly 
    210                 215                 220                 


Leu Thr Phe Trp Ala Val Thr Ala Glu Asn Glu Pro Ser Ala Gly Leu 
225                 230                 235                 240 


Ile Pro Gly Tyr Pro Phe Gln Cys Leu Gly Phe Thr Pro Glu His Gln 
                245                 250                 255     


Arg Asp Phe Ile Ala Arg Asn Leu Gly Pro Thr Leu Ala Asn Ser Ser 
            260                 265                 270         


His Lys Asp Val Arg Leu Leu Met Leu Asp Asp Gln Arg Ser Leu Leu 
        275                 280                 285             


Pro His Trp Ala Gln Val Val Leu Ser Asp Pro Arg Ala Ala Lys Tyr 
    290                 295                 300                 


Val His Gly Ile Ala Val His Trp Tyr Leu Asp Phe Leu Ala Pro Ala 
305                 310                 315                 320 


Glu Asp Thr Leu Gly Glu Thr His Arg Leu Phe Pro Asp Lys Phe Leu 
                325                 330                 335     


Phe Ala Ser Glu Ala Cys Thr Gly Ser His Phe Trp Glu Gln Ser Val 
            340                 345                 350         


Arg Leu Gly Ser Trp Asp Arg Gly Glu Gln Tyr Ala His Ser Ile Ile 
        355                 360                 365             


Glu Asp Leu Asn Tyr Trp Val Thr Gly Trp Thr Asp Trp Asn Leu Ala 
    370                 375                 380                 


Leu Asp Pro Glu Gly Gly Pro Asn Trp Val Lys Asn Phe Val Asp Ser 
385                 390                 395                 400 


Pro Ile Ile Val Asp Ala Thr Arg Asp Glu Phe Tyr Lys Gln Pro Met 
                405                 410                 415     


Phe Tyr His Met Gly His Phe Ser Lys Phe Ile Pro Glu Gly Ala Gln 
            420                 425                 430         


Arg Val Gly Leu Ser Ala Ser Glu Lys Asn Asp Leu Asp Ala Val Ala 
        435                 440                 445             


Leu Gln Asn Pro Asp Gly Ser Ala Val Val Val Val Leu Asn Arg Ser 
    450                 455                 460                 


Asp Lys Asp Val Pro Leu Thr Ile Trp Asp Pro Asp Val Gly Phe Ile 
465                 470                 475                 480 


Glu Thr Val Ser Pro Gly Tyr Ser Ile His Thr Tyr Leu Trp Arg Arg 
                485                 490                 495     


Gln 
    


<210>  28
<211>  87
<212>  DNA
<213>  Artificial sequence

<220>
<223>  NA TwinStrep

<400>  28
tggtcccacc cccagttcga gaagggcggc ggtagtggag ggggcggatc tggcggctca       60

gcttggagcc acccccagtt cgaaaag                                           87


<210>  29
<211>  29
<212>  PRT
<213>  Artificial sequence

<220>
<223>  AA TwinStrep

<400>  29

Trp Ser His Pro Gln Phe Glu Lys Gly Gly Gly Ser Gly Gly Gly Gly 
1               5                   10                  15      


Ser Gly Gly Ser Ala Trp Ser His Pro Gln Phe Glu Lys 
            20                  25                  


