                         SEQUENCE LISTING

<110>  Waterford Institute of Technology
 
<120>  Isonitrile- and Nitrile-Metabolising Enzymes and a Process for Producing Amides 
       and Acids using Same

<130>  P120820.WO.01

<150>  GB111641.5
<151>  2011-07-07

<160>  14    

<170>  PatentIn version 3.5

<210>  1
<211>  1080
<212>  DNA
<213>  Arthrobacter sp. SS-24


<220>
<221>  CDS
<222>  (1)..(1080)

<400>  1
atg gac cat ccc aag ttc aag gca gcg gcc att cag gcc gca cct gtc         48
Met Asp His Pro Lys Phe Lys Ala Ala Ala Ile Gln Ala Ala Pro Val           
1               5                   10                  15                

ttc ctc aat ctt gac gcc acc atc gac aag gcg gtc gcc ctc ata gag         96
Phe Leu Asn Leu Asp Ala Thr Ile Asp Lys Ala Val Ala Leu Ile Glu           
            20                  25                  30                    

gag gcc tcc agc aat ggt gct gaa gtg ata gct ttc ccc gag acc tgg        144
Glu Ala Ser Ser Asn Gly Ala Glu Val Ile Ala Phe Pro Glu Thr Trp           
        35                  40                  45                        

ctt ccg ggc tac ccc tgg tac gcc tgg ctg gat gcg ccc gca cta tgg        192
Leu Pro Gly Tyr Pro Trp Tyr Ala Trp Leu Asp Ala Pro Ala Leu Trp           
    50                  55                  60                            

ctg gcg aaa ttc ggt cag cgc tac ttc gac aac tcc ctc gag tac ggt        240
Leu Ala Lys Phe Gly Gln Arg Tyr Phe Asp Asn Ser Leu Glu Tyr Gly           
65                  70                  75                  80            

act ccc cag gcc gaa cgt gtt gcc aac gct gcc aag gac aac aac atc        288
Thr Pro Gln Ala Glu Arg Val Ala Asn Ala Ala Lys Asp Asn Asn Ile           
                85                  90                  95                

atg gtc ggt atg ggc ctg agc gaa cga agc ggc agc agt ctc tac atc        336
Met Val Gly Met Gly Leu Ser Glu Arg Ser Gly Ser Ser Leu Tyr Ile           
            100                 105                 110                   

gcc caa tgg att atc ggc aac gac gga aaa acc atc acc caa cgc cgt        384
Ala Gln Trp Ile Ile Gly Asn Asp Gly Lys Thr Ile Thr Gln Arg Arg           
        115                 120                 125                       

aag ctc aag ccc acg cat gtg gaa cgt acc ata tat ggc gaa ggg gac        432
Lys Leu Lys Pro Thr His Val Glu Arg Thr Ile Tyr Gly Glu Gly Asp           
    130                 135                 140                           

ggc agc gat ctt tcc gtc tgg gac acg aac ttg ggg cgt gtg ggt ggc        480
Gly Ser Asp Leu Ser Val Trp Asp Thr Asn Leu Gly Arg Val Gly Gly           
145                 150                 155                 160           

ctt tgc tgc tgg gaa cac ctg cag ccg ctg tcc aag tac gcc atg tac        528
Leu Cys Cys Trp Glu His Leu Gln Pro Leu Ser Lys Tyr Ala Met Tyr           
                165                 170                 175               

gcg cag aac gaa caa gta cat ttc gcg gcc tgg cct agc ttc tcc ctc        576
Ala Gln Asn Glu Gln Val His Phe Ala Ala Trp Pro Ser Phe Ser Leu           
            180                 185                 190                   

tat gag ggc gga gca cat gca ctc agc gct gaa gcg aac gtt gca gct        624
Tyr Glu Gly Gly Ala His Ala Leu Ser Ala Glu Ala Asn Val Ala Ala           
        195                 200                 205                       

tca cag gtg tat gca ctg gaa gga tca tgc ttc gtg ctg gcg cct aca        672
Ser Gln Val Tyr Ala Leu Glu Gly Ser Cys Phe Val Leu Ala Pro Thr           
    210                 215                 220                           

gcc act gtt tcc caa gag atg cag gac gag ttg tgc gag acg gac ctg        720
Ala Thr Val Ser Gln Glu Met Gln Asp Glu Leu Cys Glu Thr Asp Leu           
225                 230                 235                 240           

cag agg gct ctc ctg aaa act ggt ggt ggg tac agc cgt atc ttc ggc        768
Gln Arg Ala Leu Leu Lys Thr Gly Gly Gly Tyr Ser Arg Ile Phe Gly           
                245                 250                 255               

ccg gat ggc aaa cag ctc aac gaa tcc ctt ccg acg gaa gag gaa ggg        816
Pro Asp Gly Lys Gln Leu Asn Glu Ser Leu Pro Thr Glu Glu Glu Gly           
            260                 265                 270                   

att gtc tac gca cac atc gat ctt gga aaa atc tcc ctt tcc aag gcc        864
Ile Val Tyr Ala His Ile Asp Leu Gly Lys Ile Ser Leu Ser Lys Ala           
        275                 280                 285                       

atc gca gac ccg gcc ggt cac tac tca cga gcg gac gcc acc caa ctt        912
Ile Ala Asp Pro Ala Gly His Tyr Ser Arg Ala Asp Ala Thr Gln Leu           
    290                 295                 300                           

gtc ctc aac aag acc gtc cgc cgc ccg gtc atc gaa acc tca ctg gtc        960
Val Leu Asn Lys Thr Val Arg Arg Pro Val Ile Glu Thr Ser Leu Val           
305                 310                 315                 320           

gag caa gca act cgc acc cct gcc ggg aat tcc atg ccg ggc cct gcc       1008
Glu Gln Ala Thr Arg Thr Pro Ala Gly Asn Ser Met Pro Gly Pro Ala           
                325                 330                 335               

cct gca gac cgg tcc tgg gat gct gca tcc gcg gaa atg gaa gga aac       1056
Pro Ala Asp Arg Ser Trp Asp Ala Ala Ser Ala Glu Met Glu Gly Asn           
            340                 345                 350                   

gag cag gtt cca cag act ctc tag                                       1080
Glu Gln Val Pro Gln Thr Leu                                               
        355                                                               


<210>  2
<211>  359
<212>  PRT
<213>  Arthrobacter sp. SS-24

<400>  2

Met Asp His Pro Lys Phe Lys Ala Ala Ala Ile Gln Ala Ala Pro Val 
1               5                   10                  15      


Phe Leu Asn Leu Asp Ala Thr Ile Asp Lys Ala Val Ala Leu Ile Glu 
            20                  25                  30          


Glu Ala Ser Ser Asn Gly Ala Glu Val Ile Ala Phe Pro Glu Thr Trp 
        35                  40                  45              


Leu Pro Gly Tyr Pro Trp Tyr Ala Trp Leu Asp Ala Pro Ala Leu Trp 
    50                  55                  60                  


Leu Ala Lys Phe Gly Gln Arg Tyr Phe Asp Asn Ser Leu Glu Tyr Gly 
65                  70                  75                  80  


Thr Pro Gln Ala Glu Arg Val Ala Asn Ala Ala Lys Asp Asn Asn Ile 
                85                  90                  95      


Met Val Gly Met Gly Leu Ser Glu Arg Ser Gly Ser Ser Leu Tyr Ile 
            100                 105                 110         


Ala Gln Trp Ile Ile Gly Asn Asp Gly Lys Thr Ile Thr Gln Arg Arg 
        115                 120                 125             


Lys Leu Lys Pro Thr His Val Glu Arg Thr Ile Tyr Gly Glu Gly Asp 
    130                 135                 140                 


Gly Ser Asp Leu Ser Val Trp Asp Thr Asn Leu Gly Arg Val Gly Gly 
145                 150                 155                 160 


Leu Cys Cys Trp Glu His Leu Gln Pro Leu Ser Lys Tyr Ala Met Tyr 
                165                 170                 175     


Ala Gln Asn Glu Gln Val His Phe Ala Ala Trp Pro Ser Phe Ser Leu 
            180                 185                 190         


Tyr Glu Gly Gly Ala His Ala Leu Ser Ala Glu Ala Asn Val Ala Ala 
        195                 200                 205             


Ser Gln Val Tyr Ala Leu Glu Gly Ser Cys Phe Val Leu Ala Pro Thr 
    210                 215                 220                 


Ala Thr Val Ser Gln Glu Met Gln Asp Glu Leu Cys Glu Thr Asp Leu 
225                 230                 235                 240 


Gln Arg Ala Leu Leu Lys Thr Gly Gly Gly Tyr Ser Arg Ile Phe Gly 
                245                 250                 255     


Pro Asp Gly Lys Gln Leu Asn Glu Ser Leu Pro Thr Glu Glu Glu Gly 
            260                 265                 270         


Ile Val Tyr Ala His Ile Asp Leu Gly Lys Ile Ser Leu Ser Lys Ala 
        275                 280                 285             


Ile Ala Asp Pro Ala Gly His Tyr Ser Arg Ala Asp Ala Thr Gln Leu 
    290                 295                 300                 


Val Leu Asn Lys Thr Val Arg Arg Pro Val Ile Glu Thr Ser Leu Val 
305                 310                 315                 320 


Glu Gln Ala Thr Arg Thr Pro Ala Gly Asn Ser Met Pro Gly Pro Ala 
                325                 330                 335     


Pro Ala Asp Arg Ser Trp Asp Ala Ala Ser Ala Glu Met Glu Gly Asn 
            340                 345                 350         


Glu Gln Val Pro Gln Thr Leu 
        355                 


<210>  3
<211>  359
<212>  PRT
<213>  Arthrobacter sp. SS-24

<400>  3

Met Asp His Pro Lys Phe Lys Ala Ala Ala Ile Gln Ala Ala Pro Val 
1               5                   10                  15      


Phe Leu Asn Leu Asp Ala Thr Ile Asp Lys Ala Val Ala Leu Ile Glu 
            20                  25                  30          


Glu Ala Ser Ser Asn Gly Ala Glu Val Ile Ala Phe Pro Glu Thr Trp 
        35                  40                  45              


Leu Pro Gly Tyr Pro Trp Tyr Ala Trp Leu Asp Ala Pro Ala Leu Trp 
    50                  55                  60                  


Leu Ala Lys Phe Gly Gln Arg Tyr Phe Asp Asn Ser Leu Glu Tyr Gly 
65                  70                  75                  80  


Thr Pro Gln Ala Glu Arg Val Ala Asn Ala Ala Lys Asp Asn Asn Ile 
                85                  90                  95      


Met Val Gly Met Gly Leu Ser Glu Arg Ser Gly Ser Ser Leu Tyr Ile 
            100                 105                 110         


Ala Gln Trp Ile Ile Gly Asn Asp Gly Lys Thr Ile Thr Gln Arg Arg 
        115                 120                 125             


Lys Leu Lys Pro Thr His Val Glu Arg Thr Ile Tyr Gly Glu Gly Asp 
    130                 135                 140                 


Gly Ser Asp Leu Ser Val Trp Asp Thr Asn Leu Gly Arg Val Gly Gly 
145                 150                 155                 160 


Leu Cys Cys Trp Glu His Leu Gln Pro Leu Ser Lys Tyr Ala Met Tyr 
                165                 170                 175     


Ala Gln Asn Glu Gln Val His Phe Ala Ala Trp Pro Ser Phe Ser Leu 
            180                 185                 190         


Tyr Glu Gly Gly Ala His Ala Leu Ser Ala Glu Ala Asn Val Ala Ala 
        195                 200                 205             


Ser Gln Val Tyr Ala Leu Glu Gly Ser Cys Phe Val Leu Ala Pro Thr 
    210                 215                 220                 


Ala Thr Val Ser Gln Glu Met Gln Asp Glu Leu Cys Glu Thr Asp Leu 
225                 230                 235                 240 


Gln Arg Ala Leu Leu Lys Thr Gly Gly Gly Tyr Ser Arg Ile Phe Gly 
                245                 250                 255     


Pro Asp Gly Lys Gln Leu Asn Glu Ser Leu Pro Thr Glu Glu Glu Gly 
            260                 265                 270         


Ile Val Tyr Ala His Ile Asp Leu Gly Lys Ile Ser Leu Ser Lys Ala 
        275                 280                 285             


Ile Ala Asp Pro Ala Gly His Tyr Ser Arg Ala Asp Ala Thr Gln Leu 
    290                 295                 300                 


Val Leu Asn Lys Thr Val Arg Arg Pro Val Ile Glu Thr Ser Leu Val 
305                 310                 315                 320 


Glu Gln Ala Thr Arg Thr Pro Ala Gly Asn Ser Met Pro Gly Pro Ala 
                325                 330                 335     


Pro Ala Asp Arg Ser Trp Asp Ala Ala Ser Ala Glu Met Glu Gly Asn 
            340                 345                 350         


Glu Gln Val Pro Gln Thr Leu 
        355                 


<210>  4
<211>  1246
<212>  DNA
<213>  Arthrobacter sp. SS-24

<400>  4
ggcgaacggg agagtaacac gtgagtaacc tgcccttgac tctgggataa gcctgggaaa       60

ctgggtctaa taccggatac gaccatctgg cgcatgtcat ggtggtggaa agcttttgtg      120

gttttggatg gactcgcggc ctatcagctt gttggtgggg taatggccta ccaaggcgac      180

gacgggtagc cggcctgaga gggtgaccgg ccacactggg actgagacac ggcccagact      240

cctacgggag gcagcagtgg ggaatattgc acaatgggcg caagcctgat gcagcgacgc      300

cgcgtgaggg atgacggcct tcgggttgta aacctctttc agtagggaag aagcgaaagt      360

gacggtacct gcagaagaag cgccggctaa ctacgtgcca gcagccgcgg taatacgtag      420

ggcgcaagcg ttatccggaa ttattgggcg taaagagctc gtaggcggtt tgtcgcgtct      480

gctgtgaaag accggggctc aactccggtt ctgcagtggg tacgggcaga ctagagtgca      540

gtaggggaga ctggaattcc tggtgtagcg gtgaaatgcg cagatatcag gaggaacacc      600

gatggcgaag gcaggtctct gggctgtaac tgacgctgag gagcgaaagc atggggagcg      660

aacaggatta gataccctgg tagtccatgc cgtaaacgtt gggcactagg tgtgggggac      720

attccacgtt ttccgcgccg tagctaacgc attaagtgcc ccgcctgggg agtacggccg      780

caaggctaaa actcaaagga attgacgggg gcccgcacaa gcggcggagc atgcggatta      840

attcgatgca acgcgaagaa ccttaccaag gcttgacatg aaccggaaag acctggaaac      900

aggtgccccg cttgcggtcg gtttacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg      960

agatgttggg ttaagtcccg caacgagcgc aaccctcgtt ctatgttgcc agcgcgttat     1020

ggcggggact cataggagac tgccggggtc aactcggagg aaggtgggga cgacgtcaaa     1080

tcatcatgcc ccttatgtct tgggcttcac gcatgctaca atggccggta caaagggttg     1140

cgatactgtg aggtggagct aatcccaaaa agccggtctc agttcggatt ggggtctgca     1200

actcgacccc atgaagtcgg agtcgctagt aatcgcagat cagcaa                    1246


<210>  5
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  1B F primer

<400>  5
tgytgggarc aytacaaycc                                                   20


<210>  6
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  1B R primer

<400>  6
tccatcatbc kyttkcgytt                                                   20


<210>  7
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  2A F primer

<400>  7
gsvytbtgct gytgggarca                                                   20


<210>  8
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  2A R1 primer

<400>  8
rtartgvccr gcvggrtc                                                     18


<210>  9
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  2A R2 primer

<400>  9
gartartgsc cgrcsggrtc                                                   20


<210>  10
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3A F primer

<400>  10
caycgcaarc tscarccsac                                                   20


<210>  11
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ISO TF primer

<400>  11
atggaccatc ccaagttcaa g                                                 21


<210>  12
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ISO TR primer

<400>  12
ctagagagtc tgtggaacct                                                   20


<210>  13
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ISO PF primer

<400>  13
gacgacgaca agatggacca tcccaagttc aag                                    33


<210>  14
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ISO PR primer

<400>  14
gaggagaagc ccggtctaga gagtctgtgg aac                                    33


