                         SEQUENCE LISTING

<110>  The Dow Chemical Company
       Sanghani, Paresh
       Delaplane, Sarah
 
<120>  Processes to Prepare Elongated 2-Ketoacids and C6-C10 Compounds 
       Therefrom via Genetic Modifications to Microbial Metabolic 
       Pathways

<130>  DOW 79045 MA / 40224.368

<160>  8     

<170>  PatentIn version 3.5

<210>  1
<211>  523
<212>  PRT
<213>  Citrobacter freundii

<400>  1

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Gln 
    50                  55                  60                  


Val Lys Asn Ser Arg Val Cys Ala Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


His Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Ser Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Thr Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Asn Ile Pro Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Gly Ala Ile Ile Ser Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Gly Val Gly Asn Ala Leu Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Leu Asn 
                245                 250                 255     


Val Gln Thr Arg Ile Asn His Gln Glu Ile Trp Arg Thr Ser Gln Leu 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Ser Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Asp Glu Met Gly Tyr Lys Glu Asn Glu Tyr Ser 
            340                 345                 350         


Leu Asp Asn Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Asn Asp Ile Ala Thr Ala Ser Val Lys Leu Ala Cys Gly Glu 
                405                 410                 415     


Glu Val Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asn Ile Glu Leu Val Lys 
        435                 440                 445             


Tyr Ser Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Gly Gln Val 
    450                 455                 460                 


Asp Ile Val Ala Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Glu Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Asn Glu Asn Asn Lys Glu Thr Val 
        515                 520             


<210>  2
<211>  523
<212>  PRT
<213>  Enterobacter cloacae

<400>  2

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Thr 
    50                  55                  60                  


Ile Lys Asn Ser Arg Val Cys Gly Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


His Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Ser Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Asp Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Asn Ile Pro Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Ser Asn Ile Ile Thr Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Ala Val Gly Asn Ala Ile Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Met Asn 
                245                 250                 255     


Val His Thr Arg Ile Asn His Asn Glu Ile Trp Arg Thr Ser Gln Thr 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Thr Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Glu Glu Met Gly Tyr Lys Asp Ser Asp Tyr Asn 
            340                 345                 350         


Met Asp Gln Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Ser Asp Ile Ala Thr Ala Ser Ile Lys Leu Ala Cys Gly Asp 
                405                 410                 415     


Glu Ile Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asp Val Glu Leu Val Lys 
        435                 440                 445             


Tyr Asp Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Gly Gln Val 
    450                 455                 460                 


Asp Ile Val Val Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Ala Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Lys Glu Asn Asn Lys Glu Thr Val 
        515                 520             


<210>  3
<211>  523
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthesized - Modified C. fruendii isopropylmalate synthase (H97A
       S139G N167G P169A G181A A182G G210A A214S G462D)

<400>  3

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Gln 
    50                  55                  60                  


Val Lys Asn Ser Arg Val Cys Ala Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


Ala Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Gly Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Thr Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Gly Ile Ala Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Ala Gly Ile Ile Ser Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Ala Val Gly Asn Ser Leu Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Leu Asn 
                245                 250                 255     


Val Gln Thr Arg Ile Asn His Gln Glu Ile Trp Arg Thr Ser Gln Leu 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Ser Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Asp Glu Met Gly Tyr Lys Glu Asn Glu Tyr Ser 
            340                 345                 350         


Leu Asp Asn Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Asn Asp Ile Ala Thr Ala Ser Val Lys Leu Ala Cys Gly Glu 
                405                 410                 415     


Glu Val Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asn Ile Glu Leu Val Lys 
        435                 440                 445             


Tyr Ser Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Asp Gln Val 
    450                 455                 460                 


Asp Ile Val Ala Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Glu Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Asn Glu Asn Asn Lys Glu Thr Val 
        515                 520             


<210>  4
<211>  523
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthesized - Modified C. freundii isopropylmalate (H97A S139G 
       N167G P169A G181A A182G G210A A214S Q258H R260A G462D)

<400>  4

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Gln 
    50                  55                  60                  


Val Lys Asn Ser Arg Val Cys Ala Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


Ala Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Gly Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Thr Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Gly Ile Ala Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Ala Gly Ile Ile Ser Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Ala Val Gly Asn Ser Leu Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Leu Asn 
                245                 250                 255     


Val His Thr Ala Ile Asn His Gln Glu Ile Trp Arg Thr Ser Gln Leu 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Ser Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Asp Glu Met Gly Tyr Lys Glu Asn Glu Tyr Ser 
            340                 345                 350         


Leu Asp Asn Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Asn Asp Ile Ala Thr Ala Ser Val Lys Leu Ala Cys Gly Glu 
                405                 410                 415     


Glu Val Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asn Ile Glu Leu Val Lys 
        435                 440                 445             


Tyr Ser Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Asp Gln Val 
    450                 455                 460                 


Asp Ile Val Ala Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Glu Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Asn Glu Asn Asn Lys Glu Thr Val 
        515                 520             


<210>  5
<211>  523
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthesized - Modified E. cloacae isopropylmalate synthase (H97A 
       S139G N167G P169A R260A N264Q G462D)

<400>  5

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Thr 
    50                  55                  60                  


Ile Lys Asn Ser Arg Val Cys Gly Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


Ala Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Gly Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Asp Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Gly Ile Ala Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Ser Asn Ile Ile Thr Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Ala Val Gly Asn Ala Ile Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Met Asn 
                245                 250                 255     


Val His Thr Ala Ile Asn His Gln Glu Ile Trp Arg Thr Ser Gln Thr 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Thr Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Glu Glu Met Gly Tyr Lys Asp Ser Asp Tyr Asn 
            340                 345                 350         


Met Asp Gln Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Ser Asp Ile Ala Thr Ala Ser Ile Lys Leu Ala Cys Gly Asp 
                405                 410                 415     


Glu Ile Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asp Val Glu Leu Val Lys 
        435                 440                 445             


Tyr Asp Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Asp Gln Val 
    450                 455                 460                 


Asp Ile Val Val Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Ala Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Lys Glu Asn Asn Lys Glu Thr Val 
        515                 520             


<210>  6
<211>  523
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthesized - Modified E. cloacae isopropylmalate synthase (H97A 
       S139G N167G P169A R260A N264Q G462D M255L)

<400>  6

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Thr 
    50                  55                  60                  


Ile Lys Asn Ser Arg Val Cys Gly Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


Ala Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Gly Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Asp Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Gly Ile Ala Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Ser Asn Ile Ile Thr Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Ala Val Gly Asn Ala Ile Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Leu Asn 
                245                 250                 255     


Val His Thr Ala Ile Asn His Gln Glu Ile Trp Arg Thr Ser Gln Thr 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Thr Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Glu Glu Met Gly Tyr Lys Asp Ser Asp Tyr Asn 
            340                 345                 350         


Met Asp Gln Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Ser Asp Ile Ala Thr Ala Ser Ile Lys Leu Ala Cys Gly Asp 
                405                 410                 415     


Glu Ile Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asp Val Glu Leu Val Lys 
        435                 440                 445             


Tyr Asp Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Asp Gln Val 
    450                 455                 460                 


Asp Ile Val Val Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Ala Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Lys Glu Asn Asn Lys Glu Thr Val 
        515                 520             


<210>  7
<211>  523
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthesized - Modified E. cloacase isopropylmalate synthase (H97A
       S139G N167G P169A D348E D350E M353L Q355N G462D)

<400>  7

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Thr 
    50                  55                  60                  


Ile Lys Asn Ser Arg Val Cys Gly Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


Ala Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Gly Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Asp Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Gly Ile Ala Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Ser Asn Ile Ile Thr Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Ala Val Gly Asn Ala Ile Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Met Asn 
                245                 250                 255     


Val His Thr Arg Ile Asn His Asn Glu Ile Trp Arg Thr Ser Gln Thr 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Thr Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Glu Glu Met Gly Tyr Lys Glu Ser Glu Tyr Asn 
            340                 345                 350         


Leu Asp Asn Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Ser Asp Ile Ala Thr Ala Ser Ile Lys Leu Ala Cys Gly Asp 
                405                 410                 415     


Glu Ile Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asp Val Glu Leu Val Lys 
        435                 440                 445             


Tyr Asp Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Asp Gln Val 
    450                 455                 460                 


Asp Ile Val Val Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Ala Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Lys Glu Asn Asn Lys Glu Thr Val 
        515                 520             


<210>  8
<211>  523
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthesized - Modified E. cloacae isopropylmalate synthase (H97A 
       S139G N167G P169A D348E D350E M353L Q355N G462D M255L R260A 
       N264Q)

<400>  8

Met Ser Gln Gln Val Ile Ile Phe Asp Thr Thr Leu Arg Asp Gly Glu 
1               5                   10                  15      


Gln Ala Leu Gln Ala Ser Leu Ser Val Lys Glu Lys Leu Gln Ile Ala 
            20                  25                  30          


Leu Ala Leu Glu Arg Met Gly Val Asp Val Met Glu Val Gly Phe Pro 
        35                  40                  45              


Val Ser Ser Pro Gly Asp Phe Glu Ser Val Gln Thr Ile Ala Arg Thr 
    50                  55                  60                  


Ile Lys Asn Ser Arg Val Cys Gly Leu Ala Arg Cys Val Glu Lys Asp 
65                  70                  75                  80  


Ile Asp Val Ala Ala Glu Ser Leu Lys Val Ala Glu Ala Phe Arg Ile 
                85                  90                  95      


Ala Thr Phe Ile Ala Thr Ser Pro Met His Ile Ala Thr Lys Leu Arg 
            100                 105                 110         


Ser Thr Leu Asp Glu Val Ile Glu Arg Ala Val Tyr Met Val Lys Arg 
        115                 120                 125             


Ala Arg Asn Tyr Thr Asp Asp Val Glu Phe Gly Cys Glu Asp Ala Gly 
    130                 135                 140                 


Arg Thr Pro Ile Asp Asp Leu Ala Arg Val Val Glu Ala Ala Ile Asn 
145                 150                 155                 160 


Ala Gly Ala Lys Thr Ile Gly Ile Ala Asp Thr Val Gly Tyr Thr Met 
                165                 170                 175     


Pro Phe Glu Phe Ser Asn Ile Ile Thr Gly Leu Tyr Glu Arg Val Pro 
            180                 185                 190         


Asn Ile Asp Lys Ala Ile Ile Ser Val His Thr His Asp Asp Leu Gly 
        195                 200                 205             


Leu Ala Val Gly Asn Ala Ile Ala Ala Val His Ala Gly Ala Arg Gln 
    210                 215                 220                 


Val Glu Gly Ala Met Asn Gly Ile Gly Glu Arg Ala Gly Asn Cys Ser 
225                 230                 235                 240 


Leu Glu Glu Val Ile Met Ala Ile Lys Val Arg Lys Asp Ile Leu Asn 
                245                 250                 255     


Val His Thr Ala Ile Asn His Gln Glu Ile Trp Arg Thr Ser Gln Thr 
            260                 265                 270         


Val Ser Gln Ile Cys Asn Met Pro Ile Pro Ala Asn Lys Ala Ile Val 
        275                 280                 285             


Gly Thr Gly Ala Phe Ala His Ser Ser Gly Ile His Gln Asp Gly Val 
    290                 295                 300                 


Leu Lys Asn Arg Glu Asn Tyr Glu Ile Met Thr Pro Glu Ser Ile Gly 
305                 310                 315                 320 


Leu Asn Gln Val Gln Leu Asn Leu Thr Ser Arg Ser Gly Arg Ala Ala 
                325                 330                 335     


Val Lys His Arg Met Glu Glu Met Gly Tyr Lys Glu Ser Glu Tyr Asn 
            340                 345                 350         


Leu Asp Asn Leu Tyr Asp Ala Phe Leu Lys Leu Ala Asp Lys Lys Gly 
        355                 360                 365             


Gln Val Phe Asp Tyr Asp Leu Glu Ala Leu Ala Phe Ile Asn Lys Gln 
    370                 375                 380                 


Gln Glu Glu Pro Glu His Phe Arg Leu Asp Tyr Phe Ser Val Gln Ser 
385                 390                 395                 400 


Gly Ser Ser Asp Ile Ala Thr Ala Ser Ile Lys Leu Ala Cys Gly Asp 
                405                 410                 415     


Glu Ile Lys Ala Glu Ala Ala Asn Gly Asn Gly Pro Val Asp Ala Ile 
            420                 425                 430         


Tyr Gln Ala Ile Asn Arg Val Thr Glu Tyr Asp Val Glu Leu Val Lys 
        435                 440                 445             


Tyr Asp Leu Thr Ala Lys Gly His Gly Lys Asp Ala Leu Asp Gln Val 
    450                 455                 460                 


Asp Ile Val Val Asn Tyr Asn Gly Arg Arg Phe His Gly Val Gly Leu 
465                 470                 475                 480 


Ala Thr Asp Ile Val Glu Ser Ser Ala Lys Ala Met Val His Val Leu 
                485                 490                 495     


Asn Asn Ile Trp Arg Ala Ala Glu Val Glu Lys Glu Leu Gln Arg Lys 
            500                 505                 510         


Ala Gln Asn Lys Glu Asn Asn Lys Glu Thr Val 
        515                 520             


