                         SEQUENCE LISTING

<110>  Texas A&M University System
       Raushel, Frank M.
       Bigley, Andrew N.
 
<120>  Variants of Phosphotriesterase for the Hydrolysis and 
       Detoxification of Nerve Agents

<130>  20155.0002.PZUS00

<160>  6     

<170>  PatentIn version 3.5

<210>  1
<211>  337
<212>  PRT
<213>  Brevundimonas diminuta

<400>  1

Met Ser Ile Gly Thr Gly Asp Arg Ile Asn Thr Val Arg Gly Pro Ile 
1               5                   10                  15      


Thr Ile Ser Glu Ala Gly Phe Thr Leu Thr His Glu His Ile Cys Gly 
            20                  25                  30          


Ser Ser Ala Gly Phe Leu Arg Ala Trp Pro Glu Phe Phe Gly Ser Arg 
        35                  40                  45              


Lys Ala Leu Ala Glu Lys Ala Val Arg Gly Leu Arg Arg Ala Arg Ala 
    50                  55                  60                  


Ala Gly Val Arg Thr Ile Val Asp Val Ser Thr Phe Asp Ile Gly Arg 
65                  70                  75                  80  


Asp Val Ser Leu Leu Ala Glu Val Ser Arg Ala Ala Asp Val His Ile 
                85                  90                  95      


Val Ala Ala Thr Gly Leu Trp Phe Asp Pro Pro Leu Ser Met Arg Leu 
            100                 105                 110         


Arg Ser Val Glu Glu Leu Thr Gln Phe Phe Leu Arg Glu Ile Gln Tyr 
        115                 120                 125             


Gly Ile Glu Asp Thr Gly Ile Arg Ala Gly Ile Ile Lys Val Ala Thr 
    130                 135                 140                 


Thr Gly Lys Ala Thr Pro Phe Gln Glu Leu Val Leu Lys Ala Ala Ala 
145                 150                 155                 160 


Arg Ala Ser Leu Ala Thr Gly Val Pro Val Thr Thr His Thr Ala Ala 
                165                 170                 175     


Ser Gln Arg Asp Gly Glu Gln Gln Ala Ala Ile Phe Glu Ser Glu Gly 
            180                 185                 190         


Leu Ser Pro Ser Arg Val Cys Ile Gly His Ser Asp Asp Thr Asp Asp 
        195                 200                 205             


Leu Ser Tyr Leu Thr Ala Leu Ala Ala Arg Gly Tyr Leu Ile Gly Leu 
    210                 215                 220                 


Asp His Ile Pro His Ser Ala Ile Gly Leu Glu Asp Asn Ala Ser Ala 
225                 230                 235                 240 


Ser Ala Leu Leu Gly Ile Arg Ser Trp Gln Thr Arg Ala Leu Leu Ile 
                245                 250                 255     


Lys Ala Leu Ile Asp Gln Gly Tyr Met Lys Gln Ile Leu Val Ser Asn 
            260                 265                 270         


Asp Trp Leu Phe Gly Phe Ser Ser Tyr Val Thr Asn Ile Met Asp Val 
        275                 280                 285             


Met Asp Arg Val Asn Pro Asp Gly Met Ala Phe Ile Pro Leu Arg Val 
    290                 295                 300                 


Ile Pro Phe Leu Arg Glu Lys Gly Val Pro Gln Glu Thr Leu Ala Gly 
305                 310                 315                 320 


Ile Thr Val Thr Asn Pro Ala Arg Phe Leu Ser Pro Thr Leu Arg Ala 
                325                 330                 335     


Ser 
    


<210>  2
<211>  337
<212>  PRT
<213>  Brevundimonas diminuta

<400>  2

Met Ser Ile Gly Thr Gly Asp Arg Ile Asn Thr Val Arg Gly Pro Ile 
1               5                   10                  15      


Thr Ile Ser Glu Ala Gly Phe Thr Leu Thr His Glu His Ile Cys Gly 
            20                  25                  30          


Ser Ser Ala Gly Phe Leu Arg Ala Trp Pro Glu Phe Phe Gly Ser Arg 
        35                  40                  45              


Lys Ala Leu Ala Glu Lys Ala Val Arg Gly Leu Arg Arg Ala Arg Ala 
    50                  55                  60                  


Ala Gly Val Arg Thr Ile Val Asp Val Ser Thr Phe Asp Ile Gly Arg 
65                  70                  75                  80  


Asp Val Ser Leu Leu Ala Glu Val Ser Arg Ala Ala Asp Val His Ile 
                85                  90                  95      


Val Ala Ala Thr Gly Leu Trp Val Asp Pro Pro Leu Ser Met Arg Leu 
            100                 105                 110         


Arg Ser Val Glu Glu Leu Thr Gln Phe Phe Leu Arg Glu Ile Gln Tyr 
        115                 120                 125             


Gly Ile Glu Asp Thr Gly Ile Arg Ala Gly Ile Ile Lys Val Ala Thr 
    130                 135                 140                 


Thr Gly Lys Ala Thr Pro Phe Gln Glu Leu Val Leu Lys Ala Ala Ala 
145                 150                 155                 160 


Arg Ala Ser Leu Ala Thr Gly Val Pro Val Thr Thr His Thr Ala Ala 
                165                 170                 175     


Ser Gln Arg Asp Gly Glu Gln Gln Ala Ala Ile Phe Glu Ser Glu Gly 
            180                 185                 190         


Leu Ser Pro Ser Arg Val Cys Ile Gly His Ser Asp Asp Thr Asp Asp 
        195                 200                 205             


Leu Ser Tyr Leu Thr Ala Leu Ala Ala Arg Gly Tyr Leu Ile Gly Leu 
    210                 215                 220                 


Asp Gln Ile Pro Phe Ser Ala Ile Gly Leu Glu Asp Asn Ala Ser Ala 
225                 230                 235                 240 


Ser Ala Leu Leu Gly Ile Arg Ser Trp Gln Thr Arg Ala Leu Leu Ile 
                245                 250                 255     


Lys Ala Leu Ile Asp Gln Gly Tyr Met Lys Gln Ile Leu Val Ser Asn 
            260                 265                 270         


Asp Trp Leu Phe Gly Phe Ser Leu Tyr Val Thr Asn Ile Met Asp Val 
        275                 280                 285             


Met Asp Arg Val Asn Pro Asp Gly Met Ala Phe Ile Pro Leu Arg Val 
    290                 295                 300                 


Ile Pro Phe Leu Arg Glu Lys Gly Val Pro Gln Glu Thr Leu Ala Gly 
305                 310                 315                 320 


Ile Thr Val Thr Asn Pro Ala Arg Phe Leu Ser Pro Thr Leu Arg Ala 
                325                 330                 335     


Ser 
    


<210>  3
<211>  337
<212>  PRT
<213>  Brevundimonas diminuta

<400>  3

Met Ser Ile Gly Thr Gly Asp Arg Ile Asn Thr Val Arg Gly Pro Ile 
1               5                   10                  15      


Thr Ile Ser Glu Ala Gly Phe Thr Leu Thr His Glu His Ile Cys Gly 
            20                  25                  30          


Ser Ser Ala Gly Phe Leu Arg Ala Trp Pro Glu Phe Phe Gly Ser Arg 
        35                  40                  45              


Lys Ala Leu Ala Glu Lys Ala Val Arg Gly Leu Arg Arg Ala Arg Ala 
    50                  55                  60                  


Ala Gly Val Arg Thr Ile Val Asp Val Ser Thr Phe Asp Cys Gly Arg 
65                  70                  75                  80  


Asp Val Ser Leu Leu Ala Glu Val Ser Arg Ala Ala Asp Val His Ile 
                85                  90                  95      


Val Ala Ala Thr Gly Leu Trp Val Asp Pro Pro Leu Ser Met Arg Leu 
            100                 105                 110         


Arg Ser Val Glu Glu Leu Thr Gln Phe Phe Leu Arg Glu Ile Gln Tyr 
        115                 120                 125             


Gly Ile Glu Asp Thr Gly Ile Arg Ala Gly Ile Ile Lys Val Ala Thr 
    130                 135                 140                 


Thr Gly Lys Ala Thr Pro Phe Gln Glu Leu Val Leu Lys Ala Ala Ala 
145                 150                 155                 160 


Arg Ala Ser Leu Ala Thr Gly Val Pro Val Thr Thr His Thr Ala Ala 
                165                 170                 175     


Ser Gln Arg Asp Gly Glu Gln Gln Ala Ala Ile Phe Glu Ser Glu Gly 
            180                 185                 190         


Leu Ser Pro Ser Arg Val Cys Ile Gly His Ser Asp Asp Thr Asp Asp 
        195                 200                 205             


Leu Ser Tyr Leu Thr Ala Leu Ala Ala Arg Gly Tyr Leu Ile Gly Leu 
    210                 215                 220                 


Asp Gln Ile Pro Phe Ser Ala Ile Gly Leu Glu Asp Asn Ala Ser Ala 
225                 230                 235                 240 


Ser Ala Leu Leu Gly Ile Arg Ser Trp Gln Thr Arg Ala Leu Leu Ile 
                245                 250                 255     


Lys Ala Leu Ile Asp Gln Gly Tyr Met Lys Gln Ile Leu Val Ser Asn 
            260                 265                 270         


Asp Trp Leu Phe Gly Phe Ser Leu Tyr Val Thr Asn Ile Met Asp Val 
        275                 280                 285             


Met Asp Arg Val Asn Pro Asp Gly Met Ala Phe Ile Pro Leu Arg Val 
    290                 295                 300                 


Ile Pro Phe Leu Arg Glu Lys Gly Val Pro Gln Glu Thr Leu Ala Gly 
305                 310                 315                 320 


Ile Thr Val Thr Asn Pro Ala Arg Phe Leu Ser Pro Thr Leu Arg Ala 
                325                 330                 335     


Ser 
    


<210>  4
<211>  337
<212>  PRT
<213>  Brevundimonas diminuta

<400>  4

Met Ser Ile Gly Thr Gly Asp Arg Ile Asn Thr Val Arg Gly Pro Ile 
1               5                   10                  15      


Thr Ile Ser Glu Ala Gly Phe Thr Leu Thr His Glu His Ile Cys Gly 
            20                  25                  30          


Ser Ser Ala Gly Phe Leu Arg Ala Trp Pro Glu Phe Phe Gly Ser Arg 
        35                  40                  45              


Lys Ala Leu Val Glu Lys Ala Val Arg Gly Leu Arg Arg Ala Arg Ala 
    50                  55                  60                  


Ala Gly Val Arg Thr Ile Val Asp Val Ser Thr Phe Asp Ile Gly Arg 
65                  70                  75                  80  


Asp Val Ser Leu Leu Ala Glu Val Ser Arg Ala Ala Asp Val His Ile 
                85                  90                  95      


Val Ala Ala Thr Gly Leu Trp Val Asp Pro Pro Leu Ser Met Arg Leu 
            100                 105                 110         


Arg Ser Val Glu Glu Leu Thr Gln Phe Phe Leu Arg Glu Ile Gln Tyr 
        115                 120                 125             


Gly Ile Glu Asp Thr Gly Ile Arg Ala Gly Ile Ile Lys Val Ala Thr 
    130                 135                 140                 


Thr Gly Lys Ala Thr Pro Phe Gln Glu Leu Val Leu Arg Ala Ala Ala 
145                 150                 155                 160 


Arg Ala Ser Leu Ala Thr Gly Val Pro Val Thr Thr His Thr Ala Ala 
                165                 170                 175     


Ser Gln Arg Asp Gly Glu Gln Gln Ala Ala Ile Phe Glu Ser Glu Gly 
            180                 185                 190         


Leu Ser Pro Ser Arg Val Cys Ile Gly His Ser Asp Asp Thr Asp Asp 
        195                 200                 205             


Leu Ser Tyr Leu Thr Ala Leu Ala Ala Arg Gly Tyr Leu Ile Gly Leu 
    210                 215                 220                 


Asp Gln Ile Pro Phe Ser Ala Ile Gly Leu Glu Asp Asn Ala Ser Ala 
225                 230                 235                 240 


Ser Ala Leu Leu Gly Asn Arg Ser Trp Gln Thr Arg Ala Leu Leu Ile 
                245                 250                 255     


Lys Ala Leu Ile Asp Gln Gly Tyr Met Lys Gln Ile Leu Val Ser Asn 
            260                 265                 270         


Asp Trp Leu Phe Gly Phe Ser Leu Tyr Val Thr Asn Ile Met Asp Val 
        275                 280                 285             


Met Asp Arg Val Asn Pro Asp Gly Met Ala Phe Ile Pro Leu Arg Val 
    290                 295                 300                 


Ile Pro Phe Leu Arg Glu Lys Gly Val Pro Gln Glu Thr Leu Ala Gly 
305                 310                 315                 320 


Ile Thr Val Thr Asn Pro Ala Arg Phe Leu Ser Pro Thr Leu Arg Ala 
                325                 330                 335     


Ser 
    


<210>  5
<211>  337
<212>  PRT
<213>  Brevundimonas diminuta

<300>
<301>  Ping-Chuan Tsai, Nicholas Fox, Andrew N. Bigley, Steven P. 
       Harvey, David P. Barondeau, and Frank M. Raushel
<302>  Enzymes for the Homeland Defense: Optimizing Phosphotriesterase 
       for the Hydrolysis of Organophosphate Nerve Agents
<303>  Biochemistry
<304>  51
<306>  6463-6475
<307>  2012-07-18

<300>
<301>  Andrew N. Bigley, Chengfu Xu, Terry J. Henderson, Steven P. 
       Harvey, and Frank M. Raushel
<302>  Enzymatic Neutralization of the Chemical Warfare Agent VX: 
       Evolution of Phosphotriesterase for Phosphorothiolate Hydrolysis
<303>  Journal of the American Chemical Society
<304>  135
<306>  10426-10432
<307>  2013-06-21

<400>  5

Met Ser Ile Gly Thr Gly Asp Arg Ile Asn Thr Val Arg Gly Pro Ile 
1               5                   10                  15      


Thr Ile Ser Glu Ala Gly Phe Thr Leu Thr His Glu His Ile Cys Gly 
            20                  25                  30          


Ser Ser Ala Gly Phe Leu Arg Ala Trp Pro Glu Phe Phe Gly Ser Arg 
        35                  40                  45              


Lys Ala Leu Ala Glu Lys Ala Val Arg Gly Leu Arg Arg Ala Arg Ala 
    50                  55                  60                  


Ala Gly Val Arg Thr Ile Val Asp Val Ser Thr Phe Asp Cys Gly Arg 
65                  70                  75                  80  


Asp Val Ser Leu Leu Ala Glu Val Ser Arg Ala Ala Asp Val His Ile 
                85                  90                  95      


Val Ala Ala Thr Gly Leu Trp Val Asp Pro Pro Leu Ser Met Arg Leu 
            100                 105                 110         


Arg Ser Val Glu Glu Leu Thr Gln Phe Phe Leu Arg Glu Ile Gln Tyr 
        115                 120                 125             


Gly Ile Glu Asp Thr Gly Ile Arg Ala Gly Ile Ile Lys Val Ala Thr 
    130                 135                 140                 


Thr Gly Lys Ala Thr Pro Phe Gln Glu Leu Val Leu Lys Ala Ala Ala 
145                 150                 155                 160 


Arg Ala Ser Leu Ala Thr Gly Val Pro Val Thr Thr His Thr Ala Ala 
                165                 170                 175     


Ser Gln Arg Asp Gly Glu Gln Gln Ala Ala Ile Phe Glu Ser Glu Gly 
            180                 185                 190         


Leu Ser Pro Ser Arg Val Cys Ile Gly His Ser Asp Asp Thr Asp Asp 
        195                 200                 205             


Leu Ser Tyr Leu Thr Ala Leu Ala Ala Arg Gly Tyr Leu Ile Gly Leu 
    210                 215                 220                 


Asp Gln Ile Pro Tyr Ser Ala Ile Gly Leu Glu Asp Asn Ala Ser Ala 
225                 230                 235                 240 


Ser Val Leu Met Gly Asn Arg Ser Trp Gln Thr Arg Ala Leu Leu Ile 
                245                 250                 255     


Lys Ala Leu Ile Asp Gln Gly Tyr Met Lys Gln Ile Leu Val Ser Asn 
            260                 265                 270         


Asp Trp Leu Phe Gly Phe Ser Leu Tyr Val Thr Asn Ile Met Asp Val 
        275                 280                 285             


Met Asp Arg Val Asn Pro Asp Gly Met Ala Phe Ile Pro Leu Arg Val 
    290                 295                 300                 


Ile Pro Phe Leu Arg Glu Lys Gly Val Pro Gln Glu Thr Leu Ala Gly 
305                 310                 315                 320 


Ile Thr Val Thr Asn Pro Ala Arg Phe Leu Ser Pro Thr Leu Arg Ala 
                325                 330                 335     


Ser 
    


<210>  6
<211>  337
<212>  PRT
<213>  Brevundimonas diminuta

<400>  6

Met Ser Ile Gly Thr Gly Asp Arg Ile Asn Thr Val Arg Gly Pro Ile 
1               5                   10                  15      


Thr Ile Ser Glu Ala Gly Phe Thr Leu Thr His Glu His Ile Cys Gly 
            20                  25                  30          


Ser Ser Ala Gly Phe Leu Arg Ala Trp Pro Glu Phe Phe Gly Ser Arg 
        35                  40                  45              


Lys Ala Leu Ala Glu Lys Ala Val Arg Gly Leu Arg Arg Ala Arg Ala 
    50                  55                  60                  


Ala Gly Val Arg Thr Ile Val Asp Val Ser Thr Phe Asp Gly Gly Arg 
65                  70                  75                  80  


Asp Val Ser Leu Leu Ala Glu Val Ser Arg Ala Ala Asp Val His Ile 
                85                  90                  95      


Val Ala Ala Thr Gly Leu Trp Val Asp Pro Pro Leu Ser Met Arg Leu 
            100                 105                 110         


Arg Ser Val Glu Glu Leu Thr Gln Phe Phe Leu Arg Glu Ile Gln Tyr 
        115                 120                 125             


Gly Ile Glu Asp Thr Gly Ile Arg Ala Gly Ile Ile Lys Val Ala Thr 
    130                 135                 140                 


Thr Gly Lys Ala Thr Pro Phe Gln Glu Leu Val Leu Lys Ala Ala Ala 
145                 150                 155                 160 


Arg Ala Ser Leu Ala Thr Gly Val Pro Val Thr Thr His Thr Ala Ala 
                165                 170                 175     


Ser Gln Arg Asp Gly Glu Gln Gln Ala Ala Ile Phe Glu Ser Glu Gly 
            180                 185                 190         


Leu Ser Pro Ser Arg Val Cys Ile Gly His Ser Asp Asp Thr Asp Asp 
        195                 200                 205             


Leu Ser Tyr Leu Thr Ala Leu Ala Ala Arg Gly Tyr Leu Ile Gly Leu 
    210                 215                 220                 


Asp Gln Ile Pro Tyr Ser Ala Ile Gly Leu Glu Asp Asn Ala Ser Ala 
225                 230                 235                 240 


Ser Val Leu Met Gly Asn Arg Ser Trp Gln Thr Arg Ala Leu Leu Ile 
                245                 250                 255     


Lys Ala Leu Ile Asp Gln Gly Tyr Met Lys Gln Ile Leu Val Ser Asn 
            260                 265                 270         


Asp Trp Leu Phe Gly Phe Ser Leu Tyr Val Thr Asn Ile Met Asp Val 
        275                 280                 285             


Met Asp Arg Val Asn Pro Asp Gly Met Ala Phe Ile Pro Leu Arg Val 
    290                 295                 300                 


Ile Pro Phe Leu Arg Glu Lys Gly Val Pro Gln Glu Thr Leu Ala Gly 
305                 310                 315                 320 


Ile Thr Val Thr Asn Pro Ala Arg Phe Leu Ser Pro Thr Leu Arg Ala 
                325                 330                 335     


Ser 
    


