Table 1.
Enzymes and their mutants formed by site directed mutagenesis.
S. no. | Source | Wild type enzyme | Substrate specificity/cofactor specificity | Mutant enzyme | New substrate specificity/cofactor specificity | Mutations at specific sites | References |
---|---|---|---|---|---|---|---|
1. | Trichomonas vaginalis | Lactate dehydrogenase | Lactate etc. | Malate dehydrogenase | Malate etc. | Leu91Arg | Wu et al., 1999 |
2. | Citrobacter freundii | Tyrosine phenol lyase | β- elimination of tyrosine | Dicarboxylic amino acid β-lyase enzyme | Reversible transfer of an amino group from dicarboxylic amino acids to oxo acids | Arg100Thr, Val283Arg | Mouratou et al., 1999 |
3. | Citrobacter freundii | Tyrosine phenol lyase | β- elimination of tyrosine | Tyrosine phenol lyase | Decreased affinity for β- elimination of tyrosine | Asn185Ala, Tyr71Phe, Thr124Asp, Phe448His | Chen H. Y. et al., 1995; Barbolina et al., 2000; Demidkina et al., 2002 |
4. | Lactobacillus pentosus | NAD-dependent D-lactate dehydrogenase | Pyruvate | D-2hydroxyisocaproate dehydrogenase | Larger aliphatic or aromatic 2-ketoacid substrates | Tyr52Leu | Tokuda et al., 2003 |
5. | E. coli | Choline acetyl transferase | Acetyl group acceptor is choline | Carnitine acetyl transferase | Specificity More for carnitine and less for choline | Val459Thr, Asp460Glu, Asn461Thr Asn514Arg | Reznik et al., 1998 |
6. | E. coli | Aspartate amino- transferase | reversible transfer of the amino group of aspartate or glutamate to the cognate oxo acids | L-aspartate-β decarboxylase | β-caboxylase activity | Tyr225Arg, Arg292Lys Arg386Ala | Graber et al., 1999 |
7. | Bacillus stearothermophilus | Lactate dehydrogenase | specific for cofactor NAD+ | Same | Utilized NADP+ far better | Phe16Gln, Cys81Ser Asn85Arg | Flores and Ellington, 2005 |
8. | E. coli | β-Glucuronidase | β- galactosidase activity | Same | Increased in β galactosidase activity | Thr509Ala, Ser557Pro, Asn566Ser Lys568Gln | Geddie and Matsumura, 2004 |
9. | E. coli | Malate dehydrogenase | Inter-conversion of malate & oxaloacetate | Same | Catalytic activity is decreased & specificity for other substrate increased | Arg153Cys | Wright and Viola, 2001 |
10. | E. coli | Aspartate amino transferases | Aspartate | Same | Five-fold increase in activity | Asn34Asp, Ile37Met, Ser139Gly, Asn142Thr, Asn297Ser, Val387Leu | Yano et al., 1998 |
11. | E. coli | Aspartate amino transferases | Aspartate | same | Valine | 17 mutations | Oue et al., 1999 |
12. | E. coli | Streptavidin | Biotin | Same | Biotin analogs e.g., Iminobiotin | Asn23Ala Ser27Asp | Reznik et al., 1998 |
13. | Klebsiella pneumoniae | 1,2 propanediol oxidoreductase | Coenzyme-NADH | Same | Coenzyme-NADH and NADPH | Asp41Ala, Asp41Gly | Ma et al., 2010 |
14. | Bacillus lentus | Subtilisins | Proteolytic activity | Same | Increased proteolytic activity | Lys27Arg/ Asn87Ser/ Val104Tyr/ Asn123Ser /Thr274Ala Asn76Asp/ Asn87Ser/ Ser103Ala/ Val104Ile | Graycar et al., 1999 |
15. | Streptomyces spp. | Cholestrol oxidase | Pregnenolone, cholesterol | Same | Increased catalytic activity | Ser379Thr | Toyama et al., 2002 |
16. | E. coli | Isocitrate dehydrogenase | Coenzyme-NADP | Same | Coenzyme-NAD | 6 mutations | Chen R. et al., 1995 |
17. | Peptostreptococcus asaccharolyticus | Glutamate dehydrogenase | Coenzyme-NADH | Same | Coenzyme-NADPH | Glu243Lys/Glu243Asp | Carrigan and Engel, 2007 |
18. | Candida boidinii | Formate dehydrogenase | Coenzyme-NAD+ | Same | Coenzyme-NADP+ | Asp195Gln/Tyr196His | Andreadeli et al., 2008 |
19. | E. coli | Malate dehydrogenase | Oxaloacetate | Phenylactate dehydrogenase | Phenyl pyruvate | Arg81Cys | Wright et al., 2000 |
20. | Streptomyces coelicolor | Malate dehydrogenase | NADP | same | NADPH | Glu42Gly, Ile43Ser, Pro45Arg, Ala46Ser | Ge et al., 2014 |
21. | Pseudomonas N176 | Glutaryl-7-ACA acylase | CephC | CephalosporinCacylases (CA) | CephC | His57Ser His70Ser, Leu154Tyr | Conti et al., 2014 |
22. | Candida tenuis | Xylose reductase | NADPH | Same | NADH | Lys274Arg–Asn276Asp | Petschacher et al., 2005 |
23. | Thermus thermophilus | Lactate dehydrogenase | NADH | Same | NADPH | 7 amino acid residues | Tomita et al., 2006b |
24. | Pichia stipitis | Xylose reductase | NADPH | Same | NADH | Lys21Ala/ Asn272Asp | Zeng et al., 2009 |
25. | Clostridium symbiosum | Glutamate dehydrogenase | NADH | Same | NADPH | Asp263Lys, Phe238Ser, Pro262Ser | Griffin and Engel, 2011 |
26. | Thermus flavus | Malate dehydrogenase | NADH | Same | NADPH | 7 amino acids residues | Nishiyama et al., 1993; Tomita et al., 2006a |
27. | E. coli | Lactaldehyde dehydrogenase | NADH | Same | NADPH | Phe180Thr | Rodríguez-Zavala, 2008 |
28. | Candida magnoliae | Carbonyl reductase | NADPH | Same | NADH | 8 amino acid residues | Morikawa et al., 2005 |
29. | Neurospora crassa | Nitrate reductase | NADPH | Same | NADH | Ser920Asp Arg932 Asp | Shiraishi et al., 1998 |
30. | Lactobacillus bulgaricus | Lactate dehydrogenase | NADH | Same | NADPH | Asp175Ala | Bernard et al., 1995 |
31. | Glomerella cingulata | Cutinase | Hydrolysis of esters and triglycerides | Same | Catalytically inactive | His204Asn | Nyon et al., 2009 |
32. | Pseudomonas aeruginosa LST- 03 | Lipase | Ester synthesis and inter-esterification reaction and lipid hydrolysis | Same | Increased organic solvent stability | Ser155Leu Gly157Arg, Ser164Lys, Ser194Arg, Asp209Asn | Kawata and Ogino, 2010 |
33. | Geobacillus kaustophilus HTA426 | Lactonase | 3-oxo-N-dodecanoyl-L-homoserine lactone | Same | 72-fold increase in the catalytic efficiency | Glu101Asn/ Arg230Ile | Chow et al., 2010 |
3-oxo-N-dodecanoyl-L-homoserine lactone | Same | N-butyryl-l-homoserine lactone | Asp266Asn | Chow et al., 2010 | |||
34. | alkalophilic Bacillus sp. | Cyclodextrin glucano transferase | produce cyclodextrins from starch | Same | higher product yields | His233Tyr | Leemhuis et al., 2010 |
35. | E. coli | Lactaldehyde reductase | NADH | Same | 3.6-fold increase in kcat | Met185Cys | Cahn et al., 2016 |
36. | Saccharomyces cerevisae | Cinnamyl alcohol dehydrogenase | NAD(P)H | Same | 82-fold increase in activity | Gln110Val | Cahn et al., 2016 |
37. | Actinoplanes utahensis | Cephalosporin acyclase | Cephalosporin | Same | Aculeacin A | Phe177(β)Gly/Met145(α)Ala and Phe177(β)Gly/Met145(α)Ala/Tyr149(α)Val | Isogai and Nakayama, 2016 |