Directed evolution of microbial oxidative enzymes

Current Opinion in Biotechnology

Volumn 11, Issue 3, 2000, Pages 250-254

  Cherry, Joel R ^a  

^a NOVO NORDISK INC   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

BACTERIAL ENZYME; CATALASE; CYTOCHROME P450; PEROXIDASE;

ENZYME MECHANISM; ENZYME SPECIFICITY; EVOLUTION; NONHUMAN; OXIDATION; PH; PRIORITY JOURNAL; SHORT SURVEY; THERMOSTABILITY;

BACTERIA (MICROORGANISMS);

EID: 0034079648     PISSN: 09581669     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0958-1669(00)00092-6     Document Type: Review

References (19)

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8. ••] describes the mutagenesis of a catalase gene to shift its reaction specificity to that of a peroxidase in two successive rounds. This is one of very few examples of the conversion of an enzymatic reaction pathway by mutagenesis.
9. 2). It is notable in that it addresses an industrial problem (laundry dye transfer) using a combination of rational, random, and shuffling techniques. It is also the first described use of the yeast in vivo recombination system for the shuffling of synthetic variants.
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14. •,13], this manuscript is important in that it describes a novel screening strategy that avoids the problem of cofactor recycling. Although the improvements in P450 activity were modest, this is a landmark work in its approach to oxidase evolution.
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16. Kumamaru T., Suenaga H., Mitsuoka M., Watanabe T., Furukawa K. Enhanced degradation of polychlorinated biphenyls by directed evolution of biphenyl dioxygenase. Nat Biotechnol. 16:1998;663-666. A very nice demonstration of the power of shuffling in which two genes with differing substrate specificities were recombined to generate dioxygenases with novel properties.
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19. Kikuchi M., Ohnishi K., Harayama S. Novel family shuffling methods for the in vitro evolution of enzymes. Gene. 236:1999;159-167. Family shuffling of two dioxygenases yielded mutants with 13- and 26-fold higher thermostability than the parents. The authors demonstrate that, in this case, the efficiency of PCR reassembly of fragmented genes is better when the genes are fragmented using restriction enzymes rather than DNase I.