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Enhanced catalytic efficiency of CotA-laccase by DNA shuffling.

ABSTRACT: Bacterial CotA-laccases exhibit higher activity in alkaline pH and salt concentration conditions compared to laccases from white-rot fungi. They are considered as green catalysts in decolorizing of industrial dyes. However, CotA-laccases are limited due to the low yield and catalytic efficiency as the spore-bound nature of CotA. A DNA shuffling strategy was applied to generate a random mutation library. To improve laccase activities, a mutant (T232P/Q367R 5E29) with two amino acid substitutions was identified. The catalytic efficiency of mutant 5E29 was 1.21 fold higher compared with that of the wild-type. The Km and kcat values of 5E29 for SGZ were of 20.3 ± 1.3 µM and 7.6 ± 2.7 s-1. The thermal stability was a slight enhancement. Indigo Carmine and Congo red were efficiently decolorized by using this mutant at pH 9.0. These results provide that 5E29 CotA-laccase is a good candidate for biotechnology applications under alkaline condition, with an effective decolorization capability.

SUBMITTER: Ouyang F

PROVIDER: S-EPMC6550543 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

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Enhanced catalytic efficiency of CotA-laccase by DNA shuffling.

Ouyang Fengju F Zhao Min M

Bioengineered 20191201 1

Bacterial CotA-laccases exhibit higher activity in alkaline pH and salt concentration conditions compared to laccases from white-rot fungi. They are considered as green catalysts in decolorizing of industrial dyes. However, CotA-laccases are limited due to the low yield and catalytic efficiency as the spore-bound nature of CotA. A DNA shuffling strategy was applied to generate a random mutation library. To improve laccase activities, a mutant (T232P/Q367R 5E29) with two amino acid substitutions ...[more]

PMID: 31142180

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