Enhancement of thermostability of kojibiose phosphorylase from Thermoanaerobacter brockii ATCC35047 by random mutagenesis
作者： Takuo Yamamoto , Kazuhisa Mukai , Hiroshi Yamashita , Michio Kubota , Shigeharu Fukuda
DOI: 10.1263/JBB.100.212
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摘要: Random mutation by error-prone PCR was introduced into kojibiose phosphorylase from Thermoanaerobacter brockii ATCC35047. One thermostable mutant enzyme, D513N, isolated. The D513N enzyme showed an optimum temperature of 67.5–70°C (the wild type, 65°C), and thermostability up to 67.5°C 60°C). half-lives were estimated be 135 h at 60°C, 110 min 70°C 6 75°C, respectively. They about 1.6-fold, 7-fold 6-fold longer than those the wild-type
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