Cloning, expression and improvement of catalytic activity of alginate lyase by site-directed mutation
نویسندگان
چکیده
Alginate lyase mainly produces active alginate oligosaccharides (AOS) by degrading via β-elimination process. In this study, the Pseudoalteromonas sp. Alg6B lyase-encoding gene alg6B-7 from polysaccharide (PL)-7 family was successfully cloned, sequenced, expressed in Escherichia coli. Based on rational design and amino acid sequence alignment of various sources, four positive mutants were obtained. The specific enzyme activities I62A, A99K, V132S, L157T 38.84%, 42.85%, 75.8% 51.83% higher than that wild enzyme, respectively. Kcat/Km values both increased, catalytic efficiency V132S 1.92-fold especially. employed study achieved dramatic improvement activity, which may provide application potential industrial production.
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ژورنال
عنوان ژورنال: Systems microbiology and biomanufacturing
سال: 2022
ISSN: ['2662-7663', '2662-7655']
DOI: https://doi.org/10.1007/s43393-022-00084-w