Enhancing Catalytic Efficiency of an Actinoplanes utahensis Echinocandin B Deacylase through Random Mutagenesis and Site-Directed Mutagenesis View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2019-11-18

AUTHORS

Ying-Nan Cheng, Shuai Qiu, Feng Cheng, Chun-Yue Weng, Ya-Jun Wang, Yu-Guo Zheng

ABSTRACT

Echinocandin B deacylase (EBDA), from Actinoplanes utahensis ZJB-08196, is capable of cleaving the linoleoyl group from echinocandin B (ECB), forming the echinocandin B nucleus (ECBN), which is a key precursor of semisynthetic antifungal antibiotics. In the present study, molecular evolution of AuEBDA by random mutagenesis combined with site-directed mutagenesis (SDM) and screening was performed. Random mutagenesis on the wild-type (WT) AuEBDA generated two beneficial substitutions of G287Q, R527V. The “best” variant AuEBDA-G287Q/R527V was obtained by combining G287Q with R527V through SDM, which was most active at 35 °C, pH 7.5, with Km and vmax values of 0.68 mM and 395.26 U/mg, respectively. Mutation of G287Q/R527V markedly increased the catalytic efficiency kcat/Km by 290% compared with the WT-AuEBDA. More... »

PAGES

1257-1270

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s12010-019-03170-3

DOI

http://dx.doi.org/10.1007/s12010-019-03170-3

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1122678947

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/31741208


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