The Î±-L-rhamnosidase BtRha from Bacteroides thetaiotao VPI-5482 is a specific enzyme that selectively hydrolyzes the Î±-1,2 glycosidic bond between rhamnose and rhamnose, allowing the bioconversion of epimedin C to icariin. In this study, BtRha was molecularly modified using B-factor-saturation mutagenesis strategy and the introduction of disulfide bonds, resulting in a mutant with significantly improved catalytic efficiency, S592C, and two thermally stable mutants, E39W and E39W-S592C. The results showed that the half-lives of E39W and E39W-S592C at 55 Â°C were 10.4 and 9.4-fold higher, respectively, than that of the original enzyme, The mutant S592C showed a 63.3% reduction in Km value and a 163.6% increase in catalytic efficiency (kcat/Km value), which improved the ability to hydrolyze epimedin C to icariin effectively. In addition, high-level expression of Î±-L-rhamnosidase mutant S592C was established. With 0.1 mM IPTG as an inducer, induction temperature of 32 Â°C, induction pH...
Modification to increase the thermostability and catalytic efficiency of α-L-rhamnosidase from Bacteroides thetaiotaomicron and high-level expression.
Linguo Zhao, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, 159 Long Pan Road, Nanjing, 210037, China. E-mail firstname.lastname@example.org
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Changning Lu, Yurong Dong, Kaixuan Ke, Kang Zou, Zhenzhong Wang, Wei Xiao, Jianjun Pei, Linguo Zhao; Modification to increase the thermostability and catalytic efficiency of α-L-rhamnosidase from Bacteroides thetaiotaomicron and high-level expression.. IFIS Food and Health Sciences Database 2022; doi:
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