Dated: Jun 01, 2022
Publication Name: Food Bioscience
Citations : Engineering D-allulose 3-epimerase from Clostridium cellulolyticum for improved thermostability using directed evolution facilitated by a nonenzymatic colorimetric screening assay
D-Allulose 3-epimerase (DAEase) is a biocatalyst of concern for D-allulose enzymatic synthesis; however, it exhibits comparatively low thermal stability. To facilitate the directed evolution of DAEase, a nonenzymatic colorimetric assay was developed for high-throughput screening of enhanced DAEase mutants. One desirable mutant, MT4 (S38F/F42N/A70P/T119P), was screened based on purifying selection of the randomly mutagenesis library and positive screening of the randomly shuffled library. Compared to the wild-type, the combinational mutant MT4 had higher catalytic activity (1.23-fold) and showed 5, 6.2, and 6.92 °C increases in Topt, T50, and Tm values, respectively.
Contributors: Jiajun Chen a b, Zhaolin Huang a b, Ting Shi c, Dawei Ni a b, Yingying Zhu a b, Wei Xu a b, Wenli Zhang a b, Wanmeng Mu