A glucotolerant β-glucosidase from the fungus Talaromyces amestolkiae and its conversion into a glycosynthase for glycosylation of phenolic compounds.

Microbial Cell Factories
Juan Antonio Méndez-LíterMaría Jesús Martínez

Abstract

The interest for finding novel β-glucosidases that can improve the yields to produce second-generation (2G) biofuels is still very high. One of the most desired features for these enzymes is glucose tolerance, which enables their optimal activity under high-glucose concentrations. Besides, there is an additional focus of attention on finding novel enzymatic alternatives for glycoside synthesis, for which a mutated version of glycosidases, named glycosynthases, has gained much interest in recent years. In this work, a glucotolerant β-glucosidase (BGL-1) from the ascomycete fungus Talaromyces amestolkiae has been heterologously expressed in Pichia pastoris, purified, and characterized. The enzyme showed good efficiency on p-nitrophenyl glucopyranoside (pNPG) (Km= 3.36 ± 0.7 mM, kcat= 898.31 s-1), but its activity on cellooligosaccharides, the natural substrates of these enzymes, was much lower, which could limit its exploitation in lignocellulose degradation applications. Interestingly, when examining the substrate specificity of BGL-1, it showed to be more active on sophorose, the β-1,2 disaccharide of glucose, than on cellobiose. Besides, the transglycosylation profile of BGL-1 was examined, and, for expanding its synthetic cap...Continue Reading

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Citations

Mar 31, 2021·Biotechnology Advances·Zuzana MészárosKristýna Slámová
Apr 20, 2021·Applied Biochemistry and Biotechnology·Ravish GodseRam Kulkarni
Aug 16, 2021·Carbohydrate Research·Gregory S BulmerJolanda M van Munster
Aug 22, 2021·International Microbiology : the Official Journal of the Spanish Society for Microbiology·Alicia PrietoMaría Jesús Martínez
Sep 1, 2021·Biotechnology Advances·Zuzana MészárosKristýna Slámová

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Methods Mentioned

BETA
glycosylation
Nuclear Magnetic Resonance
NMR
transglycosylation
PCR

Software Mentioned

- Ease
Masshunter Data Acquisition
clustal omega
BLASTN
SignalP
TBLASTN
SigmaPlot
Stat
Masshunter Qualitative Analysis

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