Fang_2024_J.Biosci.Bioeng__

Reference

Title : Characterization of a novel carboxylesterase from Streptomyces lividans TK24 and site-directed mutagenesis for its thermostability - Fang_2024_J.Biosci.Bioeng__
Author(s) : Fang J , An L , Yu J , Ma J , Zhou R , Wang B
Ref : J Biosci Bioeng , : , 2024
Abstract :

As an industrial enzyme that catalyzes the formation and cleavage of ester bonds, carboxylesterase has attracted attention in fine chemistry, pharmaceutical, biological energy and bioremediation fields. However, the weak thermostability limits their further developments in industrial applications. In this work, a novel carboxylesterase (EstF) from Streptomyces lividans TK24, belonging to family XVII, was acquired by successfully heterologous expressed and biochemically identified. The EstF exhibited optimal activity at 55 degreesC, pH 9.0 and excellent catalytic performances (K(m) = 0.263 mM, k(cat)/K(m) = 562.3 s(-1) mM(-1) for p-nitrophenyl acetate (pNPA(2)) hydrolysis). Besides, the EstF presented exceptionally high thermostability with a half-life of 387.23 h at 55 degreesC and 2.86 h at 100 degreesC. Furthermore, the EstF was modified to obtain EstF(P144G) using the site-directed mutation technique to investigate the effect of single glycine on thermostability. Remarkably, the mutant EstF(P144G) displayed a 5.10-fold increase of half-life at 100 degreesC versus wild-type without affecting catalytic performance. Structural analysis implied that the glycine introduction could release a steric strain and induce cooperative effects between distal residues to increase the thermostability. Therefore, the thermostable EstF and EstF(P144G) with prominently catalytic characteristics have potential industrial applications and the introduction of a single glycine strategy opens up alternative avenues for the thermostability engineering of other enzymes.

PubMedSearch : Fang_2024_J.Biosci.Bioeng__
PubMedID: 38871580
Gene_locus related to this paper: strco-SCO7396

Related information

Gene_locus strco-SCO7396

Citations formats

Fang J, An L, Yu J, Ma J, Zhou R, Wang B (2024)
Characterization of a novel carboxylesterase from Streptomyces lividans TK24 and site-directed mutagenesis for its thermostability
J Biosci Bioeng :

Fang J, An L, Yu J, Ma J, Zhou R, Wang B (2024)
J Biosci Bioeng :