Characterization of a novel highly thermostable esterase from the Gram‐positive soil bacterium Streptomyces lividans TK64

Abstract

A novel esterase gene (estW) from soil bacterium Streptomyces lividans TK64 was successfully cloned using a pair of homologous primers. The estW gene encoded a protein (EstW) of 289 amino acid residues with a predicted molecular weight of 31.43 kDa. Sequence alignment revealed that EstW show relatively high levels of homology to other lipolytic enzymes characterized from Streptomyces and phylogenetic analysis suggested EstW belongs to the bacterial lipase/esterase family I. The estW gene was expressed at a high level in Escherichia coli and the recombinant enzyme was purified to homogeneity. The purified EstW was characterized via hydrolysis of various p-nitrophenyl esters and the best substrate was found to be p-nitrophenyl acetate (pNPA). Maximal activity of the recombinant protein was observed at pH 8.0 and 50 °C with pNPA as the substrate. The calculated activation energy (Ea ) of the esterase reaction was 9.12 kcal/mol. Half-life of EstW at 95 °C was approximately 12.5 H, making it the most thermostable esterase among all of the known lipolytic enzymes from Streptomyces, and the thermostability of EstW was similar to those of some enzymes characterized from the thermophilic bacteria. EstW exhibited relatively high tolerance to several detergents and required no cations for its maximal activity. The unique properties of EstW, namely its high thermostability and stability in the presence of organic solvents, may render it a potential candidate for industrial applications.

In silico Screening and Heterologous Expression of a Polyethylene Terephthalate Hydrolase (PETase)-Like Enzyme (SM14est) With Polycaprolactone (PCL)-Degrading Activity, From the Marine Sponge-Derived Strain Streptomyces sp. SM14

“…Several Streptomyces genes encoding different enzymes have previously been heterologously expressed in E. coli (Spasic et al, 2018) – including a xylanase from S. mexicanus HY-14 JQ943651 (Kim et al, 2014); a laccase from S. coelicolor (Sherif et al, 2013); a protease from S. koyangensis (Ben Elhoul et al, 2015); a glucose isomerase from Streptomyces sp. SK (Ben Hlima et al, 2013); and an esterase from S. lividans (Wang et al, 2016). In these cases however, unlike with the heterologous expression of the SM14est in E. coli , the native signal sequence was not employed in the expression constructs.…”

A Novel Lipase from Streptomyces exfoliatus DSMZ 41693 for Biotechnological Applications

“…Evolutionary analyses were conducted in MEGA11 [65]. [62]; SlEst: esterase from S. lividans (ACX47356.1) [63]; SsEst: esterase from S. scabies [64]. The evolutionary history was inferred using the neighbor-joining method.…”

Revisiting the scope and applications of food enzymes from extremophiles

“…For instance, Candida antarctica lipase B (Cal B) acts as a lipase when adsorbed to an acylglyceride interface and as TA B L E 3 Some esterase producing extremophiles an esterase when exposed to an aqueous environment (Benson & Pleiss, 2017). Several microbial esterases from extreme environments (Table 3) are very active at low and high temperature, salt concentration, and pH (Adıgüzel, 2020;Curci et al, 2019;Jiang, Zhang, Gao, & Hu, 2016;Polkade, Ramana, Joshi, Pardesi, & Shouche, 2015;Wang, Wang, Cao, & Zhu, 2016;Zarafeta et al, 2016).…”