Characterization of glycoside hydrolase family 11 xylanase from Streptomyces sp. strain J103; its synergetic effect with acetyl xylan esterase and enhancement of enzymatic hydrolysis of lignocellulosic biomass

• Published in: Microbial cell factories 2021-07, Vol.20 (1), p.1-129, Article 129
• Main Authors: Marasinghe, Svini Dileepa, Jo, Eunyoung, Hettiarachchi, Sachithra Amarin, Lee, Youngdeuk, Eom, Tae-Yang, Gang, Yehui, Kang, Yoon-Hyeok, Oh, Chulhong
• Format: Article
• Language: English
• Subjects: Amino acids; Antibiotics; Bacteria; Biodegradation; Biodiesel fuels; Biofuels; Biomass; Carbon dioxide; Cellulase; Cellulose; Chromatography; Cobalt; Column chromatography; Degradation; E coli; Enzymes; Esterase; Esterification; Experiments; Expression; Genetic aspects; Glycosidases; Glycoside hydrolase; Glycosides; Hydrolase; Hydrolases; Hydrolysis; Industrial applications; Industrial microorganisms; Lignocellulose; Lignocellulosic biomass; Manganese ions; Methods; Microbial enzymes; Molecular weight; Peptides; Physiological aspects; Production processes; Protein biosynthesis; Protein synthesis; Proteins; Purification; Streptomyces; Streptomyces sp. strain J103; Synergism; Thermal stability; Xylan; Xylanase
• ISSN: 1475-2859; 1475-2859
• DOI: 10.1186/s12934-021-01619-x

Xylanase-containing enzyme cocktails are used on an industrial scale to convert xylan into value-added products, as they hydrolyse the [beta]-1,4-glycosidic linkages between xylopyranosyl residues. In the present study, we focused on xynS1, the glycoside hydrolase (GH) 11 xylanase gene derived from the Streptomyces sp. strain J103, which can mediate XynS1 protein synthesis and lignocellulosic material hydrolysis. xynS1 has an open reading frame with 693 base pairs that encodes a protein with 230 amino acids. The predicted molecular weight and isoelectric point of the protein were 24.47 kDa and 7.92, respectively. The gene was cloned into the pET-11a expression vector and expressed in Escherichia coli BL21(DE3). Recombinant XynS1 (rXynS1) was purified via His-tag affinity column chromatography. rXynS1 exhibited optimal activity at a pH of 5.0 and temperature of 55 [degrees]C. Thermal stability was in the temperature range of 50-55 [degrees]C. The estimated K.sub.m and V.sub.max values were 51.4 mg/mL and 898.2 U/mg, respectively. One millimolar of Mn.sup.2+ and Na.sup.+ ions stimulated the activity of rXynS1 by up to 209% and 122.4%, respectively, and 1 mM Co.sup.2+ and Ni.sup.2+ acted as inhibitors of the enzyme. The mixture of rXynS1, originates from Streptomyces sp. strain J103 and acetyl xylan esterase (AXE), originating from the marine bacterium Ochrovirga pacifica, enhanced the xylan degradation by 2.27-fold, compared to the activity of rXynS1 alone when Mn.sup.2+ was used in the reaction mixture; this reflected the ability of both enzymes to hydrolyse the xylan structure. The use of an enzyme cocktail of rXynS1, AXE, and commercial cellulase (Celluclast[R] 1.5 L) for the hydrolysis of lignocellulosic biomass was more effective than that of commercial cellulase alone, thereby increasing the relative activity 2.3 fold. The supplementation of rXynS1 with AXE enhanced the xylan degradation process via the de-esterification of acetyl groups in the xylan structure. Synergetic action of rXynS1 with commercial cellulase improved the hydrolysis of pre-treated lignocellulosic biomass; thus, rXynS1 could potentially be used in several industrial applications.