Chiang Mai Journal of Science

Print ISSN: 0125-2526 | eISSN : 2465-3845

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Heterologous Expression, and Characterization of a Thermophilic and Acid-Tolerant GH10 Xylanase from Thermomonospora amylolytica

Lei Li †, Lin-Hua Li†, Wei Hu, Yang-Min Li, Mao-Song Li, Zheng-Feng Yang, Dorji Phurbu, Yan-Yan Zheng, Syed Raziuddin Quadri, Manik Prabhu Narsing Rao* and Yi-Rui Yin*
* Author for corresponding; e-mail address: yinyirui@dali.edu.cn, deene.manik@gmail.com
ORCID ID: https://orcid.org/0000-0002-4777-3452
Volume: Vol.53 No.6 (November 2026: In progress)
Research Article
DOI: https://doi.org/10.12982/CMJS.2026.106
Received: 5 April 2026, Revised: 5 September 2026, Accepted: 19 September 2026, Published: -

Citation: Li L., Li L.-H., Hu W., Li Y.-M., Li M.-S., Yang Z.-F., et al., Heterologous expression, and characterization of a thermophilic and acid-tolerant GH10 xylanase from Thermomonospora amylolytica. Chiang Mai Journal of Science, 2026; 53(6): e2026106. DOI 10.12982/CMJS.2026.106.

Graphical Abstract

Graphical Abstract

Abstract

Xylanases, a subclass of glycoside hydrolases, catalyze the breakdown of xylan into xylan disaccharides, oligosaccharides, and xylose, playing a critical role in the degradation and utilization of hemicelluloses. However, extreme conditions hinder their broader industrial applications. In this study, a GH10 xylanase gene (referred to as taxyn10a) was sourced from Thermomonsospora amylolytica, cloned, and heterologously expressed in Escherichia coli BL21(DE3). The recombinant TaXyn10A was purified using Ni2+-affinity chromatography. Its optimal temperature and pH were determined to be 65°C and 5.6, respectively. The enzyme maintained over 80% of its activity between 55-70°C, with about 60% residual activity after 120 minutes at 60°C. Additionally, it preserved more than 80% of its activity in a pH range of 3.6-6.6. TaXyn10A exhibited over 60% activity in the presence of 1 or 10 mM ions (K+, Fe3+, Mg2+, Ca2+, Zn2+, Mn2+, Ni2+, Co2+, Pb2+, Cu2+), as well as 1% EDTA, PMSF, and DTT. Kinetic analysis with beechwood xylan yielded apparent Vmax, Km, and kcat values of 69.44 μmol min⁻¹ mg⁻¹, 24.53 mg mL⁻¹, and 53.35 s⁻¹, respectively. These findings demonstrate that TaXyn10A was active over moderately acidic pH and elevated-temperature ranges and tolerates various metal ions and chemical additives, highlighting its potential for feed processing and lignocellulosic biomass conversion.

Keywords: Thermomonospora amylolytica, thermophilic, thermotolerant, acid-tolerant, xylanase, lactic acid, Rhizopus oryzae, pretreatment, optimization, response surface methodology (RSM), central composite design (CCD), agro-industrial waste

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