Enhancing thermostability of xylanase from rumen microbiota by molecular cyclization.

Kexin Zhou; Huan Wang; Xintao Zhu; Anqi Zheng; Nuo LI; Xiaobao Sun; Deying Gao; Peipei AN; Jiakun Wang; Guoying Qian; Qian Wang

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Enhancing thermostability of xylanase from rumen microbiota by molecular cyclization.

Author: Kexin ZHOU ¹ ; Huan WANG ¹ ; Xintao ZHU ¹ ; Anqi ZHENG ¹ ; Nuo LI ¹ ; Xiaobao SUN ¹ ; Deying GAO ¹ ; Peipei AN ² ; Jiakun WANG ² ; Guoying QIAN ¹ ; Qian WANG ¹
Author Information

1. College of Biological & Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, Zhejiang, China.
2. College of Animal Sciences, Zhejiang University, Hangzhou 310012, Zhejiang, China.
Publication Type:Journal Article
Keywords: molecular cyclization; rumen; substrate degradation; thermostability; xylanase
MeSH: Animals; Cyclization; Endo-1,4-beta Xylanases; chemistry; metabolism; Enzyme Stability; Industrial Microbiology; methods; Microbiota; Protein Engineering; Rumen; enzymology; microbiology; Temperature
From: Chinese Journal of Biotechnology 2020;36(5):920-931
CountryChina
Language:Chinese
Abstract: The capacity for thermal tolerance is critical for industrial enzyme. In the past decade, great efforts have been made to endow wild-type enzymes with higher catalytic activity or thermostability using gene engineering and protein engineering strategies. In this study, a recently developed SpyTag/SpyCatcher system, mediated by isopeptide bond-ligation, was used to modify a rumen microbiota-derived xylanase XYN11-6 as cyclized and stable enzyme C-XYN11-6. After incubation at 60, 70 or 80 ℃ for 10 min, the residual activities of C-XYN11-6 were 81.53%, 73.98% or 64.41%, which were 1.48, 2.92 or 3.98-fold of linear enzyme L-XYN11-6, respectively. After exposure to 60-90°C for 10 min, the C-XYN11-6 remained as soluble in suspension, while L-XYN11-6 showed severely aggregation. Intrinsic and 8-anilino-1-naphthalenesulfonic acid (ANS)-binding fluorescence analysis revealed that C-XYN11-6 was more capable of maintaining its conformation during heat challenge, compared with L-XYN11-6. Interestingly, molecular cyclization also conferred C-XYN11-6 with improved resilience to 0.1-50 mmol/L Ca²⁺ or 0.1 mmol/L Cu²⁺ treatment. In summary, we generated a thermal- and ion-stable cyclized enzyme using SpyTag/SpyCatcher system, which will be of particular interest in engineering of enzymes for industrial application.