食品与发酵工业 >

2026 , Vol. 52 >Issue 16: 17 - 24

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.046685

精准发酵

嗜热玫瑰红球菌来源葡萄糖/木糖激活型高温β-葡萄糖苷酶的重组表达与酶学性质研究

  • 刘洋 ,
  • 黄洁 ,
  • 张涛 ,
  • 窦康杰 ,
  • 徐有山 ,
  • 赵维萍
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  • 1(滁州学院 生物与食品工程学院,安徽 滁州,239000)
    2(安徽大学 生命科学与医学工程学院,安徽 合肥,230601)
    3(天长市圣丰生态农业科技有限公司,安徽 滁州,239000)
第一作者:博士,副教授(赵维萍实验师为通信作者,E-mail:zwp_1020@163.com)

收稿日期: 2026-03-26

  修回日期: 2026-04-08

  网络出版日期: 2026-09-10

基金资助

安徽省高等学校自然科学研究项目(2023AH051596,KJ2019A0639,KJ2020A0715);安徽省高校理工科教师赴企业挂职实践计划项目(2024jsqygz111);滁州学院科学研究项目(2022XJZD21);企业横向项目(HX2022257,HX2022258)

收起

Recombinant expression and enzymatic properties of a glucose- and xylose-stimulated thermophilic β-glucosidase from Thermomicrobium roseum

  • LIU Yang ,
  • HUANG Jie ,
  • ZHANG Tao ,
  • DOU Kangjie ,
  • XU Youshan ,
  • ZHAO Weiping
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  • 1(School of Biological and Food Engineering, Chuzhou University, Chuzhou 239000, China)
    2(School of Life Sciences and Medical Engineering, Anhui University, Hefei 230601, China)
    3(Tianchang Shengfeng Eco-Agricultural Technology Co.Ltd., Chuzhou 239000, China)

Received date: 2026-03-26

  Revised date: 2026-04-08

  Online published: 2026-09-10

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摘要

针对β-葡萄糖苷酶在应用中普遍存在的催化活性偏低、不耐高温以及酶活力容易受到水解产物的反馈抑制等问题,该研究从嗜热玫瑰红球菌(Thermomicrobium roseum)中筛选获得了一个新型的β-葡萄糖苷酶Bgl95,并对其酶学性质和应用潜力进行了综合评价。研究结果显示,Bgl95蛋白分子质量为52 kDa,最适反应温度90 ℃、最适反应pH 7.5;以对硝基苯基-β-D-吡喃葡萄糖苷为底物时,比活力达(310.5±4.1) U/mg,米氏常数(Km)为(6.1±0.5) mmol/L,最大反应速率(Vmax)为(163.9±3.6) μmol/(min·mg);Bgl95可特异性水解β-1,4糖苷键及天然底物纤维二糖、乳糖;5 mmol/L Fe2+、Mg2+及1%(质量分数)Tween 80可显著促进Bgl95的酶活力,5 mmol/L Co2+、Cu2+及1%(质量分数)SDS对酶活力有明显抑制作用;Bgl95为罕见的葡萄糖、木糖激活型β-葡萄糖苷酶,0.8 mol/L葡萄糖、1.8 mol/L木糖分别可使酶活力提升至3.55倍和4倍,即使在4 mol/L葡萄糖或木糖存在时,Bgl95仍可发挥51%和139%的酶活力。研究结果为Bgl95在纤维素降解、食品发酵等领域的应用提供理论支撑。

关键词： 嗜热玫瑰红球菌; 葡萄糖/木糖激活型; β-葡萄糖苷酶; 重组表达; 酶学性质

本文引用格式

刘洋 , 黄洁 , 张涛 , 窦康杰 , 徐有山 , 赵维萍 . 嗜热玫瑰红球菌来源葡萄糖/木糖激活型高温β-葡萄糖苷酶的重组表达与酶学性质研究[J]. 食品与发酵工业, 2026 , 52(16) : 17 -24 . DOI: 10.13995/j.cnki.11-1802/ts.046685

Abstract

To address the low catalytic activity, poor thermostability, and feedback inhibition by hydrolysis products commonly observed in β-glucosidase applications, this study identified a novel β-glucosidase Bgl95 from Thermomicrobium roseum and comprehensively evaluated its enzymatic properties and application potential.Bgl95 exhibited a molecular mass of 52 kDa, with optimal activity at 90 ℃ and pH 7.5.Using pNPG as substrate, the specific activity reached (310.5±4.1) U/mg, with a Km of (6.1±0.5) mmol/L and Vmax of (163.9±3.6) μmol/(min·mg).Bgl95 specifically hydrolyzed β-1,4-glycosidic bonds and natural substrates, including cellobiose and lactose.Enzyme activity increased significantly in the presence of 5 mmol/L Fe2+, Mg2+ and 1% Tween 80, whereas 5 mmol/L Co2+, Cu2+ and 1% SDS showed obvious inhibitory effects.Notably, Bgl95 represented a rare glucose- and xylose- stimulated β-glucosidase and 0.8 mol/L glucose and 1.8 mol/L xylose enhanced enzyme activity by 3.55-fold and 4-fold, respectively.Even in the presence of 4 mol/L glucose or xylose, Bgl95 maintained 51% and 139% of the enzyme activity.These findings provided theoretical support for the application of Bgl95 in cellulose degradation and food fermentation.

Key words： Thermomicrobium roseum; glucose- and xylose-stimulated; β-glucosidase; recombinant expression; enzymatic properties

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