食品与发酵工业 >

2026 , Vol. 52 >Issue 3: 148 - 157

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.043119

研究报告

酵母孢子固定化木聚糖酶及其酶学性质研究

  • 邹延 ,
  • 杨书恒 ,
  • 张京涛 ,
  • 曾雪莹 ,
  • 潘聪 ,
  • 周芳 ,
  • 杨宗玲 ,
  • 段盛林 ,
  • 刘国玉 ,
  • 苑鹏
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  • (中国食品发酵工业研究院有限公司,功能主食创制与慢病营养干预北京市重点实验室,北京,100015)
第一作者:硕士研究生(刘国玉工程师和苑鹏教授级高级工程师为共同通信作者,Email:lgy17091037820@163.com;Ypt86@163.com)

收稿日期: 2025-04-25

  修回日期: 2025-06-04

  网络出版日期: 2026-03-13

基金资助

北京市科技计划项目(Z231100003723014)

收起

Enzymatic properties of xylanase immobilized by yeast spores

  • ZOU Yan ,
  • YANG Shuheng ,
  • ZHANG Jingtao ,
  • ZENG Xueying ,
  • PAN Cong ,
  • ZHOU Fang ,
  • YANG Zongling ,
  • DUAN Shenglin ,
  • LIU Guoyu ,
  • YUAN Peng
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  • (China National Research Institute of Food and Fermentation Industries Co.Ltd., Beijing Key Laboratory of the Innovative Development of Functional Staple and Nutritional Intervention for Chronic Diseases, Beijing 100015, China)

Received date: 2025-04-25

  Revised date: 2025-06-04

  Online published: 2026-03-13

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摘要

木聚糖酶作为半纤维素降解的关键酶,在生物质转化、食品加工及造纸领域具有重要的应用价值。然而,游离态木聚糖酶普遍存在热稳定性差、pH耐受范围窄、无法回收使用等问题,严重制约其工业化应用。针对这一瓶颈,该研究基于酿酒酵母孢子壁的独特结构,将黑曲霉来源的木聚糖酶XynB固定于孢子壁上,通过载体空间位阻效应和静电吸附作用提升酶学稳定性。研究首先通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE)与荧光分析确认XynB可在孢子中表达并定位于孢子壁上。其次,以野生型AN120、孢子壁缺陷型菌株osw2Δ及dit1Δ为宿主,系统评估其孢子固定化XynB的酶学性质。结果表明,osw2Δ孢子固定的XynB表现出最优酶学特性:最适反应温度为40 ℃,最适pH为5.0;与游离酶相比,该固定化酶在80 ℃孵育120 min后仍保持30%的残留酶活力,在pH 4.0~8.0范围内孵育120 min后残留酶活力超过50%,pH耐受范围显著扩大。此外,osw2Δ孢子固定化XynB对金属离子、EDTA、去污剂(Triton X-100、SDS)及高盐环境均表现出显著耐受性,经5次循环催化后仍保留56%的初始酶活力,且对木聚糖类底物具有高度特异性。该研究是酿酒酵母孢子固定化半纤维素酶的新尝试,在木聚糖水解及木质纤维素生物质转化领域展现出广阔的应用前景。

关键词： 酿酒酵母; 孢子固定化; 木聚糖酶; 酶学性质

本文引用格式

邹延 , 杨书恒 , 张京涛 , 曾雪莹 , 潘聪 , 周芳 , 杨宗玲 , 段盛林 , 刘国玉 , 苑鹏 . 酵母孢子固定化木聚糖酶及其酶学性质研究[J]. 食品与发酵工业, 2026 , 52(3) : 148 -157 . DOI: 10.13995/j.cnki.11-1802/ts.043119

Abstract

Xylanase, a key enzyme involved in hemicellulose degradation, plays a pivotal role in biomass conversion, food processing, and papermaking industries.However, the industrial application of free xylanase is often restricted due to its poor thermal stability, narrow pH adaptability, and limited reusability.To address these limitations, a novel immobilization strategy was developed by anchoring Aspergillus niger-derived xylanase XynB onto the spore surface of Saccharomyces cerevisiae, leveraging the unique structural characteristics of yeast spores.The immobilized enzyme exhibited enhanced stability through spatial hindrance and electrostatic interactions with the spore carrier.The surface expression and localization of XynB on spores were verified by SDS-PAGE and fluorescence microscopy.Enzymatic properties were systematically investigated using wild-type AN120 and spore wall-deficient mutants osw2Δ and dit1Δ.Among them, the osw2Δ-spore-displayed XynB exhibited superior catalytic performance, with optimal activity at 40 ℃ and pH 5.0.Compared with the free enzyme, the immobilized form retained about 30% of its initial activity after incubation at 80 ℃ for 120 min, and >50% activity was maintained after 120 min within the pH range of 4.0-8.0, indicating significantly improved thermostability and pH tolerance.Moreover, the immobilized enzyme showed excellent resistance to metal ions, EDTA, detergents (Triton X-100, SDS), and high salinity.After five reuse cycles, 56% of the initial activity was retained.Notably, the immobilized XynB demonstrated high substrate specificity, particularly toward birch xylan.These findings provide a promising strategy for xylanase immobilization and offer valuable insights for its application in xylan hydrolysis and lignocellulosic biomass conversion.

Key words： Saccharomyces cerevisiae; spore immobilization; xylanase; enzymatic property

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