毕赤酵母表面展示棘孢曲霉β-葡萄糖苷酶催化合成APG的工艺条件优化

生物技术通报 ›› 2014, Vol. 0 ›› Issue (6): 205-210.

毕赤酵母表面展示棘孢曲霉β-葡萄糖苷酶催化合成APG的工艺条件优化

胡霞艳, 刘端玉, 郑穗平

（华南理工大学生物科学与工程学院，广州 510006）

收稿日期:2013-12-09 出版日期:2014-06-25 发布日期:2014-06-25
作者简介:胡霞艳，女，硕士，研究方向：酶与酶应用；E-mail：lucksnail@163.com
基金资助:
国家自然科学基金项目（31171633），广东省工业科技攻关项目（2010B011000004）

Optimization of the Synthesis of Alkyl Glucoside Catalyzed by Aspergillus aculeatus β-glucosidase-displaying Pichia pastoris Whole-cells

Hu Xiayan, Liu Duanyu, Zheng Suiping

（School of Bioscience and Bioengineering，South China University of Technology，Guangzhou 510006）

Received:2013-12-09 Published:2014-06-25 Online:2014-06-25

摘要/Abstract

摘要： 使用毕赤酵母细胞表面展示棘孢曲霉的β-葡萄糖苷酶作为全细胞催化剂，以葡萄糖为底物，在水-醇双相体系中逆水解合成C6-C10烷基糖苷。讨论了含水量、葡萄糖添加量、全细胞催化剂添加量、乙酸-乙酸钠缓冲液pH值和温度等主要因素对反应的影响，并与商品酶苦杏仁β-葡萄糖苷酶、Novozym188进行比较。结果表明，Novozym188不适合逆水解合成APG，苦杏仁β-葡萄糖苷酶反应所需时间短，但最终转化率不如全细胞催化剂。在 5 mL反应体系中，合成HG的最优条件：葡萄糖0.1 g，全细胞催化剂0.05 g，10% pH 3.0 buffer；合成OG的最优条件：葡萄糖0.2 g，全细胞催化剂0.05 g，15% pH3.0 buffer；合成DG的最优条件：葡萄糖0.2 g，全细胞催化剂0.2 g，20% pH 3.0 buffer；放置55℃ 200 r/min 恒温振荡器中，反应时间为72 h，HG和DG的最大产率分别为11.69%和3.58%，而OG的产率则在96 h到达最大值6.34%。

关键词: 细胞表面展示, β-葡萄糖苷酶, 全细胞催化剂, 烷基糖苷

Abstract: Aspergillus aculeatus β-glucosidase displayed Pichia pastoris cell surface was used as whole-cell biocatalyst, glucose as substrate, catalyzed synthesis alkyl glucosides of C6-C10 with reverse hydrolysis in water-alcohol two-phase system. The effects of various reaction factors, including water content, whole-cell biocatalyst dose, glucose concentration, pH and temperature were optimized. The catalytic effect of whole-cell biocatalyst were compared with commercial enzyme almond β-glucosidase and Novozym188 . The results showed that Novozym188 was not suitable for synthesis of APG, almond β-glucosidase need shorter time than whole-cell biocatalyst, but the final conversion of whole-cell biocatalyst was higher. In 5 mL total reaction volume, the optimal reaction conditions of HG：0.1 g glucose, 0.05 g whole-cells biocatalyst, 10% pH3.0 buffer；the optimal reaction conditions of OG：0.2 g glucose, 0.05 g whole-cells biocatalyst, 15% pH3.0 buffer；the optimal reaction conditions of DG：0.2 g glucose, 0.2 g whole-cells biocatalyst, 20% pH3.0 buffer, temperature 55℃, 200 r/min. After 72 h the maximum HG and DG yield could be 11.69% and 3.58%. The highest yield of OG was 6.34% after 96 h.

Key words: Cell surface displaying, β-glucosidase, Whole-cell biocatalyst, Alkyl glucosides

胡霞艳, 刘端玉, 郑穗平. 毕赤酵母表面展示棘孢曲霉β-葡萄糖苷酶催化合成APG的工艺条件优化[J]. 生物技术通报, 2014, 0(6): 205-210.

Hu Xiayan, Liu Duanyu, Zheng Suiping. Optimization of the Synthesis of Alkyl Glucoside Catalyzed by Aspergillus aculeatus β-glucosidase-displaying Pichia pastoris Whole-cells[J]. Biotechnology Bulletin, 2014, 0(6): 205-210.

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