解脂耶氏酵母细胞表面展示乳糖水解酶高效水解乳糖 <sup>*</sup>

doi:10.13523/j.cb.20180806

中国生物工程杂志

2018, Vol. 38

Issue (8): 41-49 DOI: 10.13523/j.cb.20180806

技术与方法

解脂耶氏酵母细胞表面展示乳糖水解酶高效水解乳糖 ^*

王艺颖,程海荣(

)

上海交通大学生命科学技术学院微生物代谢国家重点实验室上海 200240

Cell Surface-Displaying the Lactose Hydrolase on Yarrowia lipolytica: a New Approach to Lactose Hydrolysis

Yi-ying WANG,Hai-rong CHENG(

)

School of Life Science and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, China

全文: PDF(1633 KB) HTML

摘要：

乳糖是婴幼儿获取能量的重要碳源之一,但乳糖需要在乳糖酶的作用下水解成半乳糖与葡萄糖后才能被吸收。缺少乳糖分解酶的婴幼儿在摄入含乳糖的食品后,未被消化的乳糖会直接进入大肠,刺激大肠蠕动加快,造成一系列不适应症状即乳糖不耐症,我国属于乳糖不耐症高发国家。因此,解决乳糖的体外水解问题对减轻该症状有重要的意义。研究通过将β-半乳糖苷酶(也称为乳糖水解酶)表面展示在食品安全微生物解脂耶氏酵母(Yarrowia lipolytica)细胞表面,通过培养获得该酵母,然后直接利用酵母细胞来水解乳糖生成半乳糖与葡萄糖。采用该工程酵母细胞(HCY10),能在24小时内完全水解50g/L的乳糖,生成半乳糖与葡萄糖。该方法具有高效、简便的优点,能为乳糖的高效水解提供一条新的途径。

关键词： 乳糖; β-半乳糖苷酶; 乳糖水解酶; 解脂耶氏酵母; 酵母表面展示技术

Abstract:

Lactose is one of the important carbon sources for infants to obtain energy, but lactose needs to be hydrolyzed by enzyme to galactose and glucose before being absorbed. Infants and young children lacking lactose hydrolase can fall in lactose intolerance when consuming lactose-containing food. Lactose intolerance is a symptom of high incidence in China. Therefore, lactose hydrolysis with high efficiency in vitro is of great importance to alleviate the symptoms. The lactose hydrolase (also known as β-galactosidase) was displayed on the cell surface of the food-satefy-grade yeast Yarrowia lipolytica, and the engineered yeast cells are used directly to hydrolyze lactose to produce galactose and glucose. The engineered yeast cell (HCY10) can completely hydrolyze 50g/L lactose in 24 hours to produce galactose and glucose. The method has the advantages of high efficiency and simplicity, and it can provide an alternative solution for the efficient lactose hydrolysis in vitro.

Key words: Lactose β-galactosidase; Lactose hydrolase Yarrowia lipolytica Yeast surface display technology

收稿日期: 2018-03-08 出版日期: 2018-09-11

ZTFLH:

Q819

基金资助: 国家自然科学基金(21661140002)

通讯作者: 程海荣 E-mail: chrqrq@sjtu.edu.cn

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引用本文:

王艺颖,程海荣. 解脂耶氏酵母细胞表面展示乳糖水解酶高效水解乳糖 ^*[J]. 中国生物工程杂志, 2018, 38(8): 41-49.

Yi-ying WANG,Hai-rong CHENG. Cell Surface-Displaying the Lactose Hydrolase on Yarrowia lipolytica: a New Approach to Lactose Hydrolysis. China Biotechnology, 2018, 38(8): 41-49.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20180806 或 https://manu60.magtech.com.cn/biotech/CN/Y2018/V38/I8/41

图1 含来自米曲霉112菌株乳糖水解酶基因的表面展示质粒

图2 在YNLHX平板上生长的酵母转化子

图3 Y.lipolytica转化子的分子鉴定

图4 不同的酵母转化子转化5%乳糖的HPLC 检测结果

图5 20个深蓝色转化子乳糖水解酶活性比较

图6 HCY10转化子水解不同浓度乳糖的效率

图7 HCY10转化子与对照Y.lipolytica CGMCC7326菌株合成赤藓糖醇的性能比较

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