表面显示的曼诺分析和奇丁分析酶使用丁糖甘结合lysM域
Hoang-Minh Nguyen1, Khanh-Trang V Le2,3, Ngoc-Luong Nguyen4
1Department of Biotechnology, Faculty of Chemical Engineering, The University of Da Nang─University of Science and Technology, 54 Nguyen Luong Bang, Da Nang 550000, Vietnam.
Journal of agricultural and food chemistry
|May 22, 2024
概括
这项研究开发了使用 Lactiplantibacillus plantarum 的非转基因全细胞生物催化剂,以显示基托酶和曼南酶酶. 双 LysM 域实现了高的固定产量,使得预微生物寡糖的生产.
科学领域:
- 生物技术是生物技术.
- 微生物工程 微生物工程
- 酶技术 酶技术是一种
背景情况:
- 乳植物菌 (Lactiplantibacillus plantarum) 是一种适用于全细胞生物催化剂开发的食品级细菌.
- 在细菌表面的酶固定增强了生物催化剂的稳定性和可重复使用性.
- LysM 域是细菌中蛋白质表面显示的有效.
研究的目的:
- 评估单个和双个LysM域对化酶 (CsnA) 和曼南酶 (ManB) 在L. plantarum上的有效性.
- 为了优化L. plantarum上显示的CsnA和ManB的固定产量和酶活性.
- 评估这些工程细胞生产益生菌寡糖的潜力.
主要方法:
- 编码CsnA和ManB的基因与LysM域融合,并在大肠杆菌中得到表达.
- 再组合蛋白质被固定在L. plantarum WCFS1.1的表面上.
- 使用西方斑点和流细胞计验证了表面定位.
- 酶活性被量化,并通过高性能离子交换色谱分析产品的形成.
主要成果:
- 在L. plantarum上成功地显示了CsnA和ManB的表面显示.
- 来自Lp_2162的双 LysM 域实现了最高的固定收益率 (44-48%).
- 最佳的酶活性为812U/g的曼南酶和508U/g的基托桑酶被记录.
- 证明了曼诺-寡糖和基托-寡糖的产生.
结论:
- 显示CsnA和ManB的非转基因L. plantarum细胞是有效的全细胞生物催化剂.
- 双 LysM 域是 L. plantarum 上酶表面显示的高级.
- 改造的L. plantarum具有可持续生产益生菌寡糖的潜力.
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