黑曲霉脂肪酶全细胞生物催化剂的制备及其实用性质。

Preparation of a whole-cell biocatalyst of Aspergillus niger lipase and its practical properties.

机构信息

Key Laboratory of Molecular Biophysics, Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.

出版信息

J Agric Food Chem. 2010 Oct 13;58(19):10426-30. doi: 10.1021/jf1008555.

DOI:10.1021/jf1008555

PMID:20828152

Abstract

Aspergillus niger lipase (ANL), a widely used hydrolase, was displayed for the first time on the surface of Saccharomyces cerevisiae using a-agglutinin as an anchor protein. Localization of ANL on the cell surface was confirmed by immunofluorescence microscopy. The displayed ANL was confirmed to be active toward tributyrin and p-nitrophenyl caprylate (pNPC). The hydrolytic activity toward pNPC reached 43.8 U/g of dry cell weight after induction by galactose for 72 h. The ANL-displaying cells were characterized for their use as whole-cell biocatalysts. The optimum temperature was 45 °C, and the pH was 7.0. The cells had good thermostability, retaining almost 80% of the full activity after incubation at 60 °C for 1 h, and >80% of the full activity at 50 °C for 6 h. The displayed lipase showed a preference for medium-chain fatty acid p-nitrophenyl esters. Therefore, the produced whole-cell catalyst is likely to have a wide range of applications.

摘要

黑曲霉脂肪酶（ANL）是一种广泛应用的水解酶，首次通过凝集素作为锚定蛋白在酿酒酵母表面进行展示。通过免疫荧光显微镜确认了 ANL 在细胞表面的定位。展示的 ANL 被证实对三丁酸甘油酯和对硝基苯辛酸酯（pNPC）具有活性。经半乳糖诱导 72 h 后，对 pNPC 的水解活性达到 43.8 U/g 干细胞重量。对展示 ANL 的细胞进行了全细胞生物催化剂的特性表征。最适温度为 45°C，pH 值为 7.0。细胞具有良好的热稳定性，在 60°C 孵育 1 小时后保留几乎 80%的全部活性，在 50°C 孵育 6 小时后保留>80%的全部活性。展示的脂肪酶对中链脂肪酸对硝基苯酯表现出偏好。因此，所产生的全细胞催化剂可能具有广泛的应用范围。