黑曲霉YY-22产酸性果胶酶的分离纯化

Purification of Acidic Pectinases Produced by Aspergillus niger YY-22

摘要:

本文研究了从黑曲霉YY-22发酵产酸性果胶酶粗酶液中分离出较高纯度果胶裂解酶（PL）、聚半乳糖醛酸酶（PG）、果胶酯酶（PE）的方法，同时考察了主要组分PL在各分离阶段的纯化效果。依次采用硫酸铵盐析、疏水相互作用层析及离子交换层析对果胶酶PG、PE、PL进行分离。结果表明：果胶酶粗酶液中硫酸铵饱和浓度为65%时，沉淀中PL回收率最大，部分杂蛋白在盐析过程中被分离出来；沉淀复溶后经Phenyl-Sepharose FF疏水相互作用层析首先分离出PE活性组分，又经Q-Sepharose HP强阴离子交换层析分别得到PG和PL活性组分。果胶酶粗酶液中主要组分PL经三步纯化后的比活力达到79.37 U/mg蛋白，纯化倍数为13.30，活力回收为33.05%。较高纯度PL、PG、PE的获得，为进一步研究酶的基本性质及其在果蔬加工和葡萄酒中的应用奠定基础。

Abstract:

Purification of pectin lyase (PL), polygalacturonase (PG), and pectin esterase (PE) from the fermentation culture supernatant of Aspergillus niger YY-22 was investigated. All the three enzymes PL, PG and PE were precipitated by ammonium sulfate precipitation (0~65%). PE was further purified by hydrophobic interaction chromatography using Phenyl-Sepharose FF, while PG and PL were purified by Q-Sepharose HP ion exchange chromatography. The highest recovery of PL was obtained with 65% ammonium sulfate saturation precipitation. After three steps of separation, PL was purified 13.30 folds with a recovery of 33.05% and a specific activity of 79.37 U/mg.