Productin of functional galacto-oligosccharides using transfer action of -galactosidase

利用β-半乳糖苷酶的转移作用生产功能性半乳寡糖

基本信息

批准号：
62560126
负责人：
MATSUNO Ryuichi
金额：
$ 1.34万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

项目摘要

Lactose is less soluble compared with other saccharides and tras calcium ion. These properties are the demerit in the food manufacturing and the food additive production using milk and chese whey. To add a function as bifidus factor as well as to diminish the above demerit, the preseht study aimed at the effective production of galacto-oligosaccharides using -galactosidase 1( -gal 1) and -galactosidase 2 ( -Gal 2) from Bacillus circulans which have high activity for hydrolysis of lactose and for hydrolysis of lactose and for synthesis of galactooligosaccharide, respectively. To accoplish the purpose, the following were the particular subjects.1.The enhancement of oligosaccharide formation ability of -gal 1 by chemical modification.2.Immobilization of -Gal 1 and 2 and elucidation of their properties.3.The stability of immobilyzed enzyme during continuous reaction.4.Long term continuous oligosuccharide production using either prug flow reactor or membrane reactor.5.Protection against con … More tamination and cleaning of immobilized enzyme column reactor.By treating the -Gal 1 with 0.01-3% glutaraldehyde, the amino group on the enzyme was modified from 0 to 90%. With increase in modification, increased the ability to form galacto-oligosaccharides of degree of polymerization two to five containing a glucose residue and yield of oligosaccharide from lactose reached to 40% with amino group modification 90%. Either -GAl 2 or -Gal 1 was adosorbed on to various suports and immobilized by glutaraldehyde crosslinking. Among the supports tested, Merckogel was most suitable with respect to activity and stability. however higher the specific activity, easer the immobilized enzyme deactivated reversibly. The reason for this phenomena was ascribed to the entrapment of oligosaccharides produced in the three dimensional network of crosslinked enzymes. Continuous oligosaccharide production was performed in PFR with immobilized enzyme on to merckogel SI-500(15U/g) and in membrane reactor(Diaflo cell, UF-X50) with free enzyme for 8 days without enzyme inactivation and the oligosaccharide yield from 20% lactose was maintained at 47.5%. From the adsorption-desorption experiments of milk protein, the support suitable with respect to the protection of contamination and cleaning was searched. Ion exchanger with the charge of same sign as contaminant was effective. The suggestion was obtained that the growth of micro-organism was reduced by usinglarge support with high flow rate. Less

与其他糖类和反式钙离子相比，乳糖的溶解度较低，这些特性是使用牛奶和奶酪乳清的食品制造和食品添加剂生产的缺点。预先研究旨在使用-半乳糖苷酶 1(-gal 1) 和-半乳糖苷酶 2 ( -Gal 2)来自环状芽孢杆菌，其对乳糖的水解和乳糖的水解以及低聚半乳糖的合成分别具有高活性。为了实现该目的，具体课题如下。 1.增强-的寡糖形成能力。 gal 1的化学修饰。2.-Gal 1和2的固定化及其性质的阐明。3.固定化酶在过程中的稳定性连续反应。4.使用推流反应器或膜反应器进行长期连续低聚糖生产。5.固定化酶柱反应器的污染防护和清洁。通过用 0.01-3% 戊二醛处理 -Gal 1，氨基组对酶的修饰从0%增加到90%，随着修饰的增加，形成聚合度二低聚半乳糖的能力增加。将-GAl 2 或-Gal 1 吸附到各种载体上并通过戊二醛交联固定化，从乳糖中得到的寡糖收率达到40%。就活性和稳定性而言最合适，然而比活性越高，固定化酶越容易可逆失活。交联酶的三维网络中产生的寡糖的捕获在 PFR 中使用固定化酶在 Merckogel SI-500（15U/g）上和在膜反应器（Diaflo cell，UF-X50）中进行连续寡糖生产。酶处理 8 天，没有酶失活，20% 乳糖的寡糖产量保持在47.5％，从牛奶蛋白的吸附-解吸实验中，找到了适合污染物防护和清洁的离子交换剂，表明微量污染物的生长是有效的。 -通过使用大流量和高流速来减少生物体。