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Production of novel oligosaccharides by microbes and evaluation of their function

微生物生产新型低聚糖及其功能评价

基本信息

批准号：
07556087
负责人：
TOMITA Fusao
金额：
$ 6.72万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

1.Preparation of Raw Material for Oligosaccharides Production : Extraction and purification of inulin from chicory was attempted according to the method of sugar purification from beet, and was successfully achieved. Preparation of levan was also carried out by using levan producing bacteria, and levan was recovered and purified from its culture broth. More simple purification process for preparation of raw material has attempted by such method as addition of oligosaccharide-producing enzyme to crude extracts.2.Screening and Improvement of Oligosaccharide Producing Microbe : Oligosaccharide producing microbe was screened from plant in addition to soil as a screening source, and xylobiose-producing fungi was newly isolated. Molecular cloning of ift and lft gene, which encode DFAIII and DFAIV producing enzyme respectively, were executed. Expression systems of ift and lft gene was constructed in E.coli system, and higher activities were gained by using these system rather these original strains.3.Establishment of Oligosaccharide Production Process : Culture conditions for oligosaccharides producing enzymes and reaction conditions were optimized by characterizing microbes and enzymes producing DFAIII,DFAIV,inulotriose and levanbiose. Large scale preparations of purified DFAIII and DFAIV were gained by the process such as fermentation of reaction mixturi by yeast, ion exchange column and crystallization.4.Evaluation of Physiological Function of Oligosaccharide : Properties of DFAIII with respect to both industrial processing and nutritional function by using microbe, rat and human were examined. In the case of rat, calcium absorption was increased by adding DFAIII to feed, and the regions of absorption were determined in large and small intestines. Physiological fuction of DFAIV has also been evaluation.

1.低聚糖生产原料的制备：采用甜菜糖提纯的方法，对菊苣菊粉的提取纯化进行了尝试，成功地实现了菊苣菊糖的提取纯化。利用利凡产生菌进行了利凡的制备，并从其发酵液中回收纯化了利凡。2.低聚糖产生菌的筛选与改良：以植物和土壤中的低聚糖产生菌为筛选来源，从植物中筛选出低聚糖产生菌，并新分离到木二糖产生菌。对DFAIII和DFAIV产生酶基因ift和lft进行了分子克隆。3.低聚糖生产工艺的建立：通过对产DFAIII、DFAIV、菊粉三糖和左旋二糖的微生物和酶的特性研究，优化了低聚糖产酶的培养条件和反应条件。通过酵母菌发酵、离子交换柱层析、结晶等步骤，获得了DFAIII和DFAIV的规模化制备。4.低聚糖生理功能的评价：利用微生物、大鼠和人体对DFAIII进行了工业加工和营养功能的研究。在大鼠饲料中添加DFAIII可增加钙的吸收，并测定了大肠和小肠的吸收区域。并对DFAIV的生理功能进行了评价。