新食品原料壳寡糖的酶法生产研究
目的 本论文针对高分子原料壳聚糖在食品工业应用中的难题——清洁生产与安全食用两大问题，研究了内切壳聚糖酶EC.3.2.1.132在新食品原料壳寡糖工业生产中的应用。 方法 通过广泛筛查，选育产酶活性高、性能稳定、具有单一内切模式的野生菌种。综合应用生物工程技术构建高效表达的基因重组工程菌，经优化发酵条件、建立简易纯化方法，获得了专一性内切壳聚糖酶。采用循环型清洁生产工艺用于新食品原料壳寡糖的工业化生产。 结果 从产酶量10U/mL左右的野生型曲霉菌株Jxsd-01获得成熟基因，构建重组毕赤酵母工程菌表达体系，内切壳聚糖酶蛋白产量达到0.95g/L。采用循环型清洁生产工艺酶法生产的壳寡糖含量高达98%，聚合度n = 2～10，原料转化率95%以上，生产过程中无废水、废渣产生。结论 内切壳聚糖酶应用于食品工业，实现新食品原料壳寡糖的工业化酶法生产，达到清洁、安全、高效的效果，具有应用推广的价值。
Objective To solve the problem of the applications of chitosan, which is a polymer, in food industry, we focused on two issues: the cleaner production process and the food security. The characters of endo-type chitosanase EC.3.2.1.132, and its applications were studied in the production of chitosan saccharide, a new food raw material. Methods Through the extensive screening, a strain with high enzyme activity and stable performance was got, then the endo-type chitosanase was purified . By various methods of biological engineering technology, we built a recombinant engineering yeast, which can efficiently express the aimed enzyme after the optimization of fermentation conditions, and then a simple and easy purification method was established, with a circular cleaner production process, and was applied to the industrial production of the chitosan saccharide, a new food raw material. Results The wild type Aspergillus strains Jxsd -01 could produce enzyme about 10 U/mL, and the recombinant of the gene in the engineering yeast could improve the expression system, the enzyme protein expression was 0.95 g/L. The content of the chitosan saccharide in enzymatic production was as high as 98%, with the polymers from 2 to 10. The conversion rate of raw material was more than 95%, without waste water or waste residue in the process of production. Conclusion The application of endo-type chitosanase in food industry can make the production process of the chitosan saccharide cleaner, safer and more efficient, so it has the value of wide application.
标题：新食品原料壳寡糖的酶法生产研究
英文标题：Enzymatic preparation of chitooligosaccharide, a new food raw material
作者：
游清徽 江西师范大学生命科学学院,江西省食品与生物技术产学研合作示范基地
杨萍 江西师范大学生命科学学院,江西省食品与生物技术产学研合作示范基地
徐贤柱 江西师范大学生命科学学院,江西省食品与生物技术产学研合作示范基地
裘梁 江西中医药大学生物技术研发中心,江西省食品与生物技术产学研合作示范基地
董德刚 江西中医药大学生物技术研发中心,江西省食品与生物技术产学研合作示范基地
王曼莹 江西师范大学生命科学学院,江西中医药大学生物技术研发中心,江西省食品与生物技术产学研合作示范基地
中文关键词:新食品原料,壳寡糖,壳聚糖酶,清洁生产,应用推广,
英文关键词:new food raw material,chitooligosaccharide,chitosanase,cleaner production process,application and popularization,
发表日期：2014-06-20
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