高产胞外多糖乳酸菌菌株的选育及其在酸奶生产中的应用-食品科学专业论文.docxVIP

摘要摘 摘要 摘 要 酸奶中常出现凝胶易断裂、粘稠度低、乳清易于析出等质量问题，使用产粘(胞外 多糖)乳酸菌可以大大降低酸奶的凝胶脱水收缩。所产生的胞外多糖对酸奶的流变学特 性和组织状态有益。本实验从16株乳酸菌中筛选出一株高产粘菌，使用诱变育种技术对 其生产性能进行改良，然后应用于酸奶生产中，对乳糖利用率、乳清析出量、粘度及最 终产品的可接受性进行系统的研究。 (从kefir中分离到lO株乳酸菌与实验室保存的6株乳酸菌分别于最适条件下发酵， 用三氯乙酸(TCA)去除蛋白、酒精沉淀多糖后，采用苯酚．硫酸法测多糖(以葡萄糖为 标准)。以胞外多糖产量和发酵液粘度为指标，筛选出一株产粘7号菌，其产EPS量为 ／ 265．93rag／L。f 通过平板初筛筛选出7-12作为出发菌株，以紫外线(UV)、硫酸二乙酯(DES)、 亚硝基胍(NTG)作为诱变剂，单一或复合诱变连续处理6代。从2000多个菌株中通 过初筛和多次复筛，选育出高产7F379变株，其产胞外多糖量达584 5mg／L，比出发菌 株提高了129．4％。连续传代6次后其产EPS水平呈现不稳定趋势。从选育谱系分析单 因子以NTG的诱变效应最佳，DES次之，uv较差。单一诱变剂的最适剂量为：uv照 射3min；NTG 300 u g／mL处理40rain；DES 1 u L(0 2M)／u L原菌液。 将产粘菌株与普通酸奶生产菌结合用于制作酸奶，用链霉蛋白酶在DH 7 5，30℃条 件下消化16h，再用95％乙醇沉淀、离心、透析获得粗多糖，HPLC法测定发酵液中的 糖组成及EPS的单体组成。研究表明：诱变后的7F379菌株乳糖利用速度及半乳糖合成 速度都较快，培养24h后大约有一半的乳糖被利用，在整个培养周期尤其是培养10．5h 后半乳糖产生速度明显增加。产粘7号菌在诱变前后与不产粘嗜热链球菌共同发酵产生 的EPS单糖组成不变，都是由葡萄糖、半乳糖和鼠李糖组成；而不产粘嗜热链球菌与不 产粘保加利亚乳杆菌产生的EPS不含鼠李糖，只有葡萄糖和半乳糖。诱变后的产粘菌与 不产粘嗜热链球菌组合生产酸奶乳清析出最少，通过感官评定认为：有产粘菌参与形成 的发酵乳总体可接受性好。 关键词乳酸菌：胞外多耱；诱变；酸奶 Screening Screening of Productive ExopOlysaccharide Lactic Acid Bacteria and Application in Yogurt Production Abstract Yogurt manufacturers are often faced with fracture of the gel，low。viscosity and wheying-off．Using polymer producing strains of lactic acid bacteria can reduce considerably the syneresis of yogurt．The appearance of EPS is beneficial to the rheological and texture properties of yogurt．In this study one high ropy strain was selected from 16 lactobacillus， then the mutation techniques improved in order to modify its producing properties and to make yogurt，lactose utilization ratio，wheying off,viscosity and acceptability of the ultimate products were investigated systematically． Six lactobacillus preserved in the laboratory∞d ten isolated from kefir grains were fermented in the optimum conditions．TCA and ethanol were used to precipate protein and EPS respectively．Total sugar concentration was determined by the phenol-sulfuric method using glucose as a standard．EPS quantity and cultured broth viscosity were parameters，strain 7 was s