高通量选育高速率产L-乳酸拟干酪乳杆菌  

A High-throughput Strategy for Screening High-L-lactic Acid-productivity Lactobacillus paracasei

作　　者：王光耀[1] 刘慧兰[1] 王永红[1] 储炬[1] 张嗣良[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点试验室,上海200237

出　　处：《食品工业》2013年第10期152-154,共3页The Food Industry

基　　金：国家重点基础研究发展计划(973计划;2013CB733600);国家重大科技专项"重大科学仪器设备开发"(2012YQ150087)

摘　　要：研究构建了一种快速初筛高产酸速率菌株的高通量方法。该方法基于溴甲酚绿在不同酸度下颜色不同的原理,使用含有溴甲酚绿的孔板培养基,实时观测培养液的颜色变化,筛选高产酸速率突变株。以拟干酪乳杆菌作为出发菌株,结合紫外诱变法和高锰酸钾-溴化钾平板复筛法,成功筛选到一株具有较高产酸速率和一定耐高糖性的菌株UV16,在含有150 g/L葡萄糖的发酵培养基中,产酸速率2.81 g/(L·h),较出发菌株提高了34.49%。本高通量初筛法操作简便,条件温和,高效快速,成本低廉,可满足大规模筛选需求。A novel high-throughput strategy for screening high-lactic acid-productivity strains was developed. The strategy typically relies upon the visual color change of bromocresol green due to the release of the lactic acid produced by strains. Incorporation of bromocresol green into the screening media allows real-time observation of the color change during fermentation. The color change can be used to distinguish high-lactic acid-productivity mutant from original strain. The strategy combined with ultraviolet （UV） and KMnO4-KBr plate, is validated for Lactobacillus paracasei. The mutant UV16 with improved L-lactic acid productivity was selected. The value of total L-lactic acid productivity increased by 34.49% was 2.81 g/（L-h） in the fermentation medium contained 150 g/L glucose. The strategy is simple, sensitive and effective and costless for high-throughput screening.

关 键 词：拟干酪乳杆菌 乳酸 高通量选育 发酵 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程]