摘要
为提高阿卡波糖发酵水平,制备了阿卡波糖产生菌犹他游动放线菌ZJB-08196的原生质体并对原生质体进行诱变处理。考察了甘氨酸的添加方式及浓度、菌龄、酶浓度、酶解时间、酶解温度、菌体浓度对原生质体制备和再生的影响,并对再生培养基进行了优化。较佳的原生质体制备与再生条件为:菌体一级培养56 h后,转接入含0.4%甘氨酸的菌丝培养基中二级培养22 h,收集菌体并调整菌体浓度为0.2 g/mL,用10 mg/mL溶菌酶35℃水浴酶解4 h后,涂布添加4 g/L L-脯氨酸和0.2 g/L聚乙烯吡咯烷酮的R2YE固体平板,原生质体的再生率达到了15.14%。原生质体经紫外诱变处理,筛选到1株高产突变株UN-52,阿卡波糖产量达到5 165.2 mg/L,比出发菌株提高了12%。
In order to improve the production of acarbose,protoplasts of an acarbose-producing strain Actinoplanes utahensis ZJB-08196 were prepared and treated with ultraviolet irradiation.The effects of factors including glycine concentration,mycelium age,lysozyme concentration,treating time,temperature and mycelial concentration on protoplast formation and regeneration were investigated.Moreover,the effect of the addition of L-proline,bovine serum albumin（BSA）,polyvinylpyrrolidone（PVP） into regeneration medium on protoplast regeneration was further examined.The results indicated that protoplasts preparation and regeneration were achieved with following method.The 56 h-old seeds were inoculated into mycelia medium with 0.4% glycine and cultured for 22 h.Then the mycelia were collected and treated by 10 mg/mL lysozyme at 35 ℃ for 4 h and then spread on the R2YE-agar（regeneration medium supplemented with L-proline 4 g/L and PVP 0.2 g/L） plates.The maximum rate of protoplast regeneration reached 15.14%.After ultraviolet irradiation,a high-yield mutant UN-52 was obtained.Acarbose production of 5 165.2 mg/L with mutant UN-52 was obtained,increased by 12% compared with the original strain.
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2013年第5期37-43,共7页
Food and Fermentation Industries
基金
浙江省重大科技专项(No. 2008C03004-1)
