产海藻糖合酶枯草芽孢杆菌破壁及转化工艺研究  

Optimization of wall breakdown and transformation technology of Bacillus subtilis producing trehalose synthase

作　　者：连战 王松江 郭传庄 隋松森 吕志飞 李小双 孙瑞成 王建彬 LIAN Zhan;WANG Songjiang;GUO Chuanzhuang;SUI Songsen;LÜZhifei;LI Xiaoshuang;SUN Ruicheng;WANG Jianbin(Zhucheng Dongxiao Biotechnology Company Limited,Weifang 261000)

机构地区：[1]诸城东晓生物科技有限公司,潍坊261000

出　　处：《中国食品添加剂》2023年第7期75-80,共6页China Food Additives

基　　金：山东省重点研发计划(重大科技创新工程)(2020CXGC010603)。

摘　　要：以细胞破碎率与海藻糖转化率为考察指标,确定发酵液破壁的最佳方式与参数。结果表明,采取高压均质机1000bar破壁两次破壁效果及海藻糖得率最佳,细胞破碎率可达99.32%,海藻糖转化率可达41.46%,较初始超声波破壁分别提升了12.31%,7.52%。采取单因素法,分别优化单酶法转化海藻糖的pH、温度,通过延长转化时间,监测转化过程中海藻糖得率的变化,确定不同转化时间糖得率的变化趋势,确定最佳的转化周期,并对胞内胞外酶转化的占比进行研究。结果显示,最佳转化条件为转化pH7.8,转化温度35℃,周期28h,在该工艺条件下进行转化,海藻糖浓度可达68.63g/L,较初始条件提升了41.88%,转化率可达48.16%,较未优化前提升了14.22%。The optimal parameters of Bacillus subtilis wall breakdown in fermentation broth were determined by wall breakdown rate and trehalose conversion rate.The results showed that when using high-pressure homogenizer at 1000 bar for twice,the yield of trehalose was the highest,cell breakage rate could reach up to 99.32%and the conversion rate of trehalose could reach 41.46%,which were increased by 12.31%and 7.52%,respectively,compared with the initial ultrasound breakdown of cell wall method.Single-factor method was adopted to optimize the pH and temperature of trehalose conversion by single-enzyme method.Change of trehalose yield during the conversion process was monitored by prolonging the conversion time.Sugar yield at different conversion times was observed to determine the best conversion cycle.The proportion of intracellular and extracellular enzyme conversion was also studied.The results showed that the optimal conversion conditions were:pH 7.8,temperature 35℃and cycle of 28 hours.Under these conditions,the concentration of trehalose could reach 68.63 g/L,which was 41.88%higher than the initial condition;the conversion rate could reach 68.63 g/L,which was 48.16%,it was 14.22%higher than the trehalose conversion rate before optimization.

关 键 词：海藻糖 破壁方式 海藻糖合酶 转化率 

分 类 号：Q815[生物学—生物工程] TS202.3[轻工技术与工程—食品科学]