褐藻胶降解菌株S10鉴定及产酶条件优化  

Identification of Alginate Degradable Strain S10 and the Optimization of Enzyme Production Conditions

作　　者：王艺欣 张健 刘芳 王共明 井月欣 矫春娜 钟静诗 邹媛婷 WANG Yi-xin;ZHANG Jian;LIU Fang;WANG Gong-ming;JING Yue-xin;JIAO Chun-na;ZHONG Jing-shi;ZOU Yuan-ting(Coll.of Food Sci.&Technol.,Shanghai Ocean Uni.,Shanghai 201306;Shandong Mar.Res.&Environ.Res.Inst.,Yantai 264006)

机构地区：[1]上海海洋大学食品学院,上海201306 [2]山东省海洋资源与环境研究院,山东烟台264006

出　　处：《微生物学杂志》2023年第1期66-76,共11页Journal of Microbiology

基　　金：山东省现代农业产业技术体系藻类产业创新团队建设项目(SDAIT-26-05);山东省现代农业产业技术体系刺参产业创;新团队建设项目(SDAIT-22-07);2020年山东省自然科学基金重点项目(ZR202011040053);。

摘　　要：探讨了褐藻胶降解菌株S10的生长条件及其对产褐藻胶降解酶活力的影响。以分离自海参肠道的褐藻胶降解菌株S10为研究对象,采用形态学观察结合16S rDNA序列分析,对菌株S10进行菌种鉴定并对其生理生化特性进行测定。以降解酶活力为指标,利用单因素、Plackett-Burman(PB)和响应面法对培养基成分和培养条件进行优化;最后对优化前后的菌株生长量、产酶活力和粗酶液稳定性进行分析。结果表明,菌株S10属于溶藻弧菌(Vibrio algindyticus);当pH 7、接种量2%(体积分数)、装液量150 mL、温度26℃、转速150 r/min、NaCl 3%(质量分数,下同)、海藻酸钠含量1.12%、硫酸铵含量0.44%、培养时间35.95 h条件下,褐藻胶降解酶活力最大(188.18 U/min)。优化后产酶活力提高30%;4℃低温更有利于该酶保存。综上,优化后的菌株S10产褐藻胶降解酶活力较高,能更好地用于降解褐藻胶,可为提高褐藻胶的利用率和进一步发掘褐藻胶寡糖的利用价值提供参考。The effects of growth conditions of strain S10 on the production activity of algin-degradable enzymes were studied.The strain S10 isolated from the intestine of sea cucumber was taken as research object and identified adopting morphological observation combined with 16S rDNA sequence analysis,and its physiological and biochemical characteristics were determined.The level of enzyme activity as the evaluation index,using single factor,Plackett-Burman(PB)and response surface methodology were conducted to optimize medium composition and culture conditions.Finally,compared the growth amount and enzyme activity of the strain S10 before and after optimization,and the stability of crude enzyme solution was analyzed.The results showed that strain S10 belonged to Vibrio algindyticus.Under the conditions of pH 7,inoculum volume 2%,liquid volume 150 mL 26℃,rpm 150 r/min,NaCl content 3%,sodium alginate content 11.20 g/L,(NH 4)2SO 4 content 4.4 g/L,and culture for 35.95 h,the highest activity of algin degradable enzyme was 188.18 U/min.The enzyme production activity of the strain was increased by 30%after optimized conditions.Low temperature of 4℃was more favorable for the preservation of the enzyme.To sum up,after the optimization,strain S10 produced higher activity of algin degradable enzyme,and could be used to degrade algin better,thus could provide references for utilization rate and further excavate the algin oligose utilization value.

关 键 词：褐藻胶降解 菌种鉴定 弧菌属 酶活力 培养条件优化 

分 类 号：Q93-331[生物学—微生物学]