热稳定k-卡拉胶酶产生菌Bacillus sp.Car19的发酵条件优化  

Optimization of fermentation conditions for thermostable κ-carrageenase production of strain Bacillus sp. Car19

作　　者：谢买胜 李江[1] 林学政[1] 何培青[1] XIE Mai-sheng LI Jiang LIN Xue-zheng HE Pei-qing(First Institute of Oceanography, SOA, Qingdao 266061, China)

机构地区：[1]国家海洋局第一海洋研究所,山东青岛266061

出　　处：《海洋科学》2016年第11期7-16,共10页Marine Sciences

基　　金：海洋公益性行业科研专项项目(201505026);青岛市应用基础研究计划项目(14-2-4-14-jch);国家海洋局海洋生物活性物质与现代分析技术重点实验室开放基金(MBSMAT-2015-06)~~

摘　　要：对一株分泌热稳定κ-卡拉胶酶印尼热泉菌进行了种属鉴定,并采用响应面法对该菌发酵产酶条件进行了优化。鉴定结果表明,该菌株属于芽孢杆菌属(Bacillus),命名为Bacillus sp.Car19(Gene Bank:KT865196)。发酵条件优化结果显示,9个环境因子影响Bacillus sp.Car19产酶量。其中影响Bacillus sp.Car19产酶量的三个主要因素分别为培养温度、培养基中Cu^(2+)浓度和培养基中Na Cl浓度。综合次要因素对Bacillus sp.Car19产酶影响,Bacillus sp.Car19最佳产酶发酵条件为:培养温度52.31℃、Cu^(2+)浓度6.93 mmol/L、Na Cl浓度37.03 g/L,培养基p H为6,接种量1%,培养时间36 h,半乳糖浓度0.3 g/L,硝酸铵浓度7 g/L,卡拉胶浓度0.5 g/L。优化后发酵上清液酶活力达到15.21 U/m L,与优化前相比提高了1.5倍。In this study, we identified a high-yield K-carrageenase bacteria strain by its 16S rDNA sequence and optimized the fermentation conditions using a response surface methodology. The results show that the hot-spring bacterium was 99% identical to Bacillus sp. and therefore we assigned it to the Bacillus genus and named it Bacillus sp. Carl9 （GenBank accession number： KT865196）. The optimized fermentation conditions of Bacillus sp. Carl9 involve nine factors that affect Bacillus sp. Carl9 carrageenase production, and the three main factors are culture temperature, Cu2＋ concentration, and NaC1 concentration. Combined with subordinate factors, the final optimized fermentation conditions for maximal activity, as determined by regression analysis are as follows： a culture temperature of 52.31℃, a Cu2＋ concentration of 6.93 mmol/L, an NaCI concentration of 37.03 g/L, medium pH = 6, 1% inoculation quantity, 36 h incubation time, a galactose concentration of 0.3 g/L, an ammonium nitrate concentration of 7 g/L, and a carrageenan concentration of 0.5 g/L. The supernatant carrageenase activity after optimized fermentation can reach 15.82 U/mL, which is 1.5 times that without optimization.

关 键 词：热泉菌 卡拉胶酶 优化 响应面 

分 类 号：Q178.53[生物学—水生生物学]