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一株海藻多糖降解菌的分离鉴定及产酶条件优化
朱大玲1, 唐啸龙2, 张宝玉3, 杨振芳2, 王乐普1, 秦乐宁1, 张 蕾1
1.天津市海洋化学与资源重点实验室, 天津科技大学 化工与材料学院;2.天津科技大学 海洋与环境学院;3.中国科学院 海洋研究所
摘要:
从大型褐藻藻体分离获得一株具有高效降解琼胶和褐藻胶能力的革兰氏阴性菌菌株ST-6。16SrDNA序列分析结果表明, 该菌株与海绵假单胞菌的相似度达到99%, NJ法构建系统进化树也与海绵假单胞菌归为一类, 鉴定为海绵假单胞菌Pseudomonas pachastrellae ST-6。在2216E培养基、30℃培养条件下, 海绵假单胞菌ST-6的生长曲线表明, 接种10~48 h为菌株的指数生长期, 48~72 h为生长稳定期。产酶结果表明, 菌株ST-6在指数生长期时菌液的胞外琼胶酶相对酶活力较高, 在接种48 h时, 菌液的琼胶酶相对酶活力最高为249.15 U/mL。此外, 发现菌株ST-6的琼胶酶和褐藻胶酶的分泌类型分别为非诱导型和诱导型。采用单因素分析法对其生长和产酶条件进行分析, 结果表明, 海绵假单胞菌ST-6最适生长条件为: 温度25~35℃, pH值5~9; 最适产琼胶酶条件为: 温度30℃, pH值为7, 琼胶浓度0.3%。最适产褐藻胶酶条件为: 温度35℃, pH值为9。在温度30℃、pH9和褐藻酸钠浓度0.15%的培养条件下, 海绵假单胞菌ST-6获得最高胞外褐藻胶酶相对酶活力为135.54 U/mL。
关键词:  海绵假单胞菌  琼胶酶酶活  褐藻胶酶酶活  3,5-二硝基水杨酸比色法  产酶条件优化
DOI:10.11759/hykx20161222001
分类号:
基金项目:青岛海洋科学与技术国家实验室开放基金项目(0F2015N015); 天津科技大学青年教师创新基金项目(2015LG10); 天津市科技支撑计划项目(15ZXCXSF00040); 国家自然科学基金项目(21406169)
Isolation identification and enzyme-producing conditions analysis of a seaweed polysaccharides-degrading bacteria
ZHU Da-ling,TANG Xiao-long,ZHANG Bao-yu,YANG Zhen-fang,WANG Le-pu,QIN Le-ning,ZHANG Lei
Abstract:
In this study, we isolated a marine agar and alginate-degrading Gram-negative bacterial strain ST-6 from brown seaweed. The results of our 16S rDNA sequence analysis indicate that the similarity was 99% between the strain ST-6 and the strains of Pseudomonas pachastrellae. We classified these strains as a class in phylogenetic trees using the NJ method and identified the strain as P. pachastrellae ST-6. After culturing in a 2216E medium at 30℃, we simultaneously analyzed the biomass accumulation and enzyme production of P. pachastrellae ST-6. The growth curve results indicate the exponential and stationary growth phases to be 10–48 h and 48–72 h after inoculation, respectively. The enzyme production results indicate that the enzyme activity was higher in the exponential growth phase and that the highest extracelluar agarase relative activity was 249.15 U/mL 48 h after inoculation. In addition, the secretion types of agarase and alginate lyase of P. pachastrellae ST-6 were non-inducible and inducible, respectively. The optimum growth conditions were a culture temperature of 25–35℃ and initial pH values of 5 and 9, respectively. The optimum agarase-producing conditions were a culture temperature of 30℃, an initial pH value of 7, and an initial agar concentration of 0.30%. The optimum alginate lyase-producing conditions were a culture temperature of 35℃, an initial pH value of 9, and an alginate concentration of 0.30%. We obtained a maximum relative extracelluar alginate lyase activity of 135.54 U/mL in culture conditions with a temperature of 30℃, a pH value of 7, and an alginate concentration of 0.30%.
Key words:  Pseudomonas pachastrellae  agarase activity  alginate lyase activity  3, 5-Dinitrosalicylic acid colorimetry method  condition optimization of enzyme production.
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