摘要
[目的]从海水中获得高产几丁质酶菌株并优化培养条件。[方法]采集湛江市金沙湾海水作为样品,以胶体几丁质为唯一碳源筛选分离产几丁质酶菌株,DNS法测定酶活力并进行分子鉴定;单因素试验和正交优化其产酶最佳条件。[结果]初筛得到6株能降解几丁质的细菌,其中编号为Z-4的菌株几丁质酶活力最强,形态鉴别及16S rDNA序列分析鉴定为嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia)。菌株Z-4优化后的最佳产酶条件为可溶性淀粉添加量1.5%、蛋白胨添加量2.5%、KH2PO4添加量0.04%,在30℃、pH 7.0、接种量2%、160 r/min、培养72 h,酶活力达到0.452 U/mL,相比优化前提高了5倍。[结论]研究结果为海洋副产物有效利用几丁质改善环境提供重要参考。
[Objective]To obtain high⁃yield chitinase strains from seawater and optimize culture conditions.[Method]The seawater from Jin⁃sha Bay of Zhanjiang City was collected as samples,chitinase producing strains were isolated using colloidal chitin as the only carbon source,the enzyme activity was measured by DNS method and molecular identification;single factor experiment and orthogonal optimization were used to optimize the optimal conditions for enzyme production.[Result]Six strains of chitinase⁃degrading bacteria were screened.The strain with the strongest chitinase activity was Z⁃4.The strain was identified as Stenotrophomonas maltophilia by morphological observation and 16S rDNA se⁃quence analysis.The optimum conditions for producing enzyme of strain Z⁃4 were obtained as follows:soluble starch addition of 1.5%,peptone addition of 2.5%,KH2 PO4 addition of 0.04%;at 30℃,pH 7.0,inoculation amount of 2%,160 r/min and 72 hours of cultivation,the enzyme activity reached 0.452 U/mL,which was 5 times higher than before optimization.[Conclusion]The research results provide an important refer⁃ence for the effective use of chitin in the ocean to improve the environment.
作者
祁红兵
张敏怡
QI Hong-bing;ZHANG Min-yi(School of Life Science and Technology,Lingnan Normal University,Zhanjiang,Guangdong 524048)
出处
《安徽农业科学》
CAS
2024年第16期1-5,共5页
Journal of Anhui Agricultural Sciences
基金
湛江市非资助科技攻关计划项目(2023B01023)
岭南师范学院校级教育教学改革项目(岭师教务〔2022〕154号)。
关键词
海水
细菌
几丁质酶
培养条件优化
筛选鉴定
Seawater
Bacteria
Chitinase
Optimization of culture conditions
Screening and identification
