摘要
甲烷氧化菌能够以甲烷作为唯一碳源和能源物质生存.在甲烷氧化、氯代烃类污染物降解、相关化学品生产等方面具有重要的潜力.利用甲烷作为唯一碳源物质筛选分离得到一株甲烷氧化菌,并对其进行初步鉴定.在此基础上,利用单因子实验和Plackett_Burman实验等方法对获得菌株的培养条件:包括培养基条件(无机氮源种类和浓度、影响显著的金属离子及其浓度、pH值)以及培养温度进行优化.结果表明:经过16S rDNA鉴定,获得菌株与Methylocystis sp.SC2、Methylocystis hiersuta strain SV97等菌株的相似性达到99%,所以确定该菌株属于Methylocystis菌属(Ⅱ型甲烷氧化菌,甲基孢囊菌属),将本菌株命名为Methylocystis sp.M16.以1.0 g/L NH4NO3作氮源,1.0μmol/L铜离子,p H 7.00,温度30℃的条件下,甲烷去除率和菌体生物量最大.在各个因素的最优条件下,Methylocystis sp.M16菌液吸光值(A600 nm)均在0.5以上,甲烷去除率在95%以上.本研究分离得到一株Ⅱ型甲烷氧化菌Methylocystis sp.M16,并获得优化的培养基和培养条件,有望为M16菌应用提供理论参考.
Methanotroph can survive with methane as the sole carbon and energy source. With enzyme system of methane oxidation, Methanotroph play an important role in mitigation of methane and degradation of pollutant, such as chlorinated hydrocarbon, and chemical production. In this study, methane was used as the sole carbon source to screen and separate a methanotroph and it was primarily identified. Molecular biological methods （16S rDNA gene to be sequenced and aligned on Online database NCBI） were used to identify the methanotroph. The best type of inorganic nitrogen source and its proper concentration was optimized by single factor experiment. The most significant trace elements were chosen by using Plackett- Burman method. The concentration of most significant trace elements, the proper pH and temperature were also optimized by single factor experiment. The methanotroph obtained from soil was identified as Methylocystis and named Methylocystis sp. M16. The optimized conditions for methane removal and growth ofMethylocystis sp. M16 were NH4NO3 of 1.0 g/L, Copperion of 1.0μmol/L, pH of 7.0 and temperature of 30 ℃. Under the optimal condition, the absorption values （A600nm of Methylocystis sp. M16 were above 0.5 and methane removal efficiency above 95%. The results could provide theoretical reference for the application ofMethvlocvstis Sla. M16.
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2015年第3期455-463,共9页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金项目(41275161)
石河子大学青年基金项目(2012RCZXJQ05)资助~~
关键词
甲烷氧化菌
甲基孢囊菌属
菌种筛选
培养条件优化
methanotroph
Methylocystis
strain screening
optimization of culture condition
