摘 要：本论文主要对甲烷氧化菌B22培养条件、生长情况及其在甲醇、乙醇底物中代谢产物进行初步分析，以考察该菌在甲烷转化中的作用。论文首先进行了甲烷氧化菌B22培养条件的初步优化，结果发现：1）甲烷氧化菌B22的最佳培养氮源是硝酸钾；2）最适生长温度是30 ℃；3）最适生长pH是7；4）向培养基中添加维生素复合物对甲烷氧化菌B22的生长起促进作用。其次，对甲烷氧化菌B22在甲醇、乙醇为底物的生长曲线及代谢产物进行初步研究，结果发现：B22在有氧环境下的的生长速度和繁殖总量均高于无氧环境；该菌在以甲醇和乙醇为底物进行有氧发酵时分别产生了甲酸和乙酸，而在厌氧发酵时只产生了乙酸没有甲酸。本论文的研究将为培养条件进一步优化及产物分析提供依据。65511

毕业论文关键词：甲烷氧化菌，甲醇，乙醇，液相色谱

Abstract: This paper studied mainly on culture conditions of methane oxidative strain B22 and its metabolites in methanol, ethanol. Preliminary optimization of methane bacteria B22 were obtained: 1) methane oxidative bacteria B22 best cultivate nitrogen source was potassium nitrate; 2) the optimum growth temperature was 30 ℃; 3) the optimal growth pH was 7; 4) adding vitamin complex into the culture medium. Secondly, the metabolites of strain B22 in methanol and ethanol were preliminary studied: strain B22 in aerobic environment were higher than the total growth and reproduction of anaerobic environment; formic acid and acetic acid were produced during strain B22 was in methanol and ethanol as substrates for aerobic fermentation, and acetic acid without formic acid in anaerobic fermentation produced. The research of this paper is going to be more different ways to optimize culture conditions, at the same time hope to further analysis of metabolites. 

Keywords: Methane-oxidizing bacteria, Methyl alcohol, Methane, HPLC

目录

1 前言 6

1.1 甲烷氧化菌的研究意义 6

1.2 甲烷氧化菌的菌种特性及分类 6

1.3 甲烷氧化菌的特征酶及其氧化机理 6

1.4 甲烷氧化菌的研究进展 7

1.5 本实验的设计思路 7

2 材料与方法 9

2.1 材料 9

2.1.1 菌种 9

2.1.2 实验所用培养基 9

2.1.3 实验所用试剂 9

2.1.4 实验所用仪器 10

2.2 方法 10

2.2.1 菌种的纯化培养方法 10

2.2.2 甲烷氧化菌的培养优化 10

2.2.3 甲烷氧化菌的耗氧发酵培养 11

2.2.4 甲烷氧化菌的厌氧氧发酵培养 12

2.2.5 甲烷氧化菌发酵代谢产物的液相色谱分析 12

3 结果与分析 14

3.1 不同培养条件对菌株B22生长的影响 14

3.1.1 不同N源对菌株B22生长的影响 14

3.1.2 不同pH对菌株B22生长的影响 14

3.1.3 不同温度对菌株B22生长的影响 15

3.2 不同氧环境下菌株B22生长及产物初步分析