一株耐受高浓度葡萄糖且产乙偶姻菌株的筛选及产物鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2020-0119

生物技术通报 ›› 2020, Vol. 36 ›› Issue (8): 53-60.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0119

一株耐受高浓度葡萄糖且产乙偶姻菌株的筛选及产物鉴定

霍明月^1,2, 张鸽², 苏玉龙³, 李兴², 张海波², 峥嵘¹, 刘好宝^4,5

1.内蒙古师范大学生命科学与技术学院,呼和浩特 010022;
2.中国科学院青岛生物能源与过程研究所生物基材料重点实验室,青岛 266101;
3.云南省烟草公司玉溪市公司元江县分公司,玉溪 6531005;
4.中国农业科学院烟草研究所农业部烟草生物学与加工重点实验室,青岛 266101;
5.中国烟草总公司海南省公司海口雪茄研究所,海口 571100

收稿日期:2020-02-10 出版日期:2020-08-26 发布日期:2020-08-27
作者简介:霍明月,女,硕士研究生,研究方向：生物化学与分子生物学;E-mail：1563033250@qq.com
基金资助:
中国烟草总公司海南省公司科技重点项目（201746000024052）,中国烟草总公司海南省公司科技重点项目（201746000024056）

Isolation of an Acetoin-producing Strain Resistant to High Concentration Glucose and Identification of Metabolites

HUO Ming-yue^1,2, ZHANG Ge², SU Yu-long³, LI Xing², ZHANG Hai-bo², ZHENG Rong¹, LIU Hao-bao^4,5

1. Inner Mongolia Normal University,College of Life Science and Technology,Huhhot 010022;
2. CAS Key Laboratory of Biobased Materials,Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Science,Qingdao 266101;
3. Yuxi City Company Yuanjiang County Branch of Yunnan Tobacco Company,Yuxi 653100;
4. Key Laboratory of Tobacco Biology and Processing,Tobacco Research Institute of Chinese Academy of Agricultural Sciences,Qingdao 266101;
5. Cigar Research Institute of Chinese Tobacco Corporation Hainan Province,Haikou 571100

Received:2020-02-10 Published:2020-08-26 Online:2020-08-27

摘要/Abstract

摘要： 为获得一株耐受高浓度葡萄糖且产乙偶姻的菌株,利用高糖培养基以及乙偶姻显色液从烟田土壤中筛选到一株全新菌株;将筛选出的菌株利用全基因组测序技术和16S rDNA 序列比对分析对菌株进行鉴定,并利用不同浓度葡萄糖的培养基对菌株进行耐受实验;最后,利用GC-MS（气相-质谱）将菌株的发酵产物定性分析,并对菌株进行72 h的初步发酵。结果显示,筛选获得菌株为金黄色葡萄球菌（Staphylococcus aureus）,命名为PX03。葡萄糖耐受实验中,菌株PX03在含有500 g/L葡萄糖的培养基中依然能生长,OD₆₀₀可达65;300 g/L葡萄糖浓度下生长最好,OD₆₀₀可达100。GC-MS定性结果分析发现,该菌株可产大量的乙偶姻（22.51%）和2,3-丁二醇（21.29%）,同时可产乙酸（13.97%）、3,4-二羟基-3,4-二甲基己烷-2,5-二酮（8.52%）、吡喃酮（5.24%）和3-甲基-丁酸（4.47%）等一些香气成分。对菌株的初步发酵结果显示,该菌株乙偶姻产量可达41 g/L。

关键词: 微生物发酵, 葡萄球菌, 乙偶姻, 葡萄糖耐受

Abstract: In order to obtain an acetoin-producing strain that is resistant to high concentrations of glucose,a new strain was screened from tobacco field soil using high-sugar medium and acetoin coloring solution;then the screened strain was sequenced and identified by whole genome technology and 16S rDNA sequence alignment analysis,and tolerance of the strain was tested at different concentrations of glucose in the medium. Finally,the GC-MS（gas chromatography-mass spectrometry）was used to qualitatively analyze the fermentation products of the strain and the capacity of acetoin-producing was analyzed by preliminary fermentation for 72 h. The results showed that the strain obtained was Staphylococcus aureus and named as PX03. In the glucose tolerance test,the strain PX03 still grew in a medium containing 500 g/L glucose,with an OD₆₀₀ of 65;grew the best at a concentration of 300 g/L glucose,with an OD₆₀₀ of 100. GC-MS qualitative analysis revealed that the strain produced a large amount of aroma components,including acetoin（22.51%）,2,3-butanediol（21.29%）,acetic acid（13.97%）,3,4-dihydroxy-3,4-dimethyl-2,5-hexanedione（8.52%）,pyranone（5.24%）and 3-methyl-bbutanoic acid（4.47%）. The preliminary fermentation results demonstrated that the yield of acetoin of this strain reached 41 g/L.

Key words: microbial fermentation, Staphylococcus, acetoin, high glucose tolerance

霍明月, 张鸽, 苏玉龙, 李兴, 张海波, 峥嵘, 刘好宝. 一株耐受高浓度葡萄糖且产乙偶姻菌株的筛选及产物鉴定[J]. 生物技术通报, 2020, 36(8): 53-60.

HUO Ming-yue, ZHANG Ge, SU Yu-long, LI Xing, ZHANG Hai-bo, ZHENG Rong, LIU Hao-bao. Isolation of an Acetoin-producing Strain Resistant to High Concentration Glucose and Identification of Metabolites[J]. Biotechnology Bulletin, 2020, 36(8): 53-60.

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一株耐受高浓度葡萄糖且产乙偶姻菌株的筛选及产物鉴定

Isolation of an Acetoin-producing Strain Resistant to High Concentration Glucose and Identification of Metabolites

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