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当前位置: 首页 > 文章 > 腐乳中优势细菌群落研究及丁酸梭菌的分离鉴定 河北农业大学学报 2019 (5) 79-84
Position: Home > Articles > 腐乳中优势细菌群落研究及丁酸梭菌的分离鉴定 Journal of Agricultural University of Hebei 2019 (5) 79-84

腐乳中优势细菌群落研究及丁酸梭菌的分离鉴定

作  者：
马艳莉;梁静静;丁玉峰;李素萍;席晓丽;王颉;郭书贤
单  位：
南阳理工学院/河南省工业微生物资源与发酵技术重点实验室;河北农业大学食品科技学院
关键词：
腐乳;PCR-DGGE;丁酸;丁酸梭菌
摘  要：
本研究借助聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术分析青方、红方、白方和低盐红腐乳的优势细菌群落,使用亨盖特厌氧滚管法对青方腐乳中丁酸产生菌进行分离,并进行分子生物学鉴定和生理生化特性研究.DGGE结果表明:青方腐乳的优势细菌群落多于其他类型腐乳,主要有枯草芽孢杆菌、乳酸菌、产气肠杆菌、丁酸梭菌等几大类.为了进一步利用青方腐乳中的丁酸梭菌,增强青方腐乳微生态健康作用,从青方腐乳中分离到12株单菌落,通过高效液相色谱对其培养液中丁酸含量进行测定.结果表明,12株菌中有2株菌(BP01和BP02)代谢产物含丁酸,BP01菌株和BP02菌株发酵48 h培养液中丁酸含量分别为6.48和3.27 mmol/L.通过分子生物学技术鉴定BP01菌株为丁酸梭菌,BP02为双酶梭菌.考虑到BP01菌株产丁酸能力较强且为广泛认可的益生菌,后续试验选择BP01菌株为研究对象,观察其菌株菌落形态并进行生理生化鉴定,进一步验证BP01菌为丁酸梭菌(Clostridium butyricum).
作  者：
MA Yanli;LIANG Jingjing;DING Yufeng;LI Suping;XI Xiaoli;WANG Jie;GUO Shuxian;College of Food Science and Technology, Hebei Agricultural University;Nanyang Institute of Technology/Henan Key Laboratory of Industrial Microbial Resources and Fermentation Technology;
关键词：
sufu;;PCR-DGGE;;butyric acid;;Clostridium butyricum
摘  要：
In this study, polymerase chain reaction denaturing gradient gel electrophoresis(PCR-DGGE) was used to analyze the dominant bacterial communities in grey, red, white and low-salt red sufu. Hungate anaerobic separation method was used to isolate butyric acid producing bacteria from grey sufu, and molecular biological identification and physiological and biochemical characteristics were studied. DGGE results showed that the dominant bacterial community of grey sufu was more than that of other types of sufu, including Bacillus subtilis, lactic acid bacteria, Enterobacter aerogenes and Clostridium butyricum. In order to further utilize Clostridium butyricum in grey sufu and enhance the microecological health of grey sufu, 12 single colonies were isolated from grey sufu. The butyric acid content in culture medium was determined by high performance liquid chromatography. The results showed that two of the 12 strains produced butyric acid in their metabolites, and the butyric acid contents produced by BP01 and BP02 strains were 6.48 mmol/L and 3.27 mmol/L, respectively. The strain BP01 was identified as Clostridium butyricum and BP02 as Clostridium dizymase by molecular biological techniques. Considering that BP01 strain has strong butyric acid production ability and is widely recognized as probiotic bacteria, BP01 strain was selected as the research object in the follow-up experiment. The colonial morphology of BP01 strain was observed and its physiological and biochemical identification was carried out, which further verified that BP01 strain was Clostridium butyricum.

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