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解淀粉芽孢杆菌诱变育种及其突变株在降低酱油中氨基甲酸乙酯的应用
引用本文:张梦寒,李巧玉,周朝晖,卢丽玲,堵国成,陈坚,方芳. 解淀粉芽孢杆菌诱变育种及其突变株在降低酱油中氨基甲酸乙酯的应用[J]. 微生物学报, 2017, 57(12): 1817-1826
作者姓名:张梦寒  李巧玉  周朝晖  卢丽玲  堵国成  陈坚  方芳
作者单位:江南大学生物工程学院, 江苏 无锡 214122;工业生物技术教育部重点实验室, 江苏 无锡 214122,江南大学生物工程学院, 江苏 无锡 214122;工业生物技术教育部重点实验室, 江苏 无锡 214122,广东珠江桥生物科技股份有限公司, 广东 中山 528415,广东珠江桥生物科技股份有限公司, 广东 中山 528415,江南大学生物工程学院, 江苏 无锡 214122;糖化学与生物技术教育部重点实验室, 江苏 无锡 214122,江南大学生物工程学院, 江苏 无锡 214122;粮食发酵工艺与技术国家工程实验室, 江苏 无锡 214122,江南大学生物工程学院, 江苏 无锡 214122;工业生物技术教育部重点实验室, 江苏 无锡 214122
基金项目:国家自然科学基金(31371821);广东省科技计划项目(2015B020205002)
摘    要:【目的】通过诱变育种提高解淀粉芽孢杆菌JY06利用精氨酸的能力,并将其用于降低酱油中的氨基甲酸乙酯及前体,从而提高酿造酱油的安全性。【方法】采用等离子诱变和紫外诱变两种诱变育种方法对解淀粉芽孢杆菌JY06进行突变,应用高通量筛选手段获得具有高精氨酸利用能力的突变株,验证突变株降低酱油中氨基甲酸乙酯的能力。【结果】获得了12株精氨酸利用能力提高的突变株,与出发菌株JY06相比,突变株C12和E6可使酱油中瓜氨酸含量分别降低了15.6%和14.7%,EC的含量分别降低了19.3%和13.1%。【结论】通过等离子诱变和紫外诱变进一步提高了解淀粉芽孢杆菌JY06降低酱油中EC及其前体瓜氨酸的能力,具有控制或减少酱油中生物危害物的应用潜力。

关 键 词:酱油  氨基甲酸乙酯  瓜氨酸  解淀粉芽孢杆菌  诱变育种
收稿时间:2016-12-22
修稿时间:2017-02-15

Reduction of ethyl carbamate in soy sauce by Bacillus amyloliquefaciens mutants
Menghan Zhang,Qiaoyu Li,Zhaohui Zhou,Liling Lu,Guocheng Du,Jian Chen and Fang Fang. Reduction of ethyl carbamate in soy sauce by Bacillus amyloliquefaciens mutants[J]. Acta microbiologica Sinica, 2017, 57(12): 1817-1826
Authors:Menghan Zhang  Qiaoyu Li  Zhaohui Zhou  Liling Lu  Guocheng Du  Jian Chen  Fang Fang
Affiliation:School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu Province, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu Province, China,School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu Province, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu Province, China,Guangdong Pearl River Bridge Biological Limited Corporation, Zhongshan 528415, Guangdong Province, China,Guangdong Pearl River Bridge Biological Limited Corporation, Zhongshan 528415, Guangdong Province, China,School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu Province, China;Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu Province, China,School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu Province, China;National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, Jiangsu Province, China and School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu Province, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu Province, China
Abstract:[Objective] To prepare mutants of Bacillus amyloliquefaciens JY06 stain with enhanced arginine consumption capability to reduce carcinogenic compound ethyl carbamate and its precursors in soy sauce.[Methods] Atmospheric and room temperature plasma (ARTP) and UV mutagenesis were used to mutate Bacillus amyloliquefaciens JY06, and a high-throughput screening approach was used to characterize mutants with high arginine utilization capability. The properties of mutants were studied following addition to soy sauce moromi during fermentation to initiate reduction of citrulline and ethyl carbamate.[Results] Twelve mutants with elevated arginine utilizing ability were obtained through ARTP and UV mutagenesis, of which C12 and E6 displayed the highest capacity for eliminating EC and citrulline. Compared with the JY06 parent strain, the addition of C12 or E6 to the moromi during soy sauce fermentation decreased citrulline content in soy sauce by 15.6% and 14.7%, and reduced ethyl carbamate by 19.3% and 13.1%, respectively.[Conclusion] Both ethyl carbamate and its precursor citrulline were significantly decreased in soy sauce by B. amyloliquefaciens JY06 mutants in the moromi during fermentation, demonstrating the potential of the mutants to control or eliminate toxic compounds in soy sauce and similar food products.
Keywords:soy sauce  ethyl carbamate  citrulline  Bacillus amyloliquefaciens  mutagenesis
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