外源添加乙偶姻对酿酒酵母产2,3-丁二醇及其菌株的影响

doi:10.11924/j.issn.1000-6850.casb20200300177

中国农学通报 ›› 2021, Vol. 37 ›› Issue (2): 20-27.doi: 10.11924/j.issn.1000-6850.casb20200300177

所属专题：生物技术

外源添加乙偶姻对酿酒酵母产2,3-丁二醇及其菌株的影响

张弛¹^,²(), 吕雨泽¹^,², 邓利廷¹^,², 孙健¹^,², 葛菁萍¹^,²()

¹黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨 150500
²微生物黑龙江省高校重点实验室/黑龙江大学生命科学学院,哈尔滨 150080

收稿日期:2020-03-06 修回日期:2020-05-03 出版日期:2021-01-15 发布日期:2021-01-14
通讯作者: 葛菁萍
作者简介:张弛,男,1996年出生,黑龙江鹤岗人,硕士研究生,研究方向：微生物资源挖掘与利用。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号,Tel：0451-86609016,E-mail：zhangchi0102@126.com。
基金资助:
国家自然科学基金面上项目“乙酸溢流代谢与群体感应的协同效应对副干酪乳杆菌HD1.7种群稳定性影响机制的探究”(32071519);黑龙江省自然科学基金重点项目“乙酸代谢与副干酪乳杆菌群体感应互作探究及对细菌素产生的影响”(ZD2020C008);黑龙江省现代农业产业体系协同创新推广体系麻类（工业）项目

Effects on 2,3-butanediol Produced by Saccharomyces cerevisiae and Its Strains: Acetoin Addition

Zhang Chi¹^,²(), Lv Yuze¹^,², Deng Liting¹^,², Sun Jian¹^,², Ge Jingping¹^,²()

¹Agricultural Microorganisms Technology Education Engineering Research Center, Harbin 150500
²Key Laboratory of Microbiology of Heilongjiang Province, Life Science College, Heilongjiang University, Harbin 150080

Received:2020-03-06 Revised:2020-05-03 Online:2021-01-15 Published:2021-01-14
Contact: Ge Jingping

摘要/Abstract

摘要：

通过对菌株发酵特性的研究,旨在获得一株性能良好的2,3-丁二醇(2,3-BD)生产菌株。本研究以酿酒酵母(Saccharomyces cerevisiae)W5/W5-$\Delta$pdc1/W5-$\Delta$pdc5/W141四株菌株为试材,利用高效液相色谱法(HPLC)对乙偶姻产量和BDH酶活进行检测,从而挑选出最适合作为产2,3-BD出发菌株,并外源添加不同浓度的乙偶姻,测定乙偶姻、葡萄糖和2,3-BD产量随发酵时间的变化情况。结果表明：S. cerevisiae W5与S. cerevisiae W141最适合作为产2,3-BD出发菌株,且S. cerevisiae W5最适添加量为12 g/L乙偶姻,且72 h时,2,3-BD的产量最大达到2.49 ± 0.02 g/L;S. cerevisiae W141最适添加量为8 g/L乙偶姻,且72 h时,2,3-BD的产量最大达到1.56 ± 0.04 g/L,外源添加乙偶姻后菌株生长状况良好,并且二者产生的乙偶姻在发酵结束前消耗完全,同时在24 h左右,二者产生的葡萄糖也均基本耗尽。这为研究和利用2,3-BD等可再生资源提供了思路,也为利用S. cerevisiae生产2,3-BD奠定了理论基础。

关键词: 2,3-丁二醇, 乙偶姻, 酿酒酵母, 菌株生长, HPLC

Abstract:

By studying the fermentation characteristics of strain, we aim at obtaining a good 2,3-butanediol (2,3-BD) producing strain. Four strains of Saccharomyces cerevisiae W5/W5-$\Delta$pdc1/W5-$\Delta$pdc5/W141 were used as materials in this study, the yield of acetoin and BDH enzyme activity were detected with HPLC, in order to select the most optimum starting strain for producing 2,3-BD, and the starting strain producing 2,3-BD was added with different concentrations of acetoin, and the changes of acetoin, glucose and 2,3-BD production with fermentation time were determined. The result showed that S. cerevisiae W5/W141 were selected as the optimum strain, the optimum acetoin concentration of S. cerevisiae W5 was 12 g/L, and the concentration of 2,3-BD reached the maximum at 72 h, and it was 2.49 ± 0.02 g/L. The optimum acetoin concentration of S. cerevisiae W141 was 8 g/L, and the concentration of 2,3-BD reached the maximum at 72 h, and it was 1.56 ± 0.04 g/L, the strain grew well after adding acetoin, and the acetoin produced by S. cerevisiae W5/W141 was completely consumed before the end of fermentation, and the glucose produced by S. cerevisiae W5/W141 was almost exhausted at 24 h. This provides an idea for the research and utilization of renewable resources such as 2,3-BD, and also lays a theoretical foundation for S. cerevisiae produces 2,3-BD.

Key words: 2,3-butanediol, acetoin, Saccharomyces cerevisiae, strain growth, HPLC

中图分类号:

S182

张弛, 吕雨泽, 邓利廷, 孙健, 葛菁萍. 外源添加乙偶姻对酿酒酵母产2,3-丁二醇及其菌株的影响[J]. 中国农学通报, 2021, 37(2): 20-27.

Zhang Chi, Lv Yuze, Deng Liting, Sun Jian, Ge Jingping. Effects on 2,3-butanediol Produced by Saccharomyces cerevisiae and Its Strains: Acetoin Addition[J]. Chinese Agricultural Science Bulletin, 2021, 37(2): 20-27.

图/表 8

图1. 乙偶姻产量和BDH酶活检测结果

图2. S. cerevisiae W5/W141的葡萄糖消耗量随发酵时间变化的关系曲线

图3. S. cerevisiae W5/W141的乙偶姻剩余量和2,3-丁二醇产量随发酵时间变化的关系曲线

图4. 不同时间段流式细胞术检测结果 A1~A6和B1~B6分别代表S. cerevisiae W141/W141-8 g/L从12 h发酵到72 h的6个取样点

图5. 流式细胞术检测结果

图6. 不同时间段流式细胞术检测结果 A1-A6和B1-B6分别代表S. cerevisiae W5/W5-12 g/L从12 h发酵到72 h的6个取样时间点

图7. 流式细胞术检测结果

图8. 电镜观察结果 A1A2：S. cerevisiae W141/W141-8 g/L;B1B2：S. cerevisiae W5/W5-12 g/L

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外源添加乙偶姻对酿酒酵母产2,3-丁二醇及其菌株的影响

Effects on 2,3-butanediol Produced by Saccharomyces cerevisiae and Its Strains: Acetoin Addition

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