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黄艳燕,郭铃,陈东,龙思宇,李检秀,陆琦,孙靓,黄日波.酿酒酵母YBR019C基因缺失突变的分析[J].广西科学,2014,21(2):108-114. [点击复制]
- HUANG Yan-yan,GUO Ling,CHEN Dong,LONG Si-yu,LI Jian-xiu,LU Qi,SUN Liang,HUANG Ri-bo.Reseach on Saccharomyces cerevisiae Mutant Deficient in YBR019C[J].Guangxi Sciences,2014,21(2):108-114. [点击复制]
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| 酿酒酵母YBR019C基因缺失突变的分析 |
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黄艳燕, 郭铃, 陈东, 龙思宇, 李检秀, 陆琦, 孙靓, 黄日波
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| (广西科学院, 非粮生物质酶解国家重点实验室, 国家非粮生物质能源工程技术研究中心, 广西生物质产业化工程院, 广西生物炼制重点实验室, 广西南宁 530007) |
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| 摘要: |
| [目的]研究目的基因YBR019C缺失对酿酒酵母(Saccharomyces cerevisiae)菌株糖代谢和乙醇发酵的影响。[方法]以酿酒酵母野生菌NF1002为出发菌株,选择2号染色体上的基因YBR019C为目的基因,以质粒pUG6和pUG66为模板进行PCR,构建带有Cre/loxP系统的酿酒酵母YBR019C基因敲除组件,并转化酿酒酵母NF1002,利用筛选标记Kanr和Bler与YBR019C基因进行同源重组,筛选YBR019C双倍体缺陷型菌株。利用蔗糖和甘蔗糖蜜为碳源,对突变菌进行发酵特性的研究。[结果]成功获得YBR019C双倍体缺陷型菌株NF-ybr。碳源同化实验表明,突变株和野生菌均能利用葡萄糖和蔗糖,不能利用乳糖和木糖;但相比野生菌,突变株利用棉子糖和麦芽糖的能力有所下降,而且完全不能利用半乳糖。蔗糖发酵实验表明:突变株NF-ybr与野生菌株相比,在发酵终点乙醇浓度提高10.7%,发酵周期有所延长。按目前甘蔗糖蜜乙醇生产的发酵工艺,突变株在30℃发酵72h的醪液乙醇含量为12.52%,低于野生菌的13.89%。[结论]YBR019C基因的缺失影响了菌株对糖份的利用,导致乙醇发酵能力不及野生菌。本研究为菌株高效快捷的基因改造提供了参考。 |
| 关键词: 酿酒酵母 YBR019C 基因敲除 甘蔗糖蜜 乙醇 |
| DOI： |
| 投稿时间：2013-11-17修订日期：2013-12-17 |
| 基金项目:广西科学院基本科研业务项目(基金编号:11YJ24SW06);八桂学者建设工程专项经费项目资助。 |
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| Reseach on Saccharomyces cerevisiae Mutant Deficient in YBR019C |
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HUANG Yan-yan, GUO Ling, CHEN Dong, LONG Si-yu, LI Jian-xiu, LU Qi, SUN Liang, HUANG Ri-bo
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| (Guangxi Academy of Sciences, State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-Food Biorefinery, Guangxi Biomass Industrialization Engineering Institute, Guangxi Key Laboratory of Biorefinery, Nanning, Guangxi, 530007, China) |
| Abstract: |
| [Objective] Our study focuses on the saccharometabolism and ethanol fermentation of Saccharomyces cerevisiae strain NF1002.[Methods] YBR019C on Chromosome Ⅱ was chosen to be modified by PCR with pUG6 and pUG66 plasmids as templates.After homologous recombination of Cre/loxP mediated marker and YBR019C, YBR019C deficient mutant of Saccharomyces cerevisiae strain NF1002 was constructed.[Results] Glucose and sucrose can be utilized for metabolism at both mutant and wild strains except lactose and xylose.However, only part of raffinose and maltose and few galactose can be utilized at the mutant strain.Results of sucrose fermentation at mutant strain NF-ybr displayed that ethanol content reached 13.68%(V/V)at the end of fermentation, which was 10.7% higher than that for the wild strain.Furthermore, according to the industrial processes for ethanol fermentation of sugarcan molasses, ethanol content was 12.015%(V/V)at 30℃ for 72h, lower than that of the wild strain.[Conclusion] Saccharometabolism of Saccharomyces cerevisiae strain NF1002 was affected by deletion of gene YBR019C, showing less ethanol production.Our research provided advice on modification of Saccharomyces cerevisiae strains on an efficient way. |
| Key words: Saccharomyces cerevisiae YBR019C gene deletion sugarcan molasses ethanol |
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