基因重组构建甘蔗糖蜜酒精高产酿酒酵母菌株
发布时间：2018-11-08 10:48
【摘要】：[目的]增强工业用酿酒酵母菌株对甘蔗糖蜜中非还原糖的利用,提高糖蜜发酵乙醇产率。[方法]构建载体p YX212-agl3,分别在载体的TPI启动子和终止子5’端加上10 bp的酵母转座子重复序列,PCR扩增获得TPIP-agl3-TPIT片段,将该片段电转化至工业用野生型酿酒酵母MF1001的单倍体细胞中,复倍后经过YPR培养基初筛和YPSR培养基复筛。[结果]获得的MFA1001-3菌株分别在10锤、20锤、30锤和40锤的甘蔗糖蜜培养基中细胞的生长速率、酒精发酵浓度、对糖分的利用能力均有所增强,在40锤糖蜜中的差异最为明显,最大细胞数增加了58.7%,最大产酒率提高了4.5%,发酵结束后醪液中的总糖浓度减少了23.6%,还原糖浓度增加了15.05%。[结论]与出发菌株相比重组菌株发酵醪液中最终还原糖浓度增加而总糖浓度却减少,说明用该方法整合表达α-半乳糖苷酶的重组菌株确实有效地水解了糖蜜中的部分非还原糖同时提高了对总糖的利用率。
[Abstract]:[objective] to enhance the utilization of non-reducing sugar in sugarcane molasses by industrial Saccharomyces cerevisiae and to improve the ethanol yield of molasses fermentation. [methods] the vector p YX212-agl3, was constructed with yeast transposon repeats of 10 bp at the TPI promoter and Terminator 5'end of the vector, and TPIP-agl3-TPIT fragment was obtained by PCR amplification. The fragment was electrotransformed into the haploid cells of wild type Saccharomyces cerevisiae MF1001, and then was screened by YPR medium and YPSR medium. [results] the cell growth rate, alcohol fermentation concentration, and the utilization of sugar in the molasses medium of 10 hammers, 20 hammers, 30 hammers and 40 hammers of sugarcane molasses were increased, respectively. The difference was most obvious in 40 hammers molasses, the maximum cell number increased by 58.7, the maximum yield of wine increased by 4.5%, the total sugar concentration in the mash decreased by 23.6g and the concentration of reducing sugar increased by 15.05% after fermenting. [conclusion] compared with the original strain, the final reducing sugar concentration in the fermentation mash of the recombinant strain increased but the total sugar concentration decreased. The results showed that the recombinant strain expressing 伪 -galactosidase could effectively hydrolyze some non-reducing sugar in molasses and increase the utilization rate of total sugar.
【作者单位】： 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;广西大学生命科学与技术学院广西亚热带生物资源保护利用重点实验室;
【基金】：广西科学研究与技术开发计划项目(“木质纤维素水解技术的引进与合作研究”,No.桂科合15104001-8;“蔗渣清洁、高效转化乙醇关键技术引进与合作研究”,No.桂科合14123001-6;“蔗渣乙醇高效低成本清洁生产关键技术开发及示范”,No.桂科重14122004-3;“糖蜜乙醇高效清洁生产关键技术及示范”,No.桂科重14122004-1;“甘蔗渣产酒精新型酵母基因组研究”,No.桂科攻1517-07) 广西自然科学基金项目(“酿酒酵母工业菌株呼吸突变体生理特性及其基因变异的研究”,No.2014GXNSFAA118103) 广西科学院基本科研业务费资助项目(“基金重组构建甘蔗糖蜜酒精高效发酵菌株研究”,No.11YJ24SW10)
【分类号】：Q78;TQ223.122;TQ926
本文编号：2318234
[Abstract]:[objective] to enhance the utilization of non-reducing sugar in sugarcane molasses by industrial Saccharomyces cerevisiae and to improve the ethanol yield of molasses fermentation. [methods] the vector p YX212-agl3, was constructed with yeast transposon repeats of 10 bp at the TPI promoter and Terminator 5'end of the vector, and TPIP-agl3-TPIT fragment was obtained by PCR amplification. The fragment was electrotransformed into the haploid cells of wild type Saccharomyces cerevisiae MF1001, and then was screened by YPR medium and YPSR medium. [results] the cell growth rate, alcohol fermentation concentration, and the utilization of sugar in the molasses medium of 10 hammers, 20 hammers, 30 hammers and 40 hammers of sugarcane molasses were increased, respectively. The difference was most obvious in 40 hammers molasses, the maximum cell number increased by 58.7, the maximum yield of wine increased by 4.5%, the total sugar concentration in the mash decreased by 23.6g and the concentration of reducing sugar increased by 15.05% after fermenting. [conclusion] compared with the original strain, the final reducing sugar concentration in the fermentation mash of the recombinant strain increased but the total sugar concentration decreased. The results showed that the recombinant strain expressing 伪 -galactosidase could effectively hydrolyze some non-reducing sugar in molasses and increase the utilization rate of total sugar.
【作者单位】： 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;广西大学生命科学与技术学院广西亚热带生物资源保护利用重点实验室;
【基金】：广西科学研究与技术开发计划项目(“木质纤维素水解技术的引进与合作研究”,No.桂科合15104001-8;“蔗渣清洁、高效转化乙醇关键技术引进与合作研究”,No.桂科合14123001-6;“蔗渣乙醇高效低成本清洁生产关键技术开发及示范”,No.桂科重14122004-3;“糖蜜乙醇高效清洁生产关键技术及示范”,No.桂科重14122004-1;“甘蔗渣产酒精新型酵母基因组研究”,No.桂科攻1517-07) 广西自然科学基金项目(“酿酒酵母工业菌株呼吸突变体生理特性及其基因变异的研究”,No.2014GXNSFAA118103) 广西科学院基本科研业务费资助项目(“基金重组构建甘蔗糖蜜酒精高效发酵菌株研究”,No.11YJ24SW10)
【分类号】：Q78;TQ223.122;TQ926
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