【摘要】：[目的]增強工業(yè)用釀酒酵母菌株對甘蔗糖蜜中非還原糖的利用,提高糖蜜發(fā)酵乙醇產(chǎn)率。[方法]構(gòu)建載體p YX212-agl3,分別在載體的TPI啟動子和終止子5’端加上10 bp的酵母轉(zhuǎn)座子重復序列,PCR擴增獲得TPIP-agl3-TPIT片段,將該片段電轉(zhuǎn)化至工業(yè)用野生型釀酒酵母MF1001的單倍體細胞中,復倍后經(jīng)過YPR培養(yǎng)基初篩和YPSR培養(yǎng)基復篩。[結(jié)果]獲得的MFA1001-3菌株分別在10錘、20錘、30錘和40錘的甘蔗糖蜜培養(yǎng)基中細胞的生長速率、酒精發(fā)酵濃度、對糖分的利用能力均有所增強,在40錘糖蜜中的差異最為明顯,最大細胞數(shù)增加了58.7%,最大產(chǎn)酒率提高了4.5%,發(fā)酵結(jié)束后醪液中的總糖濃度減少了23.6%,還原糖濃度增加了15.05%。[結(jié)論]與出發(fā)菌株相比重組菌株發(fā)酵醪液中最終還原糖濃度增加而總糖濃度卻減少,說明用該方法整合表達α-半乳糖苷酶的重組菌株確實有效地水解了糖蜜中的部分非還原糖同時提高了對總糖的利用率。
[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 偽 -galactosidase could effectively hydrolyze some non-reducing sugar in molasses and increase the utilization rate of total sugar.
【作者單位】： 廣西科學院非糧生物質(zhì)酶解國家重點實驗室國家非糧生物質(zhì)能源工程技術研究中心廣西生物質(zhì)產(chǎn)業(yè)化工程院廣西生物煉制重點實驗室;廣西大學生命科學與技術學院廣西亞熱帶生物資源保護利用重點實驗室;
【基金】：廣西科學研究與技術開發(fā)計劃項目(“木質(zhì)纖維素水解技術的引進與合作研究”,No.桂科合15104001-8;“蔗渣清潔、高效轉(zhuǎn)化乙醇關鍵技術引進與合作研究”,No.桂科合14123001-6;“蔗渣乙醇高效低成本清潔生產(chǎn)關鍵技術開發(fā)及示范”,No.桂科重14122004-3;“糖蜜乙醇高效清潔生產(chǎn)關鍵技術及示范”,No.桂科重14122004-1;“甘蔗渣產(chǎn)酒精新型酵母基因組研究”,No.桂科攻1517-07) 廣西自然科學基金項目(“釀酒酵母工業(yè)菌株呼吸突變體生理特性及其基因變異的研究”,No.2014GXNSFAA118103) 廣西科學院基本科研業(yè)務費資助項目(“基金重組構(gòu)建甘蔗糖蜜酒精高效發(fā)酵菌株研究”,No.11YJ24SW10)
【分類號】：Q78;TQ223.122;TQ926

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