生物工程學(xué)報(bào)  25 June 2009, 25(6):875～879

spt15突變基因?qū)︶劸平湍改咎抢玫挠绊?/p>

劉紅梅, 唐雯, 來(lái)燦鋼, 嚴(yán)明, 許琳, 歐陽(yáng)平凱

南京工業(yè)大學(xué)制藥與生命科學(xué)學(xué)院 材料化學(xué)工程國(guó)家重點(diǎn)實(shí)驗(yàn)室, 南京 210009

摘  要: 利用基因工程手段得到重組菌YPH499-3中的spt15有效突變基因, 通過(guò)表達(dá)載體 pYX212轉(zhuǎn)化入釀酒酵母原始菌株YPH499中, 重新獲得釀酒酵母重組菌株。對(duì)其性狀進(jìn)行研究, 結(jié)果表明該菌株能有效利用木糖并共發(fā)酵木糖和葡萄糖,。在30oC,、200 r/min, 發(fā)酵72 h時(shí), 50 g/L木糖的利用率為82.0%, 乙醇產(chǎn)率為28.4%; 當(dāng)木糖和葡萄糖以質(zhì)量比1:1混合發(fā)酵時(shí), 木糖和葡萄糖的利用率分別為80.4%和100%, 乙醇產(chǎn)率為31.4%; 同時(shí)發(fā)現(xiàn)木糖醇的含量極低。從而驗(yàn)證了有效突變基因spt15-10對(duì)釀酒酵母共發(fā)酵木糖和葡萄糖產(chǎn)酒精的影響,。

關(guān)鍵詞: spt15, 釀酒酵母, 木糖利用, 共發(fā)酵

Effects of mutational spt15 gene to xylose utilization of Saccharomyces cerevisiae

Hongmei Liu, Wen Tang, Cangang Lai, Ming Yan, Lin Xu, and Pingkai Ouyang

State Key Laboratory of Materials-oriented Chemical Engineering, College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China

Abstract: We used genetic methods to get a mutational spt15 gene from the recombinant strain Saccharomyces cerevisiae YPH499-3, screened by global transcription machinery engineering (gTME) approach. We transformed the gene into the original strain Saccharomyces cerevisiae YPH499 using the vector pYX212, then got a new recombinant strain. We studied the characteristic of this strain and found that it could metabolize xylose and co-ferment xylose and glucose. Under the fermentation condition of 30oC, 200 r/min, 72 h , the utilization ratio of xylose was 82.0%, with 32.4% of ethanol yield when the carbon source in the media was   50 g/L xylose, while the utilization ratio of xylose and glucose was 80.4% and 100% respectively, with the 31.4% of ethanol yield when the carbon source was 50 g/L glucose/xylose (1:1). Meanwhile, the concentration of the by-product xylitol was very low. This study demonstrates the effect which the forward mutation of spt15 gene makes to the co-fermentation of xylose and glucose to ethanol by Saccharomyces cerevisiae.

Keywords: spt15, Saccharomyces cerevisiae, xylose utilization, co-fermentation

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