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陈英,陈东,陆琦,芦志龙,黄日波.酿酒酵母α-半乳糖苷酶基因重组菌株的表达分析[J].广西科学,2013,20(2):143-147. [点击复制]
- CHEN Ying,CHEN Dong,LU Qi,LU Zhi-long,HUANG Ri-bo.Recombination and Expression Analysis of α-galactosidase Gene Recombinants of Saccharomyces cerevisiae[J].Guangxi Sciences,2013,20(2):143-147. [点击复制]
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| 酿酒酵母α-半乳糖苷酶基因重组菌株的表达分析 |
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陈英1,2, 陈东1,2, 陆琦1, 芦志龙1, 黄日波1
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| (1.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, 广西生物炼制重点实验室, 广西南宁 530007;2.广西大学生命科学与技术学院, 广西南宁 530004) |
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| 摘要: |
| PCR扩增里氏木霉(Trichoderma reesei)2个α-半乳糖苷酶基因agl2和agl3,将其分别与表达载体pYES2连接,电转化酿酒酵母(Saccharomyces cerevisiae)INVSc1菌株。从重组菌株提取载体进行单酶切凝胶电泳检测,证实agl2和agl3分别在重组菌株AGL2和AGL3表达。以葡萄糖和棉子糖为碳源培养菌株,2株重组菌的生长速率均显著高于原始菌株,提前8h菌数达到最大,其中以AGL3的生长速率提高较为明显,重组还使菌株在液体培养基由原来的部分絮凝转变为完全游离状。2株重组菌株均不能利用蜜二糖为碳源生长。 |
| 关键词: α-半乳糖苷酶基因 pYES2 酿酒酵母 细胞生长 细胞形态 |
| DOI： |
| 投稿时间：2013-03-10修订日期：2013-03-26 |
| 基金项目:国家973项目(2010CB736209),国家863项目(2012AA022106),国家国际合作项目(2010DFB63590,2011DFA61910),广西科学研究与技术开发计划项目(桂科合10100019-21,桂科攻1099071,桂科合1140010-15),广西自然科学基金项目(2012GXNSFAA053062),广西科学院基本科研业务费项目(10YJ25SW15),八桂学者建设工程专项经费项目资助。 |
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| Recombination and Expression Analysis of α-galactosidase Gene Recombinants of Saccharomyces cerevisiae |
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CHEN Ying1,2, CHEN Dong1,2, LU Qi1, LU Zhi-long1, HUANG Ri-bo1
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| (1.State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-Food Biorefinery, Guangxi Academy of Science, Guangxi Key Laboratory of Biorefinery, Nanning, Guangxi, 530007, China;2.Life Science and Technology College, Guangxi University, Nanning, Guangxi, 530004, China) |
| Abstract: |
| Two α-galactosidase genes agl2 and agl3,amplified respectively by PCR from the genomic DNA of Trichoderma reesei,were respectively electric transformed the S.cerevisiae strain INVSc1 using expression vector pYES2.The agl2 and agl3 were verified to be expressed respectively in the recombinant AGL2 and AGL3 by agarose electrophoresis analysis of the BamHI enzymatic digesting products of expression vectors isolated from recombinants.Cultivation in the medium with glucose or raffinose as carbon resource,the growth of both recombinants was significantly faster than that of INVSc1 strain,the maximum cell number was occurred 8 hours earlier than INVSc1,and the cell distribution in media was changed from some extent flocculation of INVSc1 to the complete free status of both recombinants.However,the result was difference with the previous investigation,neither recombinants nor INVSc1 could grow in the medium with melibiose as carbon resource.The results imply that the growth of industrial S.cerevisiae strain can be enhanced to facilitate the ethanol fermentation by transformation of α-galactosidase genes from T.reesei.Since no any relationship between α-galactosidase gene and microbial growth or cell flocculation was reported previously,the mechanism of the results should be addressed future |
| Key words: α-galactosidase gene pYES2 S.cerevisiae cell growth cell morphology |
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