食品与发酵工业 >

2021 , Vol. 47 >Issue 14: 51 - 56

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026394

研究报告

敲除mig1基因对马克斯克鲁维酵母利用葡萄糖和木糖的影响

  • 李俊毅 ,
  • 王大红 ,
  • 苏佳杰 ,
  • 姬翔 ,
  • 李阳
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  • 1(河南科技大学 食品与生物工程学院,河南 洛阳, 471023)
    2(河南省食品微生物工程技术研究中心,河南 洛阳, 471023)
    3(河南科技大学,微生物资源开发与利用校级重点实验室,河南 洛阳,471023)
硕士研究生(王大红副教授和李阳讲师为共同通讯作者,E-mail:wangdahong2003@163.com;liyang@haust.edu.cn)

收稿日期: 2020-12-07

  修回日期: 2021-01-20

  网络出版日期: 2021-08-20

基金资助

河南省重点攻关项目(182102210282)

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Effect of mig1 knockout on the utilization of glucose and xylose by Kluyveromyces marxianus

  • LI Junyi ,
  • WANG Dahong ,
  • SU Jiajie ,
  • JI Xiang ,
  • LI Yang
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  • 1(College of Food and Bioengineering,Henan University of Science and Technology, Luoyang 471023, China)
    2(Henan Engineering Research Center of Food Microbiology, Luoyang 471023, China)
    3(Henan University of Science and Technology, Key Laboratory of Microbial Resources Exploitation and Utilization, Luoyang 471023, China)

Received date: 2020-12-07

  Revised date: 2021-01-20

  Online published: 2021-08-20

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摘要

马克斯克鲁维酵母因其具有代谢木糖的能力而受到广泛关注。该文以具有完整木糖代谢通路的马克斯克鲁维酵母LJ0作为出发菌株,采用同源重组法敲除mig1基因,经筛选获得了敲除菌株LJ37。通过摇瓶发酵实验评价了敲除菌株的葡萄糖和木糖发酵能力,并通过RT-qPCR技术检测mig1基因的敲除对木糖代谢途径中3种关键基因表达量的影响。实验结果表明,mig1基因的敲除有效提高了菌株葡萄糖和木糖的代谢能力。以葡萄糖为底物时,敲除菌株生长量提升为野生型菌株的1.24倍,且具有利用葡萄糖进行高密度发酵的潜力。以木糖为唯一碳源时,敲除菌株的生长量提升了1.19倍;最大木糖利用速率提高到2.44 g/(L·h),为野生型的1.6倍;关键酶基因xyl1、xdh、xks1的相对表达量分别提升为野生型的2.25、2.43、2.00倍。

关键词： 马克斯克鲁维酵母; 葡萄糖效应; mig1; 基因敲除; 木糖

本文引用格式

李俊毅 , 王大红 , 苏佳杰 , 姬翔 , 李阳 . 敲除mig1基因对马克斯克鲁维酵母利用葡萄糖和木糖的影响[J]. 食品与发酵工业, 2021 , 47(14) : 51 -56 . DOI: 10.13995/j.cnki.11-1802/ts.026394

Abstract

Kluyveromyces marxianus has attracted wide attention because of its ability to metabolize xylose. In this study, the original strain was K. marxianus LJ0 with a whole xylose metabolism pathway, and the mig1 gene was knocked out by homologous recombination, and the knockout strain LJ37 was constructed. The glucose and xylose utilization ability of the knockout strain was evaluated by shaking flask fermentation experiments. RT-qPCR was used to estimate the relative expression of three key genes in xylose metabolic pathway. The results showed that the knockout strain LJ37 effectively improved the metabolic rates of glucose and xylose. The growth rate of strain LJ37 was 1.24 times that of the wild-type strain using glucose as substrate. The stain LJ37 could be applied to high-density fermentation in the future. With xylose as the sole carbon source, the growth rate of the knockout strain increased by 1.19 times, and the maximum utilization rate of xylose was enhanced to 2.44 g/(L·h), which was 1.6 times that of the wild-type strain. The RT-qPCR analysis showed that the relative expression level of genes xyl1, xdh1, and xks1 in knockout strain was 2.25, 2.43, and 2.00-fold higher than that of the wild type strain, respectively.

Key words： Kluyveromyces marxianus; glucose effect; mig1; gene knockout; xylose

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