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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2015, Vol. 35 Issue (9): 28-34    DOI: 10.13523/j.cb.20150905
研究报告     
比较ptaack敲除对钝齿棒杆菌产L-精氨酸生理代谢的影响
万方1, 张斌1, 陈民良2, 陈进聪2, 陈雪岚1
1 江西师范大学生命科学学院 南昌 330022;
2 南昌大学生命科学学院 南昌 330031
Comparsion of the Effects of pta and ack Deletion on Physiological Metabolism of L-arginine-producing Strain Corynebacterium crenatum
WAN Fang1, ZHANG Bin1, CHEN Min-liang2, CHEN Jin-cong2, CHEN Xue-lan1
1 College of Life Science, Jiangxi Normal University, Nanchang 330022, China;
2 College of Life Science, Nanchang University, Nanchang 330031, China
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摘要:

旁支代谢途径的截断有利于目的氨基酸合成途径的集流.基于基因组尺度代谢网络模型的预测,以钝齿棒杆菌(Corynebacterium crenatum)MT-M4为出发菌株,通过无痕敲除技术分别敲除了编码磷酸乙酰基转移酶的pta基因及编码乙酸激酶的ack基因,阻断了乙酸的合成.摇瓶发酵结果表明,pta缺失菌株精氨酸产量较出发菌株提高了25.60%,达15.46g/L.葡萄糖转化率提高了29.41%;ack缺失菌株精氨酸产量达13.82g/L,较出发菌株提高了12.81%,葡萄糖转化率提高了26.02%.同时,ptaack敲除菌株的细胞生长较出发菌株均分别提高了9.19%及7.71%.因此,pta、ack的敲除不仅有利于精氨酸的合成,而且对菌体生长具有促进作用;但pta的敲除更有利于精氨酸的积累.

关键词: 钝齿棒杆菌精氨酸无痕敲除技术基因组尺度代谢网络模型生理代谢    
Abstract:

The disruption of competitive metabolic pathways is conducive to the carbon flux converted into the purpose amino acid biosynthetic pathway. In the present study, based on the prediction of genome-scale metabolic network model, markless knock-out technology to construct separately recombinant strains C. crenatum MT-M4 △pta and C. crenatum MT-M4 △ack which led to interdict the acetic acid biosynthetic pathway were employed. As shake-flask fermentation results shown, L-arginine production of C. crenatum MT-M4 △pta was significantly increased by 25.60% higher than that of original strain, reached to 15.46g/L with glucose transformation rate increased by 29.41%. L-arginine production of C. crenatum MT-M4 △ack was 13.82g/L which was 12.81% higher than that of original strain, with glucose transformation rate increased by 26.02%. In addition, the growth of C. crenatum MT-M4 △pta and C. crenatum MT-M4 △ack was increased by 9.19% and 7.71%, respectively. Thus, pta and ack deletion was not only beneficial to the improvement of L-arginine production, but also conducive to the cell growth. However, pta deletion compared with ack deletion was more conducive to L-arginine accumulation.

Key words: C. crenatum    L-arginine    Markerless knock-out technology    Genome-scale metabolic network model    Physiological metabolism
收稿日期: 2015-05-18 出版日期: 2015-09-25
ZTFLH:  Q812  
基金资助:

国家自然科学基金资助项目(31360219,30960012)

通讯作者: 陈雪岚     E-mail: xuelanchen162@163.com
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引用本文:

万方, 张斌, 陈民良, 陈进聪, 陈雪岚. 比较ptaack敲除对钝齿棒杆菌产L-精氨酸生理代谢的影响[J]. 中国生物工程杂志, 2015, 35(9): 28-34.

WAN Fang, ZHANG Bin, CHEN Min-liang, CHEN Jin-cong, CHEN Xue-lan. Comparsion of the Effects of pta and ack Deletion on Physiological Metabolism of L-arginine-producing Strain Corynebacterium crenatum. China Biotechnology, 2015, 35(9): 28-34.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150905        https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I9/28


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