<em>ack</em>基因敲除对<em>Thermoanaerobacterium calidifontis</em> Rx1发酵代谢的影响

doi:10.13523/j.cb.20150706

中国生物工程杂志

2015, Vol. 35

Issue (7): 37-44 DOI: 10.13523/j.cb.20150706

研究报告

ack基因敲除对Thermoanaerobacterium calidifontis Rx1发酵代谢的影响

申冬玲¹, 尚淑梅², 李卫娜¹, 严金平¹, 伊日布斯¹

1. 昆明理工大学生命科学与技术学院昆明 650500;
2. 长江师范学院生命科学与技术学院重庆 408100

Characterization of the Disrupted ack Genes on Fermentation by Thermoanaerobacterium calidifontis Rx1

SHEN Dong-ling¹, SHANG Shu-mei², LI Wei-na¹, YAN Jin-ping¹, HANGAN Ir-bis¹

1. Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China;
2. Life Science and Technology Institute, Yangtze Normal University, Chongqing 408100, China

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摘要：

为提高嗜热厌氧菌T. calidifontis Rx1的乙醇产率,通过同源重组的方式敲除Rx1乙酸生成途径的乙酸激酶(ack)基因,得到了代谢工程菌Δack突变株。分别以葡萄糖、纤维二糖、木糖和玉米芯酸水解液为底物,研究突变菌株的底物利用、细胞生长和发酵产物的变化情况。结果表明,与野生菌株相比,突变菌株的干重都有所降低,但是乳酸或乙醇的得率显著提高;当以纤维二糖为底物时,突变菌株的乙醇产量达3.60g/L,得率为0.55g/g,远高于突变菌株对其它底物的产量;以玉米芯水解液为底物时,突变菌株的乳酸产量高于野生菌株,而且野生菌乳酸、乙醇的产量都高于以木糖为底物时的产量。

关键词： 高温厌氧菌; 基因敲除; 乙醇发酵

Abstract:

To improve the yield of ethanol by Thermoanaerobacterium calidifontis Rx1, constructed a engineered mutant Δack. First a recombinant plasmid containing mutation cassettes of pta::ack, was rebuild, and the vector was transformed to cell to disrupt the target genes on the chromosomal via the homologous recombination. Then glucose fermentation, cellobiose fermentation, xylose fermentation, acid hydrolyzate of corncob of Δack mutant and the wild strain were performed respectively to produce ethanol and lactate. The results indicate that the acetate of Δack mutant is much lower as compared with the wild. Dray cell weight of the mutant is always lower than that of the wild under four conditions. However, the yield of ethanol or lactate is more than the wild. When Δack mutant used cellobiose to produce ethanol, the yield is 3.60g/L higher than another three substrates. At the same time, it could be exist approximative 40mmol/L acetate in the hydrolysate, so the output of lactate and ethanol of the wild are more than that with xylose fermentation.

Key words: Thermophilic anaerobic bacteria Gene knockout Ethanol fermentation

收稿日期: 2015-02-26 出版日期: 2015-07-25

ZTFLH:

Q789

基金资助:

国家自然科学基金资助项目(21366015)

通讯作者: 伊日布斯 E-mail: irbisc@gmail.com

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引用本文:

申冬玲, 尚淑梅, 李卫娜, 严金平, 伊日布斯. ack基因敲除对Thermoanaerobacterium calidifontis Rx1发酵代谢的影响[J]. 中国生物工程杂志, 2015, 35(7): 37-44.

SHEN Dong-ling, SHANG Shu-mei, LI Wei-na, YAN Jin-ping, HANGAN Ir-bis. Characterization of the Disrupted ack Genes on Fermentation by Thermoanaerobacterium calidifontis Rx1. China Biotechnology, 2015, 35(7): 37-44.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150706 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I7/37

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