Construction a Metabolic Engineering Strain to Produce 1，3-propanediol from Klebsiella pneumoniae by ldhA Gene Deletion Mutation

doi:10.13560/j.cnki.biotech.bull.1985.2015.04.017

Biotechnology Bulletin ›› 2015, Vol. 31 ›› Issue (3): 121-126.doi: 10.13560/j.cnki.biotech.bull.1985.2015.04.017

Previous Articles Next Articles

Construction a Metabolic Engineering Strain to Produce 1，3-propanediol from Klebsiella pneumoniae by ldhA Gene Deletion Mutation

Chen Lifei Li Meng Ma Chunling Yang Jianlou

(Shandong Provincial Key Laboratory of Microbial Engineering，School of Food and Bioengineering，Qilu University of Technology，Jinan 250353)

Received:2014-08-16 Online:2015-03-16 Published:2015-03-16

Abstract

Abstract: Production of 1, 3-propanediol from glycerol by Klebsiella pneumoniae is restrained by lactate formation. The increase of lactate in the broth can inhibit cell growth and reduce 1, 3-propanediol conversion rate. The lactate dehydrogenase gene(ldhA)of production lactate by Klebsiella pneumoniae was modified by λRed recombination system. One 300 bp homologous recombinant fragment：ldhAl-Cm-ldhA2 was constructed for the gene knockout. After resistance experiments, PCR determination, K. pneumonia2-1ΔldhA with ldhA gene knocked out obtained by λRed recombination. After 24 h fermentation, the maximum yield of lactate was reduced to 0.49 g/L from 10.16 g/L and the maximum threonine yield of 1, 3- propanediol was increased to 85.76 g/L from 78.83 g/L, comparing with that of the original strain K. pneumonia2-1. The conversation rate of glycerol was improved to 65.97% from 60.64%, increasing by 5.33%.

Key words: Klebsiella pneumonia, ldhA, λRed recombination, 1, 3-propanediol

Chen Lifei, Li Meng, Ma Chunling, Yang Jianlou. Construction a Metabolic Engineering Strain to Produce 1，3-propanediol from Klebsiella pneumoniae by ldhA Gene Deletion Mutation[J]. Biotechnology Bulletin, 2015, 31(3): 121-126.

References

[1] 杨菊群, 王幸宜. 1, 3-丙二醇的合成工艺进展[J]. 化学工业与工程技术, 2002, 23(2)：11-14.
[2] Biebl H, Menzel K, Zeng AP, et al. Microbial production of 1, 3-propanediol[J]. Appl Microbiol Biotechnol, 1999, 52(3)：289-297.
[3] 修志龙. 微生物发酵法生产1, 3-丙二醇的研究进展[J]. 微生物学通报, 2000, 27(4)：300-302.
[4] 张青瑞, 修志龙, 曾安平. 克雷伯氏杆菌发酵生产1, 3-丙二醇的代谢流量优化分析[J]. 化工学报, 2006, 57(6)：1403-1409.
[5] 綦文涛, 修志龙. 甘油歧化生产1, 3-丙二醇过程的代谢和基因调控机理研究进展[J]. 中国生物工程杂志, 2003, 23(2)：64-68.
[6] 刘龙飞, 周佳佳, 崔毅力, 等. 1, 3-丙二醇发酵过程中乳酸抑制的研究[J]. 生物技术, 2014, 24(1)：92-96.
[7] 莫隽颖, 陶冶, 周佳佳, 等. Red重组系统在克雷伯氏肺炎杆菌中的应用研究[J]. 化学与生物工程, 2011, 28(8)：63-66.
[8] Yamamoto S, Izumiya H, Morita M, et al. Application of λRed recombination system to Vibrio choleraegenetics：Simple methods for inactivation and modification of chromosomal genes[J]. Gene, 2009, 438(1)：57-64.
[9] 孟冬冬, 王瑞明, 马春玲. 一株1, 3-丙二醇生产菌的分离鉴定及其发酵特性[J]. 微生物学通报, 2011, 38(6)：803-808.
[10] 陈宏文. 生物法合成1, 3-丙二醇的过程工程研究[D]. 天津：天津大学, 2005.
[11] 任良栋. 克雷伯氏菌发酵生产1, 3-丙二醇工艺研究[D]. 济南：齐鲁工业大学, 2013.
[12] 魏东, 王敏, 庄文超, 等. 克雷伯氏肺炎杆菌电击转化条件的优化[J]. 中国酿造, 2012, 34(4)：18-21.
[13] Guang Y, Tian JS, Li JL. Fermentationof 1, 3-propanediol by a lactate deficient mutant of Klebsiella oxytoca under microbic conditions[J]. Appl Microbiol Biotechnol, 2007, 73(5)：1017-1024.
[14] Oh BR, Seo JW, Heo SY, et al. Optimization of culture conditions for 1, 3-propanediol production from glycerol using a mutant strain of Klebsiella pneumoniae[J]. Appl Biochem Biotechnol, 2012, 166(1)：127-137.
[15] 惠友权, 赵杰. 气相色谱法快速分析1, 3-丙二醇发酵液中醇类[J]. 应用化工, 2008, 37(8)：955-956.
[16] 牟英, 王元好, 修志龙, 等. 高效液相色谱法测定1, 3-丙二醇发酵液中的有机酸[J]. 分析化学, 2006, 34(9)：183-186.
[17] 吕吉鸿, 诸葛斌, 沈微, 等. 两步发酵过程中有机酸对产1, 3-丙二醇的影响[J]. 中国生物工程杂志, 2007, 27(5)：56-59.
[18] Oh BR, Seo JW, Heo SY, et al. Efficient production of 1, 3-propanediol from glycerol upon constitutive expression of the 1, 3-propanediol oxidoreductase gene in engineered Klebsiella pneumoniae with elimination of by-product formation[J]. Bioprocess Biosyst Eng, 2013, 36(6)：757-763.
[19] 蔡忠贞. 克雷伯氏肺炎杆菌乳酸脱氢酶基因缺失突变株的构建及其应用[D]. 乌鲁木齐：新疆大学, 2008.

Construction a Metabolic Engineering Strain to Produce 1，3-propanediol from Klebsiella pneumoniae by ldhA Gene Deletion Mutation

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 1

Recommended Articles

Metrics

Comments