潘柯莉,茅以诚,岳圣杰,姚瑞莲,胡洪波

• 1:微生物代谢国家重点实验室上海交通大学生命科学技术学院
• 2:复旦大学药学院

摘要(Abstract)：

木质纤维素是地球上最丰富的碳源,可广泛应用于生物燃料和生物化工。但是由于分解代谢阻遏,不能完全利用木质纤维素降解产物中获得的多种糖类,严重制约了木质纤维素的商业应用。研究主要构建了一株可以同时消耗葡萄糖、果糖、木糖的菌株E.coli BW25113△pgi,并探讨了其分别在好氧和厌氧条件下以葡萄糖、果糖、木糖为混合碳源时的碳源利用情况。敲除pgi后,葡萄糖、果糖、木糖三种糖被同时消耗。研究通过发酵数据、基因转录水平数据和代谢流分析数据阐述了敲除pgi解除分解代谢阻遏的机理。pgi基因的敲除导致pts G转录水平下降,增加了c AMP-Crp的含量,从而解除分解代谢阻遏,实现了混合碳源的共利用,为木质纤维素的开发利用提供了新的思路。

关键词(KeyWords)： 大肠杆菌;pgi敲除;碳源共利用;代谢调控

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学青年基金(31400086,81402956);; 中国博士后科学基金(2016M601584);; 上海市科委晨光计划(14CG03)

作者(Author): 潘柯莉,茅以诚,岳圣杰,姚瑞莲,胡洪波

参考文献(References)：

• [1]Guerriero G,Hausman J F,Strauss J,et al.Lignocellulosic biomass:biosynthesis,degradation,and industrial utilization[J].Engineering in Life Sciences,2016,16(1):1-16.
• [2]Luo Y,Zhang T,Wu H.The transport and mediation mechanisms of the common sugars in Escherichia coli[J].Biotechnology Advances,2014,32(5):905-919.
• [3]Liang L,Liu R,Li F,et al.Repetitive succinic acid production from lignocellulose hydrolysates by enhancement of ATP supply in metabolically engineered Escherichia coli[J].Bioresource Technology,2013,143(6):405-412.
• [4]Yao R,Hirose Y,Sarkar D,et al.Catabolic regulation analysis of Escherichia coli and its crp,mlc,mgs A,pgi and pts Gmutants[J].Microbial Cell Factories,2011,10:67.
• [5]Xia T,Eiteman M A,Altman E.Simultaneous utilization of glucose,xylose and arabinose in the presence of acetate by a consortium of Escherichia coli strains[J].Microbial Cell Factories,2012,11:77.
• [6]Deutscher J,Francke C,Postma P W.How phosphotransferase system-related protein phosphorylation regulates carbohydrate metabolism in bacteria[J].Microbiology&Molecular Biology Reviews,2006,70(4):939-1031.
• [7]Mannan A A,Toya Y,Shimizu K,et al.Integrating kinetic model of E.coli with genome scale metabolic fluxes overcomes its open system problem and reveals bistability in central metabolism[J].Plo S One,2015,10(10):e0139507.
• [8]Kim S M,Choi B Y,Ryu Y S,et al.Simultaneous utilization of glucose and xylose via novel mechanisms in engineered Escherichia coli[J].Metabolic Engineering,2015,30:141-148.
• [9]Shiue E,Brockman I M,Prather K L.Improving product yields on D-glucose in Escherichia coli via knockout of pgi andzwf and feeding of supplemental carbon sources[J].Biotechnology and Bioengineering,2015,112(3):579-587.
• [10]Fong S S,Nanchen A,Palsson B O,et al.Latent pathway activation and increased pathway capacity enable Escherichia coli adaptation to loss of key metabolic enzymes[J].Journal of Biological Chemistry,2006,281(12):8024-8033.
• [11]Lee H C,Kim J S,Jang W,et al.High NADPH/NADP+ratio improves thymidine production by a metabolically engineered Escherichia coli strain[J].Journal of Biotechnology,2014,149(1-2):24-32.
• [12]Sundara Sekar B,Seol E,Park S.Co-production of hydrogen and ethanol from glucose in Escherichia coli by activation of pentose-phosphate pathway through deletion of phosphoglucose isomerase(pgi)and overexpression of glucose-6-phosphate dehydrogenase(zwf)and 6-phosphogluconate dehydrogenase(gnd)[J].Biotechnology for Biofuels,2017,10:85.
• [13]XIONG De-wang(熊得王),LIU Qing(刘晴),YAO Rui-lian(姚瑞莲),et al.Effect of crp gene mutation on the utilization mechanism of Escherichia coli for a mixture of multiple carbon sources(Crp基因突变对大肠杆菌利用混合碳源的影响)[J].Journal of Chemical Engineering of Chinese Universities(高校化学工程学报),2015,29(6):1390-1398.
• [14]Canonaco F,Hess T A,Heri S,et al.Metabolic flux response to phosphoglucose isomerase knock-out in Escherichia coli and impact of overexpression of the soluble transhydrogenase Udh A[J].FEMS Microbiology Letters,2001,204(2):247-252.
• [15]Reiner A M.Xylitol and D-arabitol toxicities due to derepressed fructose,galactitol,and sorbitol phosphotransferases of Escherichia coli[J].Journal of Bacteriology,1977,132(1):166-173.
• [16]Usui Y,Hirasawa T,Furusawa C,et al.Investigating the effects of perturbations topgi and eno gene expression on central carbon metabolism in Escherichia coli using 13C metabolic flux analysis[J].Microbial Cell Factories,2012,11:87.
• [17]Park Y H,Lee B R,Seok Y J,et al.In vitro reconstitution of catabolite repression in Escherichia coli[J].Journal of Biological Chemistry,2006,281(10):6448-6454.
• [18]Saier M H Jr,Ramseier T M,Reizer J.Regulation of carbon utilization[C]//Escherichia coli and Salmonella:cellular and molecular biology,2nd ed.Washington DC:ASM Press,1996:1325-1343.
• [19]Saier M H Jr,Ramseier T M.The catabolite repressor/activator(Cra)protein of enteric bacteria[J].Journal of Bacteriology,1996,178(12):3411-3417.