Metabolic Engineering of Carbohydrate Metabolism Systems in Corynebacterium Glutamicum for Improving the Efficiency of L-lysine Production from Mixed Sugar

Overview

Journal Microb Cell Fact

Publisher Biomed Central

Specialties Biotechnology
Microbiology

Date 2020 Feb 20

PMID 32070345

Citations 10

Authors

Jian-Zhong Xu

Hao-Zhe Ruan

Hai-Bo Yu

Li-Ming Liu

Weiguo Zhang

Affiliations

Soon will be listed here.

Abstract

The efficiency of industrial fermentation process mainly depends on carbon yield, final titer and productivity. To improve the efficiency of L-lysine production from mixed sugar, we engineered carbohydrate metabolism systems to enhance the effective use of sugar in this study. A functional metabolic pathway of sucrose and fructose was engineered through introduction of fructokinase from Clostridium acetobutylicum. L-lysine production was further increased through replacement of phosphoenolpyruvate-dependent glucose and fructose uptake system (PTS and PTS) by inositol permeases (IolT1 and IolT2) and ATP-dependent glucokinase (ATP-GlK). However, the shortage of intracellular ATP has a significantly negative impact on sugar consumption rate, cell growth and L-lysine production. To overcome this defect, the recombinant strain was modified to co-express bifunctional ADP-dependent glucokinase (ADP-GlK/PFK) and NADH dehydrogenase (NDH-2) as well as to inactivate SigmaH factor (SigH), thus reducing the consumption of ATP and increasing ATP regeneration. Combination of these genetic modifications resulted in an engineered C. glutamicum strain K-8 capable of producing 221.3 ± 17.6 g/L L-lysine with productivity of 5.53 g/L/h and carbon yield of 0.71 g/g glucose in fed-batch fermentation. As far as we know, this is the best efficiency of L-lysine production from mixed sugar. This is also the first report for improving the efficiency of L-lysine production by systematic modification of carbohydrate metabolism systems.

Citing Articles

Reconstruction the feedback regulation of amino acid metabolism to develop a non-auxotrophic L-threonine producing Corynebacterium glutamicum.

Liu J, Liu J, Li J, Zhao X, Sun G, Qiao Q Bioresour Bioprocess. 2024; 11(1):43.

PMID: 38664309 PMC: 11045695. DOI: 10.1186/s40643-024-00753-9.

Enhanced L-ornithine production from glucose and sucrose via manipulation of the fructose metabolic pathway in Corynebacterium glutamicum.

Nie L, Xu K, Zhong B, Wu X, Ding Z, Chen X Bioresour Bioprocess. 2024; 9(1):11.

PMID: 38647759 PMC: 10992749. DOI: 10.1186/s40643-022-00503-9.

Metabolic flux analysis: a comprehensive review on sample preparation, analytical techniques, data analysis, computational modelling, and main application areas.

de Falco B, Giannino F, Carteni F, Mazzoleni S, Kim D RSC Adv. 2022; 12(39):25528-25548.

PMID: 36199351 PMC: 9449821. DOI: 10.1039/d2ra03326g.

Transcriptome profiles of high-lysine adaptation reveal insights into osmotic stress response in .

Wang J, Yang J, Shi G, Li W, Ju Y, Wei L Front Bioeng Biotechnol. 2022; 10:933325.

PMID: 36017356 PMC: 9395588. DOI: 10.3389/fbioe.2022.933325.

Strategy for the design of a bioreactor for L-lysine immobilized fermentation using Corynebacterium glutamicum.

Meng G, Zhao W, Liu Q, Zhao C, Zou Y, Zhao N Appl Microbiol Biotechnol. 2022; 106(17):5449-5458.

PMID: 35902409 DOI: 10.1007/s00253-022-12103-w.

References

Sagong H, Kim K . Structural basis for redox sensitivity in Corynebacterium glutamicum diaminopimelate epimerase: an enzyme involved in l-lysine biosynthesis. Sci Rep. 2017; 7:42318. PMC: 5296763. DOI: 10.1038/srep42318. View

Xu J, Wu Z, Gao S, Zhang W . Rational modification of tricarboxylic acid cycle for improving L-lysine production in Corynebacterium glutamicum. Microb Cell Fact. 2018; 17(1):105. PMC: 6035423. DOI: 10.1186/s12934-018-0958-z. View

Hoffmann S, Jungmann L, Schiefelbein S, Peyriga L, Cahoreau E, Portais J . Lysine production from the sugar alcohol mannitol: Design of the cell factory Corynebacterium glutamicum SEA-3 through integrated analysis and engineering of metabolic pathway fluxes. Metab Eng. 2018; 47:475-487. DOI: 10.1016/j.ymben.2018.04.019. View

Krings E, Krumbach K, Bathe B, Kelle R, Wendisch V, Sahm H . Characterization of myo-inositol utilization by Corynebacterium glutamicum: the stimulon, identification of transporters, and influence on L-lysine formation. J Bacteriol. 2006; 188(23):8054-61. PMC: 1698185. DOI: 10.1128/JB.00935-06. View

Ikeda M . Sugar transport systems in Corynebacterium glutamicum: features and applications to strain development. Appl Microbiol Biotechnol. 2012; 96(5):1191-200. DOI: 10.1007/s00253-012-4488-z. View

Yin F, Zhu S, Guo D, Ren L, Ji X, Huang H . Development of a strategy for the production of docosahexaenoic acid by Schizochytrium sp. from cane molasses and algae-residue. Bioresour Technol. 2018; 271:118-124. DOI: 10.1016/j.biortech.2018.09.114. View

Becker J, Zelder O, Hafner S, Schroder H, Wittmann C . From zero to hero--design-based systems metabolic engineering of Corynebacterium glutamicum for L-lysine production. Metab Eng. 2011; 13(2):159-68. DOI: 10.1016/j.ymben.2011.01.003. View

Richter J, Goroncy A, Ronimus R, Sutherland-Smith A . The Structural and Functional Characterization of Mammalian ADP-dependent Glucokinase. J Biol Chem. 2015; 291(8):3694-704. PMC: 4759153. DOI: 10.1074/jbc.M115.679902. View

Barriuso-Iglesias M, Barreiro C, Flechoso F, Martin J . Transcriptional analysis of the F0F1 ATPase operon of Corynebacterium glutamicum ATCC 13032 reveals strong induction by alkaline pH. Microbiology (Reading). 2005; 152(Pt 1):11-21. DOI: 10.1099/mic.0.28383-0. View

10.

Becker J, Klopprogge C, Zelder O, Heinzle E, Wittmann C . Amplified expression of fructose 1,6-bisphosphatase in Corynebacterium glutamicum increases in vivo flux through the pentose phosphate pathway and lysine production on different carbon sources. Appl Environ Microbiol. 2005; 71(12):8587-96. PMC: 1317465. DOI: 10.1128/AEM.71.12.8587-8596.2005. View

11.

Baumchen C, Krings E, Bringer S, Eggeling L, Sahm H . Myo-inositol facilitators IolT1 and IolT2 enhance D-mannitol formation from D-fructose in Corynebacterium glutamicum. FEMS Microbiol Lett. 2008; 290(2):227-35. DOI: 10.1111/j.1574-6968.2008.01425.x. View

12.

Xu J, Yang H, Zhang W . NADPH metabolism: a survey of its theoretical characteristics and manipulation strategies in amino acid biosynthesis. Crit Rev Biotechnol. 2018; 38(7):1061-1076. DOI: 10.1080/07388551.2018.1437387. View

13.

Brusseler C, Radek A, Tenhaef N, Krumbach K, Noack S, Marienhagen J . The myo-inositol/proton symporter IolT1 contributes to d-xylose uptake in Corynebacterium glutamicum. Bioresour Technol. 2017; 249:953-961. DOI: 10.1016/j.biortech.2017.10.098. View

14.

Castro-Fernandez V, Bravo-Moraga F, Herrera-Morande A, Guixe V . Bifunctional ADP-dependent phosphofructokinase/glucokinase activity in the order Methanococcales--biochemical characterization of the mesophilic enzyme from Methanococcus maripaludis. FEBS J. 2014; 281(8):2017-29. DOI: 10.1111/febs.12757. View

15.

Bott M, Niebisch A . The respiratory chain of Corynebacterium glutamicum. J Biotechnol. 2003; 104(1-3):129-53. DOI: 10.1016/s0168-1656(03)00144-5. View

16.

Wu W, Zhang Y, Liu D, Chen Z . Efficient mining of natural NADH-utilizing dehydrogenases enables systematic cofactor engineering of lysine synthesis pathway of Corynebacterium glutamicum. Metab Eng. 2018; 52:77-86. DOI: 10.1016/j.ymben.2018.11.006. View

17.

Chen Z, Huang J, Wu Y, Wu W, Zhang Y, Liu D . Metabolic engineering of Corynebacterium glutamicum for the production of 3-hydroxypropionic acid from glucose and xylose. Metab Eng. 2016; 39:151-158. DOI: 10.1016/j.ymben.2016.11.009. View

18.

Nath S . Two-ion theory of energy coupling in ATP synthesis rectifies a fundamental flaw in the governing equations of the chemiosmotic theory. Biophys Chem. 2017; 230:45-52. DOI: 10.1016/j.bpc.2017.08.005. View

19.

Kengen S, de Bok F, van Loo N, Dijkema C, Stams A, de Vos W . Evidence for the operation of a novel Embden-Meyerhof pathway that involves ADP-dependent kinases during sugar fermentation by Pyrococcus furiosus. J Biol Chem. 1994; 269(26):17537-41. View

20.

Nantapong N, Kugimiya Y, Toyama H, Adachi O, Matsushita K . Effect of NADH dehydrogenase-disruption and over-expression on respiration-related metabolism in Corynebacterium glutamicum KY9714. Appl Microbiol Biotechnol. 2004; 66(2):187-93. DOI: 10.1007/s00253-004-1659-6. View