Abstract
Objectives
Bacillus licheniformis WX-02 is used for the production of many valuable chemicals. Here, we have sought to improve l-valine production by blocking the metabolic pathways related to branched-chain amino acids.
Results
The synthesis genes of l-leucine (leuA) and l-isoleucine (ilvA) were deleted to obtain mutant strains. l-Valine yields of WX-02ΔleuA and WX-02ΔilvA reached 33.2 and 21.1 mmol/l, respectively, which are 22 and 14 times higher than the wild-type WX-02 (1.53 mmol/l). After further deletion of l-lactate dehydrogenase gene (ldh) from WX-02ΔleuA, the productivity reached 0.47 mmol/l h, an increase of 19 %.
Conclusion
We provide a possibility to over-produce l-valine using genetically-modified B. licheniformis using remodeling of the biosynthetic pathway to l-valine.
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References
Bartek T, Makus P, Klein B, Lang S, Oldiges M (2008) Influence of l-isoleucine and pantothenate auxotrophy for l-valine formation in Corynebacterium glutamicum revisited by metabolome analyses. Bioprocess Biosyst Eng 31:217–225
Bartek T, Blombach B, Zönnchen E, Makus P, Lang S, Eikmanns BJ, Oldiges M (2010) Importance of NADPH supply for improved l-valine formation in Corynebacterium glutamicum. Biotechnol Prog 26:361–371
Blombach B, Schreiner ME, Holatko J (2007) l-Valine production with pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum. Appl Environ Microbiol 73:2079–2084
Blombach B, Schreiner ME, Bartek T, Oldiges M, Eikmanns BJ (2008) Corynebacterium glutamicum tailored for high-yield l-valine production. Appl Microbiol Biotechnol 79:471–479
Chattopadhyay SP, Banerjee AK (1978) Production of valine by a Bacillus sp. Z Allg Mikrobiol 18:243–254
Iborra JL, Obon JM, Manjon A, Canovas M (1992) Analysis of a laminated enzyme membrane reactor for continuous resolution of amino acids. Biotechnol Appl Biochem 15:22–30
Khachatryan AG (1986) Dependence of valine production by serratia marcescens on the ion composition of the fermentation. Microbiology 22:554–556
Mudiam MK, Ratnasekhar Ch (2013) Ultra sound assisted one step rapid derivatization and dispersive liquid–liquid microextraction followed by gas chromatography–mass spectrometric determination of amino acids in complex matrices. J Chromatogr A 1291:10–18
Park JH, Lee KH, Kim TY, Lee SY (2007) Metabolic engineering of Escherichia coli for the production of l-valine based on transcriptome analysis and in silico gene knockout simulation. Proc Natl Acad Sci 104:7797–7802
Qi G, Kang Y, Li L, Xiao A, Zhang S, Wen Z, Xu D, Chen S (2014) Deletion of meso-2,3-butanediol dehydrogenase gene budC for enhanced d-2,3-butanediol production in Bacillus licheniformis. Biotechnol Biofuels 7(1):16
Radmacher E, Vaitsikova A, Burger U, Krumbach K, Sahm H, Eggeling L (2002) Linking central metabolism with increased pathway flux: l-valine accumulation by Corynebacterium glutamicum. Appl Environ Microbiol 68(5):2246–2250
Santos CC, Libeck BD, Schwan RF (2014) Co-culture fermentation of peanut-soy milk for the development of a novel functional beverage. Int J Food Microbiol 186(1):32–41
Sohail M (1998) A simple and rapid method for preparing genomic DNA from gram-positive bacteria. Mol Biotechnol 10(2):191–193
Wang Q, Chen T, Zhao X, Chamu J (2012) Metabolic engineering of thermophilic Bacillus licheniformis for chiral pure d-2,3-butanediol production. Biotechnol Bioeng 109:1610–1621
Wei X, Ji Z, Chen S (2010) Isolation of halotolerant Bacillus licheniformis WX-02 and regulatory effects of sodium chloride on yield and molecular sizes of poly-γ-glutamic acid. Appl Biochem Biotechnol 160(5):1332–1340
Xue G, Johnson JS, Dalrymple BP (1999) High osmolarity improves the electro-transformation efficiency of the gram-positive bacteria Bacillus subtilis and Bacillus licheniformis. J Microbiol Methods 34(3):183–191
Yangtse W, Zhou Y, Lei Y, Qiu Y, Wei X, Ji Z, Qi G, Yong Y, Chen L, Chen S (2012) Genome sequence of Bacillus licheniformis WX-02. J Bacteriol 194(13):3561–3562
Acknowledgments
This work was supported by National Program on Key Basic Research Project (973 Program, No. 2015CB150505).
Supporting Information
Supplementary Table 1 – Primers used for PCR in this study.
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Liang, C., Huo, Y., Qi, G. et al. Enhancement of l-valine production in Bacillus licheniformis by blocking three branched pathways. Biotechnol Lett 37, 1243–1248 (2015). https://doi.org/10.1007/s10529-015-1783-7
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DOI: https://doi.org/10.1007/s10529-015-1783-7