Abstract
A new phosphoenolpyruvate carboxylase (PEPC) gene of Dunaliella salina is identified using homology analysis was conducted using PEPC gene of Chlamydomonas reinhardtii and Arabidopsis thaliana. Recombinant E. coli SGJS115 with increased production of malate and oxaloacetate was developed by introducing codon-optimized phosphoenolpyruvate carboxylase2 (OPDSPEPC2) gene of Dunaliella salina. E. coli SGJS115 yielded a 9.9 % increase in malate production. In addition, E. coli SGJS115 exhibited two times increase in the yield of oxaloacetate over the E. coli SGJS114 having identified PEPC2 gene obtained from Dunaliella salina.
References
Rintze MZ, Erik DH, Wouter AW, Pieter DW, Cor D, Aaron AW, Maarten AG, Johannes PD, Jack TP, Antonius JAM (2008) Malic acid production by Saccharomyces cerevisiae: engineering of pyruvate carboxylation, oxaloacetate reduction, and malate export. Appl Environ Microbiol 74:2766–2777
Battat E, Peleg Y, Bercovitz A, Rokem JS, Goldberg I (1991) Optimization of l-malic acid production by Aspergillus flavus in a stirred fermentor. Biotechnol Bioeng 37:1108–1116
Kwon YD, Kwon OH, Lee SH, Kim P (2007) The effect of NADP-dependent malic enzyme expression and anaerobic C4 metabolism in Escherichia coli compared with other anaplerotic enzymes. J Appl Microbiol 103:2340–2345
Millard CS, Chao YP, Liao JC, Donnelly MI (1996) Enhanced production of succinic acid by overexpression of phosphoenolpyruvate carboxylase in Escherichia coli. Appl Environ Microbiol 62:1808–1810
Rivoal J, Dunford R, William CP, David HT (1996) Purification and properties of four phosphoenolpyruvate carboxylase isoforms from the green alga Selenastrum minutum: evidence that association of the 102-kDa catalytic subunit with unrelated polypeptides may modify the physical and kinetic properties of the enzyme. Arch Biochem Biophysics 332:47–57
Kodaki T, Katagiri F, Asano M, Izui K, Katsuki H (1985) Cloning of phosphoenolpyruvate carboxylase gene from a Cyanobacterium Anacystis nidulans in Escherichia coli. J Biochem 97:533–539
Giordano M, Pezzoni V, Hell R (2000) Strategies for the allocation of resources under sulfur limitation in the green alga Dunaliella salina. Plant Physiol 124:857–864
Corpet F (1998) Multiple sequence alignment with hierarchical clustering. Nucl Acids Res 16:10881–10890
Brown NA, Sharma S, Sobott F, Loenarz C, Oppermann U, Gileadi O (2008) Codon optimization can improve expression of human genes in Escherichia coli: a multi-gene study. Protein Expr Purif 59:94–102
Feng Z, Zhang L, Han X, Zhang Y (2010) Codon optimization of the calf prochymosin gene and its expression in Kluyveromyces lactis. World J Microbiol Biotechnol 26:898–901
Peter KM, Philippe R (1998) Role of phosphoenolpyruvate carboxylase in malate production by the developing barley aleurone layer. Physiol Plantarum 103:132–138
Yoon SH, Lee SH, Das A, Ryu HK, Jang HJ, Kim JY, Oh DK, Keasling JD, Kim SW (2009) Combinatorial expression of bacterial whole mevalonate pathway for the production of β-carotene in E. coli. J Biotechnol 140:218–226
Acknowledgments
This work was supported by the National Research Foundation of Korea Grant funded by the Korean Government (MEST) (NRF-C1ABA001-2010-0020501).
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Park, S., Chang, K.S., Jin, E. et al. Oxaloacetate and malate production in engineered Escherichia coli by expression of codon-optimized phosphoenolpyruvate carboxylase2 gene from Dunaliella salina . Bioprocess Biosyst Eng 36, 127–131 (2013). https://doi.org/10.1007/s00449-012-0759-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-012-0759-4