Abstract
Using 2-dimensional gel electrophoresis, the Escherichia coli proteome response to a heat-shock stress was analyzed and a 1.6-fold increase of malate dehydrogenase was observed even under the heat-shock condition where the total number of soluble proteins decreased by about 5%. We subsequently demonstrated that, as an N-terminus fusion expression partner, malate dehydrogenase facilitated the folding of, and dramatically increased the solubility of, many aggregation-prone heterologous proteins in E. coli cytoplasm. Therefore, malate dehydrogenase is well suited for production of a biologically active fusion mutant of cutinase (Pseudomonas putida origin) that is currently of considerable to biotechnology and commercial industries.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahn JY, Choi H, Kim YH, Han KY, Park JS, Han SS, Lee J (2005) Heterologous gene expression using self-assembled supra-molecules with high affinity for HSP70 chaperone. Nucleic Acids Res 33(12):3751–3762
Basha E, Lee GJ, Demeler B, Vierling E (2004) Chaperone activity of cytosolic small heat shock proteins from wheat. Eur J Biochem 271(8):1426–1436
Bedouelle H, Duplay P (1998) Production in Escherichia coli and one-step purification of bifunctional hybrid proteins which bind maltose. Export of the Klenow polymerase into the periplasmic space. Eur J Biochem 71(3):541–549
Braun P, Hu Y, Shen B, Halleck A, Koundinya M, Harlow E, LaBaer J (2002) Proteome-scale purification of human proteins from bacteria. Proc Natl Acad Sci USA 99(5):2654–2659
Dihazi H, Asif AR, Agarwal NK, Doncheva Y, Muller GA (2005) Proteomic analysis of cellular response to osmotic stress in thick ascending limb of Henle’s loop (TALH) cells. Mol Cell Proteomics 4(10):1445–1458
Ding HT, Ren H, Chen Q, Fang G, Li LF, Li R, Wang Z, Jia XY, Liang YH, Hu MH, Li Y, Luo JC, Gu XC, Su XD, Luo M, Lu SY (2002) Parallel cloning, expression, purification and crystallization of human proteins for structural genomics. Acta Crystallogr D Biol Crystallogr 58(Pt 12):2102–2108
Gerard HC, Fett WF, Osman SF, Moreau RA (1993) Evaluation of cutinase activity of various industrial lipases. Biotechnol Appl Biochem 17:181–189
Hammarstrom M, Woestenenk EA, Hellgren N, Hard T, Berglund H (2006) Effect of N-terminal solubility enhancing fusion proteins on yield of purified target protein. J Struct Funct Genomics 7(1):1–14
Himanen JP, Rajashankar KR, Lackmann M, Cowan CA, Henkemeyer M, Nikolov DB (2001) Crystal structure of an Eph receptor-ephrin complex. Nature 414(6866):933–938
Jürgen B, Hanschke R, Sarvas M, Hecker M, Schweder T (2001) Proteome and transcriptome based analysis of Bacillus subtilis cells overproducing an insoluble heterologous protein. Appl Microbiol Biotechnol 55(3):326–332
Kim YH, Han KY, Lee K, Heo JH, Kang HY, Lee J (2004) Comparative proteome analysis of Hansenula polymorpha DL1 and A16. Proteomics 4:2005–2013
Kim YH, Lee J, Moon SH (2003) Uniqueness of microbial cutinase in hydrolysis of p-Nitrophenyl esters. J Microbial Biotechnol 13(1):57–63
Lin T-S, Kolattukudy PE (1980) Structural studies on cutinase, a glycoprotein containing novel amino acids and glucuronic acid amide at the N terminus. Eur J Biochem 106:341–351
Motojima F, Chaudhry C, Fenton WA, Farr GW, Horwich AL (2004) Substrate polypeptide presents a load on the apical domains of the chaperonin GroEL. Proc Natl Acad Sci USA 101(42):15005–150012
Nishihara K, Kanemori M, Kitagawa M, Yanagi H, Yura T (1998) Chaperone coexpression plasmids: differential and synergistic roles of DnaK-DnaJ-GrpE and GroEL-GroES in assisting folding of an allergen of Japanese cedar pollen, Cryj2, in Escherichia coli. Appl Environ Microbiol 64(5):1694–1699
Parsell DA, Sauer RT (1989) Induction of a heat shock-like response by unfolded protein in Escherichia coli: dependence on protein level not protein degradation. Genes Dev 3(8):1226–1232
Rao D, Momcilovic I, Kobayashi S, Callegari E, Ristic Z (2004) Chaperone activity of recombinant maize chloroplast protein synthesis elongation factor, EF-Tu. Eur J Biochem 271(18):3684–3692
Schein CH, Noteborn MHM (1998) Formation of Soluble Recombinant Proteins in Escherichia coli is favored by lower growth temperatures. Biotechnology (NY) 6:291–294
Smith DB, Johnson KS (1988) Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67(1):31–40
Stenström CM, Jin H, Major LL, Tate WP, Isaksson LA (2001) Codon bias at the 3′-side of the initiation codon is correlated with translation initiation efficiency in Escherichia coli. Gene 263(1–2):273–284
Acknowledgements
We thank Professor Hang Chul Shin at Soongsil University for kindly providing the gene clones of mpINS and G-CSF. We also appreciate Professors Won Tae Lee and Hyun Soo Cho at Yonsei University for the kind donation of ppGRN, AID, and Nacht clones, respectively. This study was supported by the Korea Health 21 R&D Project of the Ministry of Health & Welfare of the Republic of Korea (grant no. A050750). This work was also supported by the Second Brain Korea 21 Project. Further supports from the Korea Science and Engineering Foundation (grant no. R01-2005-000-10355-0), the Korea Research Foundation (grant no. KRF-2004-041-D00180), and the Microbial Genomics & Applications Center (Taejon, Republic of Korea) are also appreciated.
Author information
Authors and Affiliations
Corresponding author
Additional information
Jin-Seung Park and Kyung-Yeon Han contributed equally to this work.
Rights and permissions
About this article
Cite this article
Park, JS., Han, KY., Song, JA. et al. Escherichia coli malate dehydrogenase, a novel solubility enhancer for heterologous proteins synthesized in Escherichia coli . Biotechnol Lett 29, 1513–1518 (2007). https://doi.org/10.1007/s10529-007-9417-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-007-9417-3