Optimization of DsbA Purification from Recombinant Escherichia coli Broth Using Box-Behnken Design Methodology

doi:10.1016/S1004-9541(13)60457-7

Abstract

Abstract: Disulfide bond formation protein A (DsbA) is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was (377.9?1.7) mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gi|2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro.

Key words: disulfide bond formation protein A, protein purification, Box-Behnken experiment design, response surface methodology, multi-object programming

摘要： Disulfide bond formation protein A (DsbA) is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was (377.9?1.7) mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gi|2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro.

关键词: disulfide bond formation protein A, protein purification, Box-Behnken experiment design, response surface methodology, multi-object programming

LUO Man, GUAN Yixin, YAO Shanjing. Optimization of DsbA Purification from Recombinant Escherichia coli Broth Using Box-Behnken Design Methodology[J]. Chin.J.Chem.Eng., 2013, 21(2): 185-191.

罗曼, 关怡新, 姚善泾. Optimization of DsbA Purification from Recombinant Escherichia coli Broth Using Box-Behnken Design Methodology[J]. Chinese Journal of Chemical Engineering, 2013, 21(2): 185-191.

References

1 Bardwell, J.C.A., McGovern, K., Beckwith, J., “Identification of a protein required for disulfide bond formation in vivo”, Cell, 67, 581-589 (1991).

2 Kamitani, S., Akiyama, Y., Ito, K., “Identification and characterization of an Escherichia coli gene required for the formation of correctly folded alkaline phosphatase, a periplasmic enzyme”, EMBO J, 11, 57-62 (1992).

3 Plunkett, G., Burland, V., Daniels, D.L., Blattner, F.R., “Analysis of the Escherichia coli genome. III. DNA sequence of the region from 87.2 to 89.2 minutes”, Nucleic Acids Res., 21, 3391-3398 (1993).

4 Wunderlich, M., Jaenicke, R., Glockshuber, R., “The redox properties of protein disulfide isomerase (DsbA) of Eshcherichia coli result from a tense conformation of its oxidized form”, J. Mol. Biol., 233, 559-566 (1993).

5 Zheng, W.D., Quan, H., Song, J.L., Yang, S.L., Wang, C.C., “Does DsbA have chaperone-like activity?”, Arch. Biochem. Biophys., 337, 326-331 (1997).

6 Debarbieux, L., Beckwith, J., “Electron avenue: Pathways of disulfide bond formation and isomerization”, Cell, 99, 117-119 (1999).

7 Collet, J.F., Bardwell, J.C.A., “Oxidative protein folding in bacteria”, Mol. Microbiol., 44, 1-8 (2002).

8 Box, G., Behnken, D., “Some new three-level designs for the study of quantitative variables”, Technometrics, 2, 455-475 (1960).

9 Ferreira, S.L.C., Bruns, R.E., Ferreira, H.S., Matos, G.D., David, J.M., Brandao, G.C., da Silva, E.G.P., Portugal, L.A., dos Reis, P.S., Souza, A.S., dos Santos, W.N.L., “Box-Behnken design: An alternative for the optimization of analytical methods”, Anal. Chim. Acta, 597, 179-186 (2007).

10 Sun, M., “Some issues in measuring and reporting solution quality of interactive multiple objective programming procedures”, Eur. J. Oper. Res., 162, 468-483 (2005).

11 Tozer, P.R., Stokes, J.R. “A multi-objective programming approach to feed ration balancing and nutrient management”, Agr. Syst., 67, 205-215 (2001).

12 Sun, M., “A multiple objective programming approach for determining faculty salary equity adjustments”, Eur. J. Oper. Res., 138, 302-319 (2002).

13 Pappin, D.J.C., Hojrup, P., Bleasby, A.J., “Rapid identification of proteins by peptide-mass fingerprinting”, Curr. Biol., 3, 327-332 (1993).

14 Mann, M., Højrup, P., Roepstorff, P., “Use of mass spectrometric molecular weight information to identify proteins in sequence databases”, Biol. Mass Spectrom., 22, 338-345 (1993).

15 Henzela, W.J., Watanabea, C., Stults, J.T., “Protein identification: the origins of peptide mass fingerprinting”, J. Am. Soc. Mass Spectr., 14, 931-942 (2003).

16 Luo, M., Guan, Y.X., Yao, S.J., “Statistical optimization of protein disulfide isomerase expression in E. coli”, Chin. J. Biochem. Mol. Biol., 21, 376-383 (2005).

17 Zhang, C., Fan, D.D., Shang, L.A., Ma, X.X., Luo, Y.E., Xue, W.J., Gao, P.F., “Optimization of fermentation process for human-like collagen production of recombinant Escherichia coli using response surface methodology”, Chin. J. Chem. Eng., 18, 137-142 (2010).

18 Bradford, M.M., “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding”, Anal. Biochem., 72, 248-254 (1976).

19 Stellmach, B., Qian, J.Y., Enzyme Measurement, China Light Industry Press, Beijing (1992). (in Chinese)

20 Luo, M., Guan, Y.X., Yao, S.J., “Study on disulfide bond formation protein A in Escherichia coli”, Chin. J. Biotech., 23 (1), 7-15 (2007).

[1]	Kamel Hendaoui, Malika Trabelsi-Ayadi, Fadhila Ayari. Optimization of continuous electrocoagulation-adsorption combined process for the treatment of a textile effluent [J]. Chinese Journal of Chemical Engineering, 2022, 44(4): 310-320.
[2]	Zenan Wang, Xin Zheng, Yan Wang, Heng Lin, Hui Zhang. Evaluation of phenanthrene removal from soil washing effluent by activated carbon adsorption using response surface methodology [J]. Chinese Journal of Chemical Engineering, 2022, 42(2): 399-405.
[3]	Qingjun Zhang, Youguang Ma, Xigang Yuan, Aiwu Zeng. Box-Behnken experimental design for optimizing process parameters in carbonate-promoted direct thiophene carboxylation reaction with carbon dioxide [J]. Chinese Journal of Chemical Engineering, 2022, 50(10): 222-234.
[4]	Kamel Hendaoui, Malika Trabelsi-Ayadi, Fadhila Ayari. Optimization and mechanisms analysis of indigo dye removal using continuous electrocoagulation [J]. Chinese Journal of Chemical Engineering, 2021, 29(1): 242-252.
[5]	Xin Yuan, Lujun Wang, Panliang Zhang, Weifeng Xu, Kewen Tang. Enantioselective esterification of (R,S)-2-(4-methylphenyl) propionic acid via Novozym 435: Optimization and application [J]. Chinese Journal of Chemical Engineering, 2020, 28(7): 1816-1823.
[6]	Mohammad Dana, Mohammad Amin Sobati, Shahrokh Shahhosseini, Aminreza Ansari. Optimization of a continuous ultrasound assisted oxidative desulfurization (UAOD) process of diesel using response surface methodology (RSM) considering operating cost [J]. Chinese Journal of Chemical Engineering, 2020, 28(5): 1384-1396.
[7]	Mohammed Abdullah Issa, Zurina Zainal Abidin, Shafreeza Sobri, Suraya Abdul-Rashid, Mohd Adzir Mahdi, Nor Azowa Ibrahim, Musa Y. Pudza. Fabrication, characterization and response surface method optimization for quantum efficiency of fluorescent nitrogen-doped carbon dots obtained from carboxymethylcellulose of oil palms empty fruit bunch [J]. Chinese Journal of Chemical Engineering, 2020, 28(2): 584-592.
[8]	Huisheng Lü, Jinyi Zhou, Jiatao Liu, Chunliu Lü, Feng Lian, Yonghui Li. Optimization of hydrothermal pretreatment for co-utilization of xylose and glucose of cassava anaerobic residue for producing ethanol [J]. Chinese Journal of Chemical Engineering, 2019, 27(4): 920-927.
[9]	Mohammad Bagher Sabeti, Mohammad Amin Hejazi, Afzal Karimi. Enhanced removal of nitrate and phosphate from wastewater by Chlorella vulgaris: Multi-objective optimization and CFD simulation [J]. Chinese Journal of Chemical Engineering, 2019, 27(3): 639-648.
[10]	Zhijuan Niu, Sitao Zhang, Yanhe Han, Mengmeng Qi. Optimization of the N₂ generation selectivity in aqueous nitrate reduction using internal circulation micro-electrolysis [J]. Chinese Journal of Chemical Engineering, 2019, 27(12): 3010-3016.
[11]	Kangning Xiong, Yun Chen, Shuai Shen. Experimental optimization and mathematical modeling of supercritical carbon dioxide extraction of essential oil from Pogostemon cablin [J]. Chinese Journal of Chemical Engineering, 2019, 27(10): 2407-2417.
[12]	Cuicui Lü, Yongliang Wang, Peng Qian, Ya Liu, Guoyan Fu, Jian Ding, Shufeng Ye, Yuanfa Chen. Separation of chalcopyrite and pyrite from a copper tailing by ammonium humate [J]. Chin.J.Chem.Eng., 2018, 26(9): 1814-1821.
[13]	Min Xia, Chunsong Ye, Kewu Pi, Defu Liu, Andrea R. Gerson. Cr(Ⅲ) removal from simulated solution using hydrous magnesium oxide coated fly ash: Optimization by response surface methodology (RSM) [J]. Chin.J.Chem.Eng., 2018, 26(5): 1192-1199.
[14]	Jiangnan Shen, Zhendong Hou, Congjie Gao. Using bipolar membrane electrodialysis to synthesize di-quaternary ammonium hydroxide and optimization design by response surface methodology [J]. , 2017, 25(9): 1176-1181.
[15]	Hadiseh Khalilpourmeymandi, Azadeh Mirvakili, Mohammad Reza Rahimpour, Alireza Shariati. Application of response surface methodology for optimization of purge gas recycling to an industrial reactor for conversion of CO₂ to methanol [J]. , 2017, 25(5): 676-687.