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Low Concentration of Salt Triggers Elastin-like Polypeptides Inverse Transition to Purify 1,3-propanediol Oxidoreductase(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2013 04
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Low Concentration of Salt Triggers Elastin-like Polypeptides Inverse Transition to Purify 1,3-propanediol Oxidoreductase
GE Huihua HUANG Zhihong WANG Wenyan ZHANG Shiran ZHANG Guangya
(Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China)
elastin-like polypeptides purification tag inverse temperature transition recombinant enzyme purification non-chromatographic

Elastin-like polypeptides have been widely used as the purification tag. Non-chromatographic purification of ELPs fusion proteins, termed inverse transition cycling (ITC), exploits the reversible soluble-insoluble phase transition behavior imparted by the ELPs tag. This study found that the novel ELPs tag could purify 1,3-propanediol oxidoreductase (PDOR) efficiently. The ELPs aggregated with 0.8 mol/L Na2CO3 at room temperature, and then PDOR, which was linked with the ELPs, was purified by ITC. The purity could reach about 98%, and the purification fold was above 8, about 7 times that of His-tag purification method. By comparing the properties of the purified fusion ELPs-PDOR and His-PDOR, we found that ELPs tag hardly affects the target protein, suggesting that it can be used during the biocatalysis process. The method with low salt concentration to induce the phase transition of ELPs can avoid potential adverse effect on enzyme structure and property, thus being advantageous in protein purification. Fig 3, Tab 2, Ref 17


Urry DW. Physical chemistry of biological free energy transduction as demonstrated by elastic protein-based polymers [J]. Phys Chem, 1997, 51: 11007-11028
Urry DW. Free energy transduction in polypeptides and proteins based on inverse temperature transitions [J]. Prog Biophys Mol Biol, 1992, 57: 23-57
Lim DW, Kimberly TC, MacKay JA, Chilkoti A. Improved non-chromatographic purification of a recombinant protein by cationic elastin-like polypeptides [J]. Biomacromolecules, 2007, 8 (5): 1417-1424
Floss DM, Schallau K, Rose-John S, Conrad U, Scheller J. Elastin-like polypeptides revolutionize recombinant protein expression and their biomedical application [J]. Trends Biotechnol, 2010, 28: 37-45
Trabbic-Carlson K, Liu L, Kim B, Chilkoti A. Expression and purification of recombinant proteins from Escherichia coli: comparison of an elastin-like polypeptide fusion with an oligohistidine fusion [J]. Protein Sci, 2004, 13: 3274-3284
Banki MR, Feng L, Wood DW. Simple bioseparations using self-cleaving elastin-like polypeptide tags [J]. Nat Methods, 2005, 2: 659-661
Baley AF, Wu WY, David WW. Optimization of ELP-intein mediated protein purifcation by salt substitution [J]. Protein Express Purif, 2009, 66: 198-202
Chow D, Nunalee ML, Lim DW, Simnick AJ, Chilkoti A. Peptide-based biopolymers in biomedicine and biotechnology [J]. Mater Sci Eng Rep, 2008, 62: 125-155
黄凯宗, 李晶晶, 王文研, 李巍, 张光亚. 类弹性蛋白多肽的从头设计、非色谱纯化及盐效应[J]. 生物工程学报, 2011, 27 (4): 661-666 [Huang KZ, Li JJ, Wang WY, Li W, Zhang GY. De novo design, non-chromatographic purification and salts effect of elastin-like polypeptides [J]. Chin J Biotech, 2011, 27 (4): 661-666]
Li W, Ng IS, Fang BS, Zhang GY. Codon optimization of 1,3-propanediol oxidoreductase expression in Escherichia coli and enzymatic properties [J]. Electron J Biotechnol, 2011, 14 (4), DOI: 10.2225/vol14-issue4-fulltext-9
Ahrens K, Menzel K, Zeng A, Deckwer W. Kinetic, dynamic, and pathway studies of glycerol metabolism by Klebsiella pneumoniae in anaerobic continuous culture. III. Enzymes and ?uxes of glycerol dissimilation and 1,3-propanediol formation [J]. Biotechnol Bioeng, 1998, 59: 544-552
陈宏文, 聂金峰, 方柏山. 克雷伯杆菌1,3-丙二醇氧化还原酶的分离纯化及其酶学性质[J]. 华侨大学报(自然科学版), 2009, 30: 62-66 [Chen HW, Nie JF, Fang BS. Purification and characterization of 1,3-propanediol oxidoreductase from Klebsiella pneumonia [J]. J Huaqiao Univ (Nat Sci), 2009, 30: 62-66]
Cho Y, Zhang Y, Christensen T, Sagle LB, Chilkoti A, Cremer PS. Effects of Hofmeister anions on the phase transition temperature of elastin-like polypeptides [J]. Phys Chem B, 2008, 112 (44): 13765-13771
Ge X, Yang DS, Trabbic-Carlson K, Kim B, Chilkoti A, Filipe CD. Self-cleavable stimulus responsive tags for protein purification without chromatography [J]. Am Chem Soc, 2005, 127: 11228-11229
Meyer DE, Chilkoti A. Genetically encoded synthesis of protein-based polymers with precisely specified molecular weight and sequence by recursive directional ligation: examples from the elastin-like polypeptide system [J]. Biomacromolecules, 2002, 3: 357-367
付晓平, 王文研, 张光亚. 以类弹性蛋白多肽为标签的表达质粒构建及其用于木聚糖酶的非色谱纯化 [J]. 微生物学报, 2012, 52 (1): 90-95 [Fu XP, Wang WY, Zhang GY. Construction of an expression vector with elastin-like polypeptide tag to purify xylanase [J]. Acta Microbiol Sin, 2012, 52 (1): 90-95]
方慧英, 张成, 诸葛斌, 诸葛健. 产1,3-丙二醇新型基因工程菌的构建[J]. 应用与环境生物学报, 2009, 15 (5): 708-712 [Fang HY, Zhang C, Zhuge B, Zhuge J. Construction of novel recombinant strains capable of producing 1,3-propanediol [J]. Chin J Appl Environ Biol, 2009, 15 (5): 708-712]


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