Refolding of lactate dehydrogenase by zeolite beta

Autor:	Masayuki Kawakami, Fujio Mizukami, Tatsuo Tsunoda, Hideaki Togashi, Chisato Sekikawa, Nakatsugu Yaginuma, Takayuki Y. Nara, Kengo Sakaguchi
Rok vydání:	2009
Předmět:	Protein Folding Chromatography L-Lactate Dehydrogenase Arginine Chemistry Hydrochloride Polyethylene glycol Recombinant Proteins chemistry.chemical_compound Adsorption Lactate dehydrogenase PEG ratio Zeolites Zeolite Guanidine Biotechnology
Zdroj:	Biotechnology Progress. 25:200-206
ISSN:	1520-6033 8756-7938
Popis:	We used zeolite beta as an adsorbing matrix to refold recombinant lactate dehydrogenase (LDH) protein collected as an insoluble aggregate from a bacterial expression system. The adsorption isotherm revealed that 1 g of zeolite adsorbed 200 mg of denatured LDH solubilized with a buffer containing 6 M of guanidine hydrochloride. The pH of the buffer had little effect on the adsorption, but this property was abolished by preincubation of the zeolite with polyethylene glycol (PEG) in a weight ratio of 1:10. These data suggest that the adsorption of LDH depends on the hydrophobicity of the zeolite surface, and that the adsorption of PEG to zeolite is sufficient to release LDH from its surface. LDH was thus released by refolding buffer containing PEG and arginine, and soluble LDH was obtained in its active enzymatic form. The addition of arginine dramatically increased the yield of LDH in a dose-dependent manner. The overall refolding efficiency was optimized to 35%.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f5f5350b7cb3f30af11479bfaa0f4d94 https://doi.org/10.1002/btpr.107 Zobrazit plný text záznamu Plný text