Premium
Refolding of lactate dehydrogenase by zeolite beta
Author(s) -
Togashi Hideaki,
Nara Takayuki,
Sekikawa Chisato,
Kawakami Masayuki,
Yaginuma Nakatsugu,
Tsunoda Tatsuo,
Sakaguchi Kengo,
Mizukami Fujio
Publication year - 2009
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.107
Subject(s) - zeolite , chemistry , adsorption , polyethylene glycol , guanidine , peg ratio , lactate dehydrogenase , arginine , nuclear chemistry , yield (engineering) , hydrochloride , chromatography , biochemistry , enzyme , organic chemistry , catalysis , amino acid , materials science , finance , economics , metallurgy
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%. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009