Abstract
A droplet gel-entrapping method used for enzyme immobilization was improved to simplify the procedure and to increase the enzyme stability. This immobilization technique is suitable for coupled enzyme reactions requiring cofactors. Leucine dehydrogenase (LeuDH) and formate dehydrogenase (FDH) were freeze-dried with bovine serum albumin, dextrin and stabilizers. The freeze-dried enzyme powder was suspended in a methylcellosolve solution containing poly- ethyleneglycol (# 4000) diacrylate, N,N’-methylenebisacrylamide and 2-hydroxyethylacrylate, and the suspension was gelled with initiators. The gel was cut up and the pieces were washed in a buffer to remove the methylcellosolve and the dextrin inside. The maximum conversion ratio for a LeuDH-FDH gel column was determined to be 99.8% by means of the recycling reaction. On long-term operation at 30 °C for leucine production, the initial conversion ratio (7.2%) gradually decreased to 6.6% over the first 10 days. However, the conversion ratio remained almost constant after the 10th day. The effects of flow rate, temperature, pH, and the concentrations of formate, α-ketoisocaproate, ammonium and NAD on the leucine productivity with the gel column were also investigated.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Kajiwara, S. and Maeda, H.: Biotech. Bioeng. 28, 1794 (1986)
Soda, K., Misono, H., Mori, K. and Sakato, H.: Biochem. Biophys. Res. Commun. 44, 931 (1971)
Ohsima, T., Misono, H. and Soda, K.: J. Biol. Chem. 253, 5719 (1987)
Schütte, H., Hummel, W., Tsai, H. and Kula, M.-R.: Appl. Microb. Biotech., 22, 306 (1985)
Ishida, Y., Fujita, T. and Asai, K.: J. Chromatogr. 204, 143 (1981)
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J.: J. Biol. Chem. 193, 265 (1951)
Maeda, H. and Suzuki, H.: Agric. Biol. Chem. 36, 1581 (1972)
Miyawaki, O., Nakamura, K. and Yano, T.: J. Chem. Eng. Japan 15, 142 (1982)
Miyawaki, O., Nakamura, K. and Yano, T.: J. Chem. Eng. Japan 15, 224 (1982)
Dinelli, D., Marconi, W. and Morisi, F.: “Method in Enzymology”, Vol. XLIV, (K. Mosbach, ed.) p. 227, Academic Press, New York 1976
Nakamura, K., Hibino, K. and Yano, T.: Proc. of PACHEC’83, Vol. IV, p. 197 (The Third Pacific Chemical Engineering Congress) 1983
Wichmann, R., Wandrey, C., Bueckmann, A. F. and Kula, M.-R.: Biotech. Bioeng. 23, 2789 (1981)
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1989 Springer-Verlag Berlin Heidelberg
About this paper
Cite this paper
Kajiwara, S., Maeda, H. (1989). The Construction of a Bioreactor for the Production of an Optically Active Compound. In: Fiechter, A., Okada, H., Tanner, R.D. (eds) Bioproducts and Bioprocesses. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-74227-9_9
Download citation
DOI: https://doi.org/10.1007/978-3-642-74227-9_9
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-74229-3
Online ISBN: 978-3-642-74227-9
eBook Packages: Springer Book Archive