Biopolym. Cell. 1998; 14(2):127-131.
Structure and Function of Biopolymers
Phagedependent overproduction of β-galactosidase Escherichia coli and working out its purification
1Chernykh S. I., 1Slavchenko I. Yu., 1Gorlov Yu. I., 1Terent'ev A. G., 1Gavrish T. G., 1Kordyum V. A.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680


The phagedependent method of obtaining of β-galactosidase (EC E. coli has been worked out. The yield was up 700 U of enzyme activity per ml of cultural fluid. Accumulation of enzyme in cultural medium has permitted to use membrane method for preliminary purification of β-galactosidase. Concentrated and partially purified preparation of β-galactosidase has been purified by ligand-affinity chromatography. Pure preparation of β-galactosidase has characterized by high purity and nativeness. The process of obtaining and purification of β-galactosidase were made in pilot scale.


[1] Kaul R, D'Souza SF, Nadkarni GB. Hydrolysis of milk lactose by immobilized beta-galactosidase-hen egg white powder. Biotechnol Bioeng. 1984;26(8):901-4.
[2] Houts Sandra S. Lactose intolerance. Food Technol. 1988. 42(3):110-3.
[3] Gibbons I, Skold C, Rowley GL, Ullman EF. Homogeneous enzyme immunoassay for proteins employing beta-galactosidase. Anal Biochem. 1980;102(1):167-70.
[4] Freytag JW. Affinity column mediated immunoenzymometric assay. In: Enzyme-mediated immunoassay. Eds. TT Ngo, HM Lemhoff. Plenum Press New York 1985 277-298.
[5] Li XL, Robbins JW Jr, Taylor KB. The production of recombinant beta-galactosidase in Escherichia coli in yeast extract enriched medium. J Ind Microbiol. 1990;5(2-3):85-93.
[6] Kane JF, Hartley DL. Formation of recombinant protein inclusion bodies in Escherichia coli. Trends Biotechnol. 1988;6(5):95–101.
[7] AS. USSR N 1593232 Al. A method of producing ?-galactosidase. VA Kordyum, SI Chernykh, IYu Slavchenko, VG Korobko. 15.05.1990.
[8] Griffith KL, Wolf RE Jr. Measuring beta-galactosidase activity in bacteria: cell growth, permeabilization, and enzyme assays in 96-well arrays. Biochem Biophys Res Commun. 2002;290(1):397-402. Erratum in: Biochem Biophys Res Commun 2002 Mar 22;292(1):292.
[9] Affinity chromatography: a practical approach : Ed PD Dean, WS Johnson, FA Middle, IRL Press, Oxford, Washington, 1985, 215 p.