Enzymes Beta-Galactosidase

Ljungcrantz, P., Carlsson, H., Mansson, M. O., Buckel, P., Mosbach, K., and Bulow, L. (1989). Construction of an artificial bifunctional enzyme, beta-galactosidase/galac-tose dehydrogenase, exhibiting efficient galactose channeling. Biochemistry, 28,8786— 8792. 

Enzyme labels are currently by far the most common labels in use. Enzymes include HRP, alkaline phosphatase (AP) and, less often, beta-galactosidase (P-gal). The effectiveness of the conjugated antibody depends on the antibody, the label, and the procedure chosen to link the two.30... 

In in vitro studies penicillamine inhibited angiotensin-con-verting enzyme (ACE) and carboxypeptidase (930). Penicillamine interferes with the functions of the copper-containing enzyme ceruloplasmin, and some of the penicillamine- and copper-containing complexes formed in vivo have a superoxide dismutase effect (931). In patients with scleroderma, penicillamine normalized collagen metabolism, by inhibiting beta-galactosidase activity (932). 

Rossi F, Charlton CA, Blau HM. Monitoring protein-protein interactions in intact eukaryotic cells by beta-galactosidase complementation. Proc. Natl. Acad. Sci. U.S.A. 1997 94 8405-8410. Wehrman TS, Casipit CL, Gewertz NM, Blau HM. Enzymatic detection of protein translocation. Nat. Methods 2005 2 521-527. Hammer MM, Wehrman TS, Blau HM. A novel enzyme complementation-based assay for monitoring G-protein-coupled receptor internalization. FASEB J. 2007 21 3827-3834. 

Enzyme catalyzed reactions have also been studied at the single molecule level. Earlier work involved measurement of beta-galactosidase activity in droplets after a 10-15 h incubation. In a recent study, detection of fluorescent product generated by individual molecules of lactate dehydrogenase after a 1 hr incubation has been achieved using capillary electrophoresis . The activity of individual molecules were reproducible but activity of different molecules showed a 5-fold range. The differences in activity were suggested to reflect differences in conformation. 

S. Lee and B. H. Kim, frans-Sialidase catalyzed sialylation of beta-galactosyldisaccharide with an introduction of beta-galactosidase, Enzyme Microb. Technol., 28 (2001) 161-167. 

Tominaga, L., Ogawa, Y., Taniguchi, M., Ohno, K., Matsuda, J., Oshima, A., Suzuki, Y., and Nanba, E., Galactonojirimycin derivatives restore mutant human beta-galactosidase activities expressed in fibroblasts from enzyme-deficient knockout mouse, Brain Develop 23 (2001) 284 287. 

Baur X, Sander I, Jansen A et al. (1994) Are amylases in bakery products and flour potential food aUeigens Schweiz Med Wochenschr 124(20) 846-851 Becerra M, Cerdan E, Gonzalez Siso MI (1997) Heterologous Kluyveromyces lactis beta-galactosidase production and release by Saccharomyces cerevisiae osmotic-remedial thermosensitive autolytic mutants. Biochim Biophys Acta 1335(3) 235-241 Bertoldo C, Antranikian G (2002) Starch-hydrolyzing enzymes from thermophilic archaea and bacteria. Curr Opin Chem Biol 6 151-160... 

Decleire M, De Cat W, van Huynh N (1987) Comparison of various permeabilization treatments on Kluyvewmyces by determining in situ beta-galactosidase activity. Enzyme Microb Technol 9(5) 300-302... 

Competitive enzyme-linked immunosorbent assays (ELISA) Two types of ELISA have been used for the analysis of mycotoxins and both types are heterogenous competitive assays. One type, i.e. direct ELISA, involves the use of a mycotoxin-enzyme conjugate and the other system, i.e. indirect ELISA, involves the use of a protein-mycotoxin conjugate and a secondary antibody to which an enzyme has been conjugated. Although horseradish peroxidase (HRP) is most commonly used as the enzyme for conjugation, other enzymes such as alkaline phosphatase and beta-galactosidase, also have been used (5, 9, 13). 

H. Morreau, N. J. Galjait, N. Gillenmns, R. Willemsen, G. T. van der Horst, and A. d Azzo, Alternative sphcing of beta-galactosidase mRNA generates the classic lysosomal enzyme and a beta-galactosidase-related protein, J. Biol. Chem., 264 (1989) 20655-20663. 

Because of the need for sensitivity, most labels for DNA analysis are enzymes or highly efficient fluor-ophores such as phycobiliproteins. The most commonly used enzyme labels are calf intestinal alkaline phosphatase and horseradish peroxidase (Table 2). Beta-galactosidase is used primarily for gene expression monitoring. Other enzymes such as glucose-6-phosphate dehydrogenase have not fovmd as wide utility as phosphatase and peroxidase. 

Ansari, S. A. Satar, R. Recombinant beta-galactosidases— past, present and future a mini review. J. Mol. Catal. B Enzym. 2012, 81, 1-6. 

Kaftzik, N., Wassercheid, P., and Kragl, U., Use of ionic liquids to increase the yield and enzyme stability in the beta-galactosidase catalyzed synthesis of N-acetylactosamine, Org. Proc. Res. Dev., 6 553-557, 2002. 

Stred anskJ, M., M. Tomaska, E. Sturdik, and L. Kremnicky. 1993. Optimization of beta-Galactosidase Extraction from Kluyveromyces Marxianus. Enzyme and Microbial Technology 15 (12) 1063-1065. 

Kamerke, C., Pattky, M., Huhn, C., and filling, L (2012) Synthesis of UDP-activated oligosaccharides with commercial beta-galactosidase from Bacillus circulans under microwave irradiation. J. Mol. Catal. B Enzym., 79, 27-34. 

Rodriguez-Colinas B, de Abreu MA, Femandez-Arrojo L, de Beer R, Poveda A,Jimenez-Barbero J, et al. Production of galacto-ohgosaccharides by the beta-galactosidase from Kluyveromyces lactis comparative analysis of permeabUized cells versus soluble enzyme.] Agric Food Chem 2011 59(19) 10477-84. 

Walsh R., Martin E. Darvesh S. (2007). A versatile equation to describe reversible enzyme inhibition and activation kinetics modeling beta-galactosidase and butyrylcholinesterase. Biochim. Biophys. Acta, 1770, 733-746. 

Burin, L. Buera, M. P. (2002). Beta-galactosidase activity as affected by apparent pH and physical properties of reduced moisture systems. Enzyme andMicr. Techn., 30, 367-373. 

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