As. Pac. J. Mol. Biol. & Biotech., June 2000 Vol 8(1) : 95-100
A colorimetric screening method for microorganisms having
methionine g-lyase activity
L.L. Choo 1, T. Tamura2, R.A. Rahim 1, A.M. Ali1, H. M. Ghazali1, K. Inagaki and H. Tanaka2*
1Department of Biotechnology, Faculty of Food Science and Biotechnology, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia
2Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University. 1 1 1,Abstract.
This paper reports a convenient screening method for detecting methionine g-lyase activity in microorganisms which can grow on DL methionine as the sole nitrogen source. L Methionine g-lyase has been known to have the most versatile catalytic properties among other pyridoxal 5' phosphate dependent enzymes, but only known from a limited number of biological sources. For an efficient screening of the enzyme, we employed 5, 5' dithiobis (2 nitrobenzoic acid) (DTNB) to detect the enzyme reaction product methanethiol, which spontaneously reduces DTNB and develops a yellow coloration on and around colonies. The yellow color has emerged and faded away within 72 h, suggesting that the reduced DTNB may be oxidized by spontaneous and/or enzymatic formation of disulfide. This method, despite reduction in specificity compared to the standard assay, allowed us to select candidate microorganisms out of hundreds of other similar colonies merely by observing the yellow coloration.
Key Words: L Methionine g-lyase, 5, 5' Dithiobis (2 nitrobenzoic acid)