This paper reports a convenient screening method for detecting methionine gamma -lyase activity in microorganisms which can grow on DL-methionine as the sole nitrogen source. L-Methionine gamma -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.
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Royal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, AustraliaRoyal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, Australia
van Werkhoven, Michiel A.
Duley, John A.
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Univ Queensland, Sch Pharm, Brisbane, Qld, Australia
Mater Med Res Inst, Brisbane, Qld, AustraliaRoyal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, Australia
Duley, John A.
McGown, Ivan
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Mater Hlth Serv, Brisbane, Qld, AustraliaRoyal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, Australia
McGown, Ivan
Munce, Teresa
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Mater Hlth Serv, Brisbane, Qld, AustraliaRoyal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, Australia
Munce, Teresa
Freeman, Jeremy L.
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Royal Childrens Hosp, Dept Neurol, Melbourne, Vic 3052, Australia
Royal Childrens Hosp, Murdoch Childrens Res Inst, Melbourne, Vic 3052, AustraliaRoyal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, Australia
Freeman, Jeremy L.
Pitt, James J.
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Royal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, AustraliaRoyal Childrens Hosp, Murdoch Childrens Res Inst, Victorian Clin Genet Serv, Melbourne, Vic 3052, Australia
Pitt, James J.
DEVELOPMENTAL MEDICINE AND CHILD NEUROLOGY,
2013,
55
(11):
: 1060
-
1064