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Enzymatic synthesis of γ-L-glutamyl-S-allyl-L-cysteine, a naturally occurring organosulfur compound from garlic, by Bacillus licheniformis γ-glutamyltranspeptidase.
- Source :
-
Enzyme and microbial technology [Enzyme Microb Technol] 2015 Jul-Aug; Vol. 75-76, pp. 18-24. Date of Electronic Publication: 2015 May 04. - Publication Year :
- 2015
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Abstract
- In the practical application of Bacillus licheniformis γ-glutamyltranspeptidase (BlGGT), we describe a straightforward enzymatic synthesis of γ-L-glutamyl-S-allyl-L-cysteine (GSAC), a naturally occurring organosulfur compound found in garlic, based on a transpeptidation reaction involving glutamine as the γ-glutamyl donor and S-allyl-L-cysteine as the acceptor. With the help of thin layer chromatography technique and computer-assisted image analysis, we performed the quantitative determination of GSAC. The optimum conditions for a biocatalyzed synthesis of GSAC were 200 mM glutamine, 200 mM S-allyl-L-cysteine, 50 mM Tris-HCl buffer (pH 9.0), and BlGGT at a final concentration of 1.0 U/mL. After a 15-h incubation of the reaction mixture at 60 °C, the GSAC yield for the free and immobilized enzymes was 19.3% and 18.3%, respectively. The enzymatic synthesis of GSAC was repeated under optimal conditions at 1-mmol preparative level. The reaction products together with the commercially available GSAC were further subjected to an ESI-MS/MS analysis. A significant signal with m/z of 291.1 and the protonated fragments at m/z of 73.0, 130.1, 145.0, and 162.1 were observed in the positive ESI-MS/MS spectrum, which is consistent with those of the standard compound. These results confirm the successful synthesis of GSAC from glutamine and S-allyl-L-cysteine by BlGGT.<br /> (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Subjects :
- Bacterial Proteins metabolism
Cysteine analogs & derivatives
Cysteine metabolism
Dipeptides isolation & purification
Enzymes, Immobilized metabolism
Glutamine metabolism
Hydrogen-Ion Concentration
Industrial Microbiology
Recombinant Proteins metabolism
Spectrometry, Mass, Electrospray Ionization
Sulfur Compounds metabolism
Tandem Mass Spectrometry
Temperature
Bacillus enzymology
Dipeptides biosynthesis
Garlic metabolism
gamma-Glutamyltransferase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1879-0909
- Volume :
- 75-76
- Database :
- MEDLINE
- Journal :
- Enzyme and microbial technology
- Publication Type :
- Academic Journal
- Accession number :
- 26047911
- Full Text :
- https://doi.org/10.1016/j.enzmictec.2015.04.011