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Thermostable malate synthase of Streptomyces thermovulgaris.
- Source :
-
Journal of industrial microbiology & biotechnology [J Ind Microbiol Biotechnol] 2003 Oct; Vol. 30 (10), pp. 577-81. Date of Electronic Publication: 2003 Sep 06. - Publication Year :
- 2003
-
Abstract
- The gene, encoding malate synthase (MS), aceB, was cloned from the thermophilic bacterium Streptomyces thermovulgaris by homology-based PCR. The 1,626-bp cloned fragment encodes a protein consisting of 541 amino acids. S. thermovulgaris malate synthase (stMS) gene was over-expressed in Escherichia coli using a glutathione-S transferase (GST) fusion vector (pGEX-6P-1), purified by affinity chromatography, and subsequently cleaved from its GST fusion partner. The purified stMS was characterized and compared to a mesophilic malate synthase (scMS) from Streptomyces coelicolor. stMS exhibited higher temperature optima (40-60 degrees C) than those of scMS (28-37 degrees C). It was more thermostable and very resistant to the chemical denaturant urea. Amino acid sequence comparison of stMS with four mesophilic streptomycete MSs indicated that they share 70.9-91.4% amino acid identities, with stMS possessing slightly more charged residues (approximately 31%) than its mesophilic counterparts (approximately 28-29%). Seven charged residues (E85, R187, R209, H239, H364, R382 and K520) that were unique to stMS may be selectively and strategically placed to support its peculiar characteristics.
- Subjects :
- Amino Acid Sequence
Cloning, Molecular
Enzyme Activation
Gene Expression Regulation, Bacterial
Gene Expression Regulation, Enzymologic
Malate Synthase isolation & purification
Malate Synthase metabolism
Protein Denaturation
Hot Temperature
Industrial Microbiology methods
Malate Synthase genetics
Streptomyces enzymology
Streptomyces genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1367-5435
- Volume :
- 30
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Journal of industrial microbiology & biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 13680388
- Full Text :
- https://doi.org/10.1007/s10295-003-0082-9