Integration of clavaminate synthase 2 gene into the chromosome of an industrial strain of Streptomyces Clavuligerus for enhanced clavulanic acid production

Download
index.pdf
2010
Vanlı, Güliz
Show full item record
491
views
91
downloads
Streptomyces clavuligerus is a gram-positive, filamentous bacterium which has a great ability to produce secondary metabolites including isopenicillin N, cephamycin C and a beta-lactamase inhibitor clavulanic acid. Clavulanic acid (CA) which is a bicyclic beta-lactam, inhibits most of class A beta-lactamases by binding irreversibly to the serine hydroxyl group at the active center of beta-lactamases and resulting in the stable acyl-enzyme complexes. Clavaminate synthase (CAS) is one of the best characterized enzymes in the CA biosynthesis pathway regarding its mechanism, activity and structure. It exists as two isoenzymes, CAS1 and CAS2. cas1 gene is located in clavam biosynthetic gene cluster and its expression is nutritionally regulated. cas2 gene encodes for the rate limiting CAS2 isoenzyme which catalyzes the three distinct oxidative transformations. Conversion of deoxyguanidinoproclavaminic acid into guadinoproclavaminic acid takes part in early steps of CA biosynthesis and is catalysed by CAS2. Similarly, proclaviminic acid is converted into claviminic acid by a two-step reaction and also catalysed by CAS2. Integration of an extra copy of the cas2 gene into the chromosome of a clavulanic acid over-producer industrial strain of Streptomyces clavuligerus by means of an integration vector to improve CA production capacity of the industrial strain is the main goal of this study. Via comperative CA analysis based on HPLC, it was estimated that the recombinant strains produced higher amount of CA in comparison to the parental industrial S. clavuligerus strain. The highest CA production achieved by the recombinant strain, namely S. clavuligerus GV61 (1583.3 μg/g) corresponded to more than 2 fold increase in the maximal CA titer of the parental strain.
Microbial genetic engineering., Genetic engineering, Genetic engineering., Biotechnology.
http://etd.lib.metu.edu.tr/upload/12612576/index.pdf
https://hdl.handle.net/11511/20098
Graduate School of Natural and Applied Sciences, Thesis

Suggestions

The regulatory effect of CcaR activator on the cephamycin C gene cluster of streptomyces clavuligerus
Kurt, Aslıhan; Özcengiz, Gülay; Department of Biology (2011)
Streptomyces clavuligerus produces industrially important secondary metabolites such as cephamycin C (a β-lactam antibiotic) and clavulanic acid (a potent β-lactamase inhibitor). Cephamycin C is active against penicillin-resistant bacteria due to presence of methoxyl group in C-7 position of cephalosporin nucleus. Clavulanic acid is prescribed in combination with β-lactams for treatment of various bacterial infections. Cephamycin C and clavulanic acid gene clusters form β-lactam supercluster in S. clavulige...
Genetic engineering of glycolytic pathway by disrupting glyceraldehyde-3-phosphate dehydrogenase gene in an industrial strain of streptomyces clavuligerus
Sertdemir, İbrahim; Özcengiz, Gülay; Department of Biology (2013)
Streptomyces clavuligerus is a gram-positive, filamentous bacterium which produces several important secondary metabolites, including isopenicillin N, cephamycin C and the β-lactamase inhibitor, clavulanic acid (CA). CA is being used in combination with commonly used β-lactam antibiotics in order to fight against bacterial infections that are resistant to such antibiotics. Glyceraldehyde-3-phosphate (GAP) which is an intermediate product of glycolytic pathway is also used in CA biosynthesis as a crucial sub...
Conducting graft copolymers of pyrrole and thiophene with random copolymers of methyl methacrylate and 3-methylthienyl methacrylate
Guner, Y; Toppare, Levent Kamil; Hepuzer, Y; Yagci, Y (2004-08-01)
Random copolymers of 3-methyl thienylmethacrylate and methyl methacrylate were synthesized via free radical polymerization. Electro-copolymerizations of random copolymers with thiophene and/or pyrrole were carried out in acetonitrile-tetrabutylammonium tetrafluoroborate (TBAFB), water-p-toluene sulfonic acid (PTSA) solvent-electrolyte couples. Oxidative polymerization of thiophene functionalized random copolymer was also achieved by constant current electrolysis and chemical polymerization. The characteriza...
Synthesis and characterization of thiophen-3-yl-acetic acid 4-pyrrol-1-yl-phenyl ester and its conducting polymers
Bingöl, Bahar; Toppare, Levent Kamil; Department of Chemistry (2003)
Thiophen-3-yl acetic acid 4-pyrrol-1-yl phenyl ester (TAPE) monomer was synthesized by the reaction of thiophene acetic acid with thionyl chloride, and further reaction of thiophen-3-yl-acetyl chloride with 4-pyrrol-1-yl phenol. Electrochemical behavior of this monomer (TAPE) was determined by cyclic voltammetry. Homopolymers were achieved both by using electrochemical and chemical polymerization techniques. Copolymers of TAPE in the presence of bithiophene and pyrrole were synthesized by potentiostatic ele...
Increasing clavulanic acid production both in wild type and industrial streptomyces clavuligerus strains by amplification of positive regulator claR gene
Mutlu, Alper; Özcengiz, Gülay; Department of Biology (2012)
Streptomyces clavuligerus is a Gram-positive, filamentous bacterium which produces several important secondary metabolites, including isopenicillin N, cephamycin C and the β-lactamase inhibitor clavulanic acid. Among these compounds, clavulanic acid is being used in combination with commonly used β-lactam antibiotics in order to fight against bacterial infections that are resistant to such antibiotics. Among these combinations, Augmentin, composed of amoxicillin and clavulanic acid, is the most widely presc...
Citation Formats
G. Vanlı, “Integration of clavaminate synthase 2 gene into the chromosome of an industrial strain of Streptomyces Clavuligerus for enhanced clavulanic acid production,” M.S. - Master of Science, Middle East Technical University, 2010.
METU IIS - Integrated Information Service open@metu.edu.tr