Application of lat gene disruption to increase the clavulanic acid production of Streptomyces clavuligerus
| Autor: | Wang Yan-ping, Zuo Zhihan |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
biology
Chemistry Process Chemistry and Technology Mutant Chromosome Streptomyces clavuligerus Bioengineering medicine.disease_cause biology.organism_classification Biochemistry Genome Molecular biology Catalysis Microbiology law.invention law Clavulanic acid medicine Recombinant DNA Escherichia coli Gene medicine.drug |
| Zdroj: | Journal of Molecular Catalysis B: Enzymatic. 43:102-107 |
| ISSN: | 1381-1177 |
| DOI: | 10.1016/j.molcatb.2006.06.018 |
| Popis: | A 1.7 kb fragment of lat was obtained from Streptomyces clavuligerus NRRL 3585, and recombinant plasmid pKC1139-lat, which was used to disrupt the lat gene was constructed. pKC1139-lat was introduced into S. clavuligerus by bi-parental conjugation from Escherichia coli ET12567 to S. clavuligerus. The apramcin-resistant transformants were obtained and through homogeneous single-crossover between recombinant plasmid pKC1139-lat and the S. clavuligerus chromosome lat disrupted mutant strains were obtained. The genome of S. clavuligerus NRRL 3585 and the lat disrupted mutants were analyzed by PCR technique, the bioactivity of cephamycin C in the two kinds of strains were also tested. Both results proved that lat was disrupted by the insertion of pKC1139 in the lat disrupted mutants. And the production of clavulanic acid of these two kinds of strains were analyzed by HPLC with different incubation time interval (96 and 120 h), and the yield in the lat mutants was approximately 2.6 fold higher at their highest production point. |
| Databáze: | OpenAIRE |
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