• Korean Journal of Microbiology
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• v.37 no.2
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• pp.164-169
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• 2001

The conversion of cephalosporin C to 7$\alpha$-hydroxycephalosporin C was examined with the cell-free extract of several cephamycin producing strains. Streptomyces clavuligerus ATCC 27064 was the most potent strain for the activity of cephalosporin 7$\alpha$-hydroxylase. Partially purified and immobilized cephalosporin 7$\alpha$-hydroxylase with resins were used to synthesize 7$\alpha$-hydroxycephalosporin C from the substrate, cephalosporin C. The molecular weight of the product isolated from the reaction mixture were determined to be 431 by ESI-Mass. $^1H$ NMR also support the conversion of cephalosporin C to 7$\alpha$-hydroxycephalosporin C by immobilized enzyme.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.417-426
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• 2008

The effect of increasing levels of proclavaminate amidino hydrolase (Pah) on the rate of clavulanic acid production in Streptomyces clavuligerus ATCC 27064 was evaluated by increasing dosoge of a gene (pah2) encoding Pah. A strain (SMF5703) harboring a multicopy plasmid containing the pah2 gene showed significantly retarded cell growth and reduced clavulanic acid production, possibly attributable to the deleterious effects of the multicopy plasmid. In contrast, a strain (SMF5704) carrying a single additional copy of pah2 introduced into chromosome via an integrative plasmid showed enhanced production of clavulanic acid and increased levels of pah2 transcripts. Analysis of transcripts of other genes involved in the clavulanic acid biosynthetic pathway revealed a pattern similar to that seen in the parent. From these results, it appears that clavulanic acid production can be enhanced by duplication of pah2 through integration of a second copy of the gene into chromosome. However, increasing the copy number of only one gene, such as pah2, does not affect the expression of other pathway genes, and so only modest improvements in clavulanic acid production can be expected. Flux controlled by Pah did increase when the copy number of pah2 was doubled, suggesting that under these growth conditions, Pah levels may be a limiting factor regulating the rate of clavulanic acid biosynthesis in S. clavuligerus.

• KSBB Journal
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• v.14 no.4
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• pp.440-444
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• 1999

For the effecient production of clavulanic acid., a mutant strain Streptomyces clavuligerus KK was selected from Streptomyces clavuligerus ATCC 27064 through mutation with NTG. S. clavuligerus ATCC 27064 produced about 200 mg/L of calvulanic acid when the medium was composed of 1%(W/V) glycerol, 1.5%(W/V) soybean flour, 0.1%(W/V) $KH_2PO_4$, 0.2%(V/V) soybean oil. A selected mutant, S. clavuligerus KK, produced about 1150 mg/L of clavulanic acid in the same medium. After the addition of $MgSO_4$ to the basal medium, S. clavuligerus KK produced about 1550 mg/L of clavulanic acid, with shows about 1.3 times higher than that produced in the basal medium. In order to select the proper bioreactor for the production of clavulanic acid, a batch culture was performed in an airlift, a bubble column and an stirred tank bioreactors. In an airlift bioreactor, about 1350 mg/L of clavulanic acid was produced, in a bubble column bioreactor, about 1550 mg/L, in a stirred tank bioreactor, about 2200 mg/L, respectively. The production of clavulanic acid in stirred tank bioreactor was about 50% higer than that by an airlift and a bubble column bioreactors. According to this result, the stirred tank bioreactor was selected as a proper bioreactor.