• Title/Summary/Keyword: cephalosporin-C

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Effect of Methionine on Cephalosporin C Production in a Fluidized- bed Bioreactor (유동층 생물반응기에서 세파로스포린 C 생산에 대한 메치오닌의 영향)

  • Kim, Eui-Yong;Yoo, Young-Je;Park, Young-Hoon
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.611-618
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    • 1989
  • Effects of methionine on cephalosporin C(CPC) production in a fluidized-bed bioreactor were investigated using bioparticles of Cephalosporium acremonium. Since methionine was found to be an important metabolic regulator on the synthesis of cephalosporin C, the effects of its concentration in the cuture broth and feeding mode to the bioreactor were studied. It was observed that the presence of initial methionine was essential for higher cephalosporin C production and there existed an optimal content of methionine. Carbon consumption rate also increased significantly under the presence of methionine. Production of cephalosporin C was most active when methionine was exhausted in the broth; however its additional feeding did not enhance the antibiotic production in the fluidized-bed bioreactor as much as expected. It was therfore considered important to feed an optimal content of methionine at the early operating stage for a higher cephalosporin C production in a fluidized-bed bioreactor. An interesting thing to note was that titre of the antibiotic with reused bioparticles was about 2 times higher in the methionine containing medium than that without methionine. Therefore repeated use of bioparticles, with an optimal content of methionine, was believed to be very useful to enhance to process productivity.

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Cloning and Expression of D-amino Acid Oxidise from Trigonopsis variabilis for Cephalosporin C Biotransformation (Cephalosporin C의 생변환을 위한 Trigonopsis variabilis의 D-amino Acid Oxidase 유전자의 클로닝 및 발현)

  • 이진형;정태완
    • KSBB Journal
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    • v.10 no.3
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    • pp.264-270
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    • 1995
  • Trigonopsis variabilis is a strong producer of D-amino acid oxidase that can transform cephalosporin C(ceph C) to ${\alpha}$-keto-adipyl-7-aminocephalosporanic acid(AKA-7ACA). Polymerase chain reaction (PCR) was applied to isolate the D-AAO gene from T. variabilis. To clone the PCR fragment, four different methods were examined using enzymatic reactions of Taq DNA polymerase, Klenow, T4 DNA polymerase I, Alkaline phosphatase Calf Intestinal, and T4 kinase. Ligation of phosphorylated blunt-end PCR fragment and dephosphorylated blunt-end of pUC18 plasmid yielded the best cloning efficiency One of recombinant E. coli transformants showed D-AAO activity against ceph C in both cell extracts and permeabilized cells.

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Isolation of Novel Pseudomonas diminuta KAC-1 Strain Producing Glutaryl 7-Aminocephalosporanic Acid Acylase

  • Kim, Dae-Weon;Kang, Sang-Mo;Yoon, Ki-Hong
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.200-205
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    • 1999
  • 7-Aminocephalosporanic acid (7-ACA) is the initial compound in preparation of cephalosporin antibiotics widely used in clinical treatment. Bacteria producing glutaryl 7-ACA acylase, which convert cephalosporin C to 7-ACA, has been screened in soil samples. A bacterial strain exhibiting high glutaryl 7-ACA acylase activity, designated KAC-1, was isolated and identified as a strain of Pseudomonas diminuta by characterizing its morphological and physiological properties. The screening procedures include culturing on enrichment media containing glutaric acid, glutamate, and glutaryl 7-aminocephalosporanic acid as selective carbon sources. To enhance enzyme production, optimal cultivation conditions were investigated. This strain grew optimally at pH 7 to 9 and in temperatures of 20 to 40 C, but acylase production was higher when the strain was grown at 25 C. Glutaric acid, glutamate and glucos also acted as inducers for acylase production. In a jar fermenter culture, P. diminuta KAC-1 produce acylase in a growth-associated manner. The substrate specificity of KAC-1 acylase by cell extract showed that this enzyme had specificity toward glutaryl 7-ACA, glutaryl 7-ADCA, but not cephalosporin C.

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Biosynthesis of $\beta$-Lactam Antibiotics by Cell-free Extract from Lysobacter lactamgenus

  • Roh, Ju-Won;Nam, Doo-Hyun
    • Archives of Pharmacal Research
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    • v.15 no.3
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    • pp.234-238
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    • 1992
  • Using cell-free extract of Lysobacter lactamgenus, enzymatic conversion of $\delta$-L-($\alpha$-aminoadiphyl)-L-cysteinyl-D-valine (ACV) the first substrate of $\beta$-lactam biosynthesis, into antibiotic compounds was attempted. In high performance liquid chromatographic (HPLC) analysis, the biosynthetic intermediates for cephalosporin antibiotics including isopenicillin N, deacetoxycephalosporin C, deacetylcephalosporin C and unknown cephem compound were detected in reaction mixtures. It implies that cephabacin compounds from L lactamgenus could be produced by biosynthetic routes through penicillin ring formation and its expansion to cephalosporin ring, likely as cephalosporin C from Cephalosporium or cephamycin C from Streptomyces. Among biosynthetic enzyme in cell-free extract, the ring formation activity (isopenicillin N synthetase activity) was separated in 50-60% of ammonium sulfate fraction, and ring expansion activity (deacetoxycephalosporin C synthetase activity) was found to be in 40-50% fraction. The partially purified isopenicillin N synthetase could convert as much as 90% ACV to isopenicillin N during 6-hour reaction.

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Effects of Nutrients and Culture Conditions on Morphology in the Seed Culture of Cephalosporium acremonium ATCC 20339

  • 이명선;김창호;오경권;홍석인;김승욱
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.2
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    • pp.156-156
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    • 2001
  • The objective of this study was to investigate the effects of nutrients and culture conditions on morphology during the seed culture of C. acremonium ATCC 20339 Morphological factors such as hyphal length number of tips number of arthrospores were observed to investigate the relationship between seed morphology and CPC production. During the time course of seed culture, hyphal length was shortened and the number of arthrospores increased rapidly On the other hand the number of tips deceased rapidly and this was closely related to the hyphal length Mixed nitrogen sources of 3% solybean meal and 1% cotton seed flour were determined as the proper organic nitrogen sources, in terms of the morphological factors in the seed culture. This fact was proven in batch culture for the production of Cephalosporin C. It was also found that a proper agitation speed enhanced the morphological differentiation of C. acremonium ATCC 20339, thus improving the production of Cephalosporin C.

Mechanism of Phosphate Regulation of Cephalosporin C Biosynthesis in Cephalosporium acremonium (Cephalosporium acremonium의 Cephalosporin C 생합성에 있어 무기인의 조절기작)

  • Choi, Sang-Ho;Lee, Kyoung;Yoon, Byung-Dae;Mheen, Tae-Ick
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.46-50
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    • 1989
  • A high concentration of inorganic phosphate (above 25 mM), which was suboptimal for vegetative growth in the minimal production medium, suppressed cephalosporin C (CPC) production in Cephalosporium acremonium. Results from the determination of intracellular concentrations of ATP, ADP and AMP with phosphate-starved resting cells indicated that phosphate exerted its effect indirectly by regulating the ratio of adenylated nucleotides, the so-called adenylated energy charge. It was also found that the type of phosphate regulation of CPC biosynthesis was not a repression effect but an inhibition effect.

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An Antibiotic from Actinomycetes Becoming Effective for Cephalosporin Resistant Pathogenic Pesudomonas sp. (방선균이 생산하는 Cephalosporin 내성 병원성 Pseudomonas에 유효한 항생물질)

  • 하병조
    • The Korean Journal of Food And Nutrition
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    • v.12 no.3
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    • pp.271-278
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    • 1999
  • We isolated activnmycetes LAM-98-80 as strain producing an effective antibiotic for cephalosporin re-sistant pathogenic PSeudomonas sp. and identified as Streptomyces sp. LAM-98-80 from cultural and phyisological characteristics. We investigated the optimal culture conditions for producation of an anti-biotic becoming effective for cephalsporin-resistant pathogenic Pseudomonas sp. It was found that 1.5% soluble starch and 1.0% yeast extract were good as carbon and nitrogen source respectively. The pro-duction of antibiotic was also activated by 0.04% Mn2+ as 80% degree. The optimum initial pH on pro-ductio of antibiotic was pH 7.0. The culture condition for the maximal productivity of the antibiotic was at 3$0^{\circ}C$ for 5 days. The cephalosporin-resistant pathogenic Pseudomonas sp. as test bacteria was rev-ealed to resist antibiotic of cepha families but revealed to not resist those of $\beta$-lactam families ampicil-lin and amoxicillin. Parital purified antibiotic was stable for the pH from 3 to 9 and was also stable when treated at 70 $^{\circ}C$ for 1 hour, This antbiotic was effective against all gram positive and negative bac-teria but was not effective against molds and yeasts.

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Synthesis and In Vitro Antibacterial Activity of C-3' Pyridinium Cephalosporin Derivatives

  • Chung, In-Hwa;Kim, Choong-Sup;Seo, Jae-Hong;Chung, Bong-Young
    • Archives of Pharmacal Research
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    • v.22 no.4
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    • pp.391-397
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    • 1999
  • The quaternary ammonium cephalosporin derivatives were prepared with various pyridines substituted at the 3 or/and 4 position. Their in vitro antibacterial activities were determined and substituent effect on pyridine nucleus was studied. Preparation of substituted pyridines are also described.

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Theoretical Studies on the Methanolysis of a Cephalosporin; Mimicking Acylation of the Active Site Serine of D-Ala-D-Ala Transpeptidases

  • Nahm, Kee-Pyung
    • Bulletin of the Korean Chemical Society
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    • v.12 no.6
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    • pp.674-678
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    • 1991
  • Methanolysis of a ${\beta}$-lactam ring of a cephalosporin was simulated with AM1 semiempirical quantum mechanical calculation. The tetrahedral intermediate TD1 from an O-protonated cephalosporin and a methanol transfers the proton intramolecularly to the C-4 carboxylate to generate an oxyanion, i.e., second tetrahedral intermediate TD2, which undergoes the amide bond cleavage without further protonation on the N-5. For this cleavage a low-energy barrier TS2 was located. According to the energy diagram, tetrahedral intermediates easily undergo ring cleavage even without the protonation on the amide nitrogen.

Studies of Repeated Fed-Batch Fermentation of Cephalosporin C in an Immobilized Cell Bioreactor

  • Park, Hong-Je;Khang, Yong-Ho
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.229-233
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    • 1995
  • Acremonium chrysogenum was immobilized in ionotropic gel beads to develop semi-continuous production of cephalosporin C (CPC). Barium alginate beads were more stable than calcium alginate or strontium alginate beads in chemically defined media. The gel stability of Ba-alginate was further increased by cross-linking with polyethyleneimine (PEI). The presence of carboxymethyl cellulose inside Ba-alginate beads did not reduce mass transfer resistance. Ba-alginate microbeads that had little diffusion limitation increased CPC production rate 1.6 fold higher than that of normal beads. CPC fermentation with immobilized cells in Ba-alginate microbeads was performed continuously for 40 days by way of repeated fed-batch operations. Mathematical modeling was developed to describe the repeated fed-batch fermentation system. Results of the computer simulation agreed well with the experimental data, which made it possible to predict an optimal feeding rate that could maximize total CPC productions.

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