• Microbiology and Biotechnology Letters
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• v.27 no.2
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• pp.96-101
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• 1999

Various side-chains are introduced to the 7-amino position of 7-aminocepha-losporanic acid (7-ACA) to make semi-synthetic cephalosporin antibiotics. In order to convert cephalosporin C (CPC) to 7-ACA, two enzymatic reactions are generally imployed. Glutary1-7-aminocephalosporanic acid (Gl-7-ACA) acylase is involved in the second step where the reaction intermediate, Gl-7-ACa is converted into 7-ACA. It was recently reported that CPC amidase can convert CPC directly into 7-ACA in a single enzymatic reaction. A study was undertaken to screen microorganisms conferring enzyme activity to convert Gl-7-ACA or CPC into 7-ACA by one or two enzymatic reactions. In order to screen the microorganisms rapidly, a non-$\beta$-lactam model compund, glutaryl-$\rho$-nitroanilide, was utilized in an early stage, thereafter the selected microorganisms were examined with real substrates. One microorganism exhibiting both Gl-7-ACA acylase and CPC amidase activities was obtained by the colorimetry method and HPLC assay, and was identified as a strain of Serratia species, designated as Serratia sp. N14.4. The optimal fermentation conditions for Serratia sp. N14.4 was pH9.0 and 3$0^{\circ}C$.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.6
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• pp.459-464
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• 2004

The objective of this study is to improve cephalosporin C (CPC) production byoptimization of medium and culture conditions. A statistical method was introduced to optimize the main culture medium. The main medium for CPC production was optimized using a statistical method. Glucose and corn steep liquor (CSL) were found to be the most effective factors for CPC production. Glucose and CSL were optimized to 2.84 and $6.68\%$, respectively. CPC produc­tion was improved $50\%$ by feeding of $5\%$ rice oil at day 3rd and 5th day during the shake flask culture of C acremonium M25. The effect of agitation speeds on CPC production in a 2.5-L bio­reactor was also investigated with fed-batch mode. The maximum cell mass (54.5 g/L) was obtained at 600 rpm. However, the maximum CPC production (0.98 g/L) was obtained at 500 rpm. At this condition, the maximum CPC production was improved about $132\%$ compared to the re­sult with batch flask culture.

• 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.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1120-1124
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• 2006

Supplementation with rice oil and its major components (oleic acid and linoleic acid) was found to have a significant influence on cephalosporin C (CPC) production and cell growth by A. chrysogenum M35 in shake flask cultures. Five percent (v/v) rice oil had the most robust effect and 5% (v/v) oleic acid was the second most efficient on cell growth, whereas 3% (v/v) linoleic acid was found to be optimal for CPC production. Rice oil, oleic acid, and linoleic acid also significantly improved the rates of glucose consumption. When glucose was almost consumed, CPC production was initiated and, on the addition of rice oil, lipase activity increased steadily to 1.56 U/ml for 4 days. These results suggest that rice oil and fatty acids are used as carbon source to produce CPC by A. chrysogenum M35. Moreover, a mixture, composed of 40% (v/v) oleic acid and 60% (v/v) linoleic acid, had the strongest stimulatory effect on CPC production, due to a synergistic effect of the two fatty acids. Consequently, the maximum CPC titer (7.44 g/l) was improved about 4.5-fold.

• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.296-303
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• 1994

An immobilized Trigonopsis variabilis cells having an high activity of D-amino acid oxidase(DAO) was used to convert CPC into GL-7-ACA. The optimal pH of the reaction system was 8.0-8.5, and the optimal temperature was 40$\circ$C. When immobilized cell was used repeatedly in semi-batchwise reaction, the system retained 80% of the initial activity after used of 12 times for over 12 hours. The storage stability of the immobilized cell was maintained for 30 days at 4$\circ$C. The CPC concentration for the maximal reaction rate was about 30 mM and 40 mM for free and immobilized cells, respectively. Substrate inhibition of CPC concentration more than 50 mM was overcomed by 20~25% by immobilization. Pure oxygen supply into reaction system was most efficient in D-amino acid oxidase reaction. Continuous conversion to GL-7-ACA from CPC has been developed with an bioreactor system containing immobilized T variabilis cells. By opera- tion of the reactor for 5 hours, the average conversion yield of >80% and GL-7-ACA production of 40~45 mM per hour could be obtained.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.339-344
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• 1985

Cephalosporin C(CPC) inhibited the growth of Cephalosporium acremonium M-113, a potent CPC producer derived from C acremonium ATCC 20339. Similar inhibitory effects of CPC were also observed in growth of C. acremonium ATCC 20339 and ATCC 14553. Minimum inhibitory concentrations (MIC) of CPC on the growth of conidia and hyphae of C. acremonium M-113 were 200-500 and 3000-4000$\mu\textrm{g}$/$m\ell$ respectively in synthetic medium. MIC values were increased in complex media. The inhibitory effect of CPC was due to CPC-exerted inhibition of amino acids uptake by the cells. 3'-Group of CPC might be important in its inhibitory action. In audition, CPC itself could be utilized by the cells as a nitrogen source under nitrogen limited condition.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.178-182
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• 1992

For an industrial-scale purification and production of cephalosporin C from a culture broth of Ceplzalos#mium nmemonium CSA-2.8A3 mutant, ultrafiltration, column chromatography, reverse osmosis, and spray drying were empolyed. Above 90% of yield and high purity of cephalosporin C were obtained through WA-30, HP-20, XAD-2000 and SK-1B column chromatographies. Especially, in the tendom operation of the columns, the recovery yield was increased up to 96%. The purified cephalosporin C was stable at $4^{\circ}C$ and in acidic condition, while it was unstable at room temperature and in alkaline condition at pH above 8.0. Cephalosporin C powder or a final product prepared by spray drying contained 85.554 of sodium cephalosporin C, 6.3%' of water, 4.63% of free $Na^+$ ions. and traces of metal ions.

• 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.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.224-229
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• 2004

Several microorganisms (esterase-producing group) were isolated by the solid selective media containing-naphtylacetate. Among them, strain 366M-9 having a high activity of cephalosporin-C deacetylase (CAH; EC 3.1.1.41) was selected. The strain 366M-9 was identified as Bacillus sphaericus on the basis of morphological, physiological, and biochemical characteristics. The production of CAH reached at maximum value after 32 hrs, when cultivated in the optimal medium containing dextrin 2.5%, peptone 2.5%, sodium chloride 0.5%, dipotassium phosphate 0.25%, ferrous sulfate 0.02%, and 7-ACA 0.1% at $30^{\circ}C$ with initial pH 6.0. The CAH was purified by 3 steps with ammonium sulfate precipitation, adsorption chromatography on hydroxyapatite column, and Sephadex G-200 gel chromatography. The final enzyme preparation was homogeneous as judged by the analysis of SDS-PAGE and HPLC. Optimum temperature and pH for CAH activity were $50{\circ}C$ and around 7.0, respectively. And the enzyme was stable at pH 6.0～8.0, up to $50^{\circ}C$. The Michaelis-Menten constants ($K_{m}$ ), $V_{max}$ were 0.87 mM and 1.22 unit/ml, respectively.