• 약학회지
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• 제30권6호
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• pp.294-300
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• 1986

New antibiotics having moieties of penicillanic acid, cephalosporanic acid and ampicillin on both ends of the central polyalkylene were synthesized by reacting 6-aminopenicillanic acid (6-APAl), 7-amino cephalosporanic acid (7-ACA) and ampicillin with hexamethylene diisocyanate and sebacoyl chloride, respecetively. Antibiacterial activities of the compounds were also investigated.

• 한국미생물·생명공학회지
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• 제22권3호
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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.

• 한국응용과학기술학회지
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• 제6권1호
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• pp.27-30
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• 1989

The fast separation and determination of cefatrizine${\cdot}$propylene glycol and inmpurities - TACA: 7-amino-3-(1,2,3-triazol-4-yl)thiomethyl-3-cephem-4-carboxylic acid and 7-ACA; 7-amino cephalosporanic acid - was performed by the high poerformance liquid chromatography using octadecyl siland (ODS) column. Methanol and ammonium phosphate buffer [$0.03M(NH_4)_2\;HPO_4$, (pH 7.5)] was used analyze, as eluent. The experimental value of the contents of cefatrizine${\cdot}$propylene glycol and impurities agree with the theoretical value of those.

• 한국미생물·생명공학회지
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• 제27권2호
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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$0^{\circ}C$.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1978년도 추계학술대회
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• pp.206.4-206
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• 1978

By utilizillg whole cell enzyme of the Xantho-monas citri IFO 3835, cephalexin is synthesized directly from 7-amino-deacetoxy cephalosporanic acid (7-ADCA) and phenyl glycine methyl ester (PGM). To date, cephalexin has been manufactu-red by chemical process involving fairly large number of steps to protect the amino group of phenly glycine and carboxyl group of 7-ADCA. However, the enzymatic process involves only a single step with 85％ conversion in 90 minutes. The fermentation variables studied indicate that oxygen transfer is limiting step in the enzyme production. Optimum conditions for enzymatic reaction were 37 C, pH 6.0, and the optimum substrate molar ratio of PGM to 7-ADCA was 2. Other variables that are related to the biochemical properties of whole cell enzyme temperature stability, pH stability, kinetic constants, reusing effect, enzyme loading effect were also evaluated.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.452-457
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• 2001

The glutaryl 7-aminocephalosporanic acid (glutaryl 7-ACA) acylase was purified from Pseudomonas diminuta KAC-1 cells isolated from soil, and characterized. The acylase was purified by procedures including ammonium sulfate fractionation and column chromatographies on DEAE-Sepharose, Phenyl-Sepharose, Q-Sepharose, and Superose 12H/R. The negative acylase was found to be composed of two subunits with molecular masses of approximately 55 kDa and 17 kDa, respectively. The isoelectric point of the enzyme was 4.0. The specific activities of the purified acylase were 8.0 and 7.0 U/mg on glutaryl 7-ACA and glutaryl 7-aminodesacetoxy cephalosporanic acid (glutaryl 7-ADCA), respectively, and $K_m$ values were 0.45 mM for glutaryl 7-ADCA and 0.67 mM for glutaryl 7-ADCA. The enzyme had a pH optimum at 8.0 and a tmperature optimum at $40^{\circ}C$. The acylase catalyzed the synthesis of glutaryl 7-ACA from glutaric acid and 7-ACA as well as the hydrolysis of glutaryl 7-ADCA, although the reaction rate of the synthesis was slower than that of the hydrolysis. In addition, it was found that the enzyme had a glutaryl transferase activity, thereby transferring the glutaryl group from one cephalosporin nucleus to another.

• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.155-162
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• 2020

Acetyl xylan esterase (AXE; E.C. 3.1.1.72) is one of the accessory enzymes for xylan degradation, which can remove the terminal acetate residues from xylan polymers. In this study, two genes encoding putative AXEs (LaAXE and BhAXE) were cloned from Lactobacillus antri DSM 16041 and Bacillus halodurans C-125, and constitutively expressed in Escherichia coli. They possess considerable activities towards various substrates such as p-nitrophenyl acetate, 4-methylumbelliferyl acetate, glucose pentaacetate, and 7-amino cephalosporanic acid. LaAXE and BhAXE showed the highest activities at pH 7.0 and 8.0 at 50℃, respectively. These enzymes are AXE members of carbohydrate esterase (CE) family 7 with the cephalosporine-C deacetylase activity for the production of antibiotics precursors. The simultaneous treatment of LaAXE with Thermotoga neapolitana β-xylanase showed 1.44-fold higher synergistic degradation of beechwood xylan than the single treatment of xylanase, whereas BhAXE showed no significant synergism. It was suggested that LaAXE can deacetylate beechwood xylan and enhance the successive accessibility of xylanase towards the resulting substrates. The novel LaAXE originated from a lactic acid bacterium will be utilized for the enzymatic production of D-xylose and xylooligosaccharides.