• 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.23 no.5
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• pp.559-564
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• 1995

Twenty microbial strains producing the acylase were isolated from soil by using Micrococcus luteus ATCC 9341 as an indicator strain, using either D-($\alpha $)-phenylglycine methylester and 7-aminocephalosporanic acid (7-ACA) or glutaric acid dimethylester and 7-ACA as substrates. Among the isolates, only one strain was turned out to be the 7-ACA producer from either cephalosporin C or glutaryl 7-ACA as the substrates by using the overlay of 7-ACA sensitive strain (SS5). 7-ACA produced from cephalosporin C by an isolate (APS20) was detected by high performance liquid chromatography. The isolated strain (APS20) was identified to Bacillus macerans on the basis of cellular fatty acid profile by gas chromatography. Bacillus macerans APS20 had no $\beta $-lacta-mase activity on cephalosporin C, and that is very important for the enzymatic production process of 7-ACA. However, this strain was resistant up to 100 $\mu $g/ml of cephalosporin C.

• Journal of Microbiology and Biotechnology
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• v.6 no.5
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• pp.336-339
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• 1996

Glutaryl 7-aminocephalosporanic acid acylase of Pseudomonas sp. SY-77-1 was immobilized with oxiran acrylic beads for the production of 7-aminocephalosporanic acid (7-ACA) from glutaryl 7-aminocephalosporanic acid (GL 7-ACA). The immobilized enzyme maintained its activity at a constant level for 7 days, but lost 30$%$ of its activity after 20 days. Optimal reaction conditions for the synthesis of 7-ACA were found to be $30^{\circ}C$ and pH 8.0 using the immobilized enzyme. For the economic production of 7-ACA, substrate and enzyme concentrations were optimized to 60 mM and 0.5 g wet weight per 10 $m\ell$ of reaction volume, respectively. Under optimized conditions, 50 mM 7-ACA was produced from 60mM GL 7-ACA within 8 h, resulting in a conversion yield of 83$%$.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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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
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.105-108
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• 1997

A search was undertaken to screen microorganisms that produce an enzyme capable of deacylating glutary1-7-amincephalosporanic acid to 7-aminocephalosporanic acid in soil samples. The screening was carried out by preparing enrichment cultures containing glutary-7ACA and cephalosporin C as selective carbon sources. A non-${\beta}$-lactam model compound,, glutary-p-nitroanilide, was synthesized as a substrate suitable for the rapid screening of microorganisms isolated from the enrichment cultures. Two isolates exhibiting acylase activity, designated BY7.4 and BY8.1, were identified as strains of Pseudomonas species. Pseudomonas BY8.1 showed higher acylase activity toward G1-7ACA than Pseudomonas BY7.4. Environmental conditions for the optimal acylase activity of Pseudomonas BY8.1 were shown to be pH9 and 30$^{\circ}C$.

• Korean Journal of Materials Research
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• v.5 no.8
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• pp.929-935
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• 1995

In order to develop COG (Chip On Glass) technology for LCD module interconnecting the driver IC to Al pad electrode on the glass substrate, Anisotropic Conductive Adhesive(ACA) process, the most promising one among COG technologies, was investigated. ACA process was carried out by two steps, dispensing of ACA resin in the bonding area and curing by W radiation. Load on the chip was ranged from 2.0 to 15kg and the chip was heated at about 12$0^{\circ}C$. In resin, the density of conductive particles coated with Au or Ni at the surface were 500, 1000, 2000 and 4000 particles/$\textrm{mm}^2$, and the diameter of particles were 5, 7 and 12${\mu}{\textrm}{m}$. As a result of the experiments, ACA process using ACA particle of diameter and density of 5${\mu}{\textrm}{m}$ and 4000 particles/$\textrm{mm}^2$ respectively shows optimum characteristic with the stabilzed bonding properties and contact resistance.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.04a
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• pp.121.2-122
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• 2003

다슬기분말(PSB)과 그 회분(ASB)으로 제조한 칼슘락테이트(PCaL 및 ACaL)를 0.5%씩 첨가한 반죽의 발효와 빵의 품질 및 저장성에 미치는 영향을 조사하였다. 반죽의 pH는 4.85~4.98로 ACaL.PCaL.대조군의 순으로 나타났다. 반죽의 부피와 빵의 loaf volume index는 대조군이 높았고 ACaL 첨가군이 낮았으며, pH를 5.50으로 조정하여 제조한 반죽의 부피와 빵의 loaf volume은 대조군과 큰 차이를 보이지 않았다. PCaL 및 ACaL을 첨가한 빵의 Ca함량은 29.4~29.7 mg/100 g-f.w로 대조군의 13.0 mg/100 g-f.w에 비하여 높았으며, 첨가군의 미량 무기질로 Mg, Fe, Zn이 0.03~0.98 mg/100 g-f.w 범위로 검출되었다. 빵의 L$^{*}$ 값은 대조군과 실험군의 유의적인 차이가 없었으며, a＊값, b＊값은 PCaL 첨가군이 가장 높아 황갈색을 띄었다. 빵의 hardness, gumminess는 대조군$^{\circ}C$ 실온에 두면서 저장한 결과 대조군은 3일째부터, ACaL 첨가군은 5일째부터, PCaL 첨가군은 6일째부터 곰팡이가 번식하였다.

• 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.

• Journal of the Microelectronics and Packaging Society
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• v.12 no.1 s.34
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• pp.9-16
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• 2005

This paper presents the development of new anisotropic conductive adhesives with enhanced thermal conductivity for the wide use of adhesive flip chip technology with improved reliability under high current density condition. The continuing downscaling of structural profiles and increase in inter-connection density in flip chip packaging using ACAs has given rise to reliability problem under high current density. In detail, as the bump size is reduced, the current density through bump is also increased. This increased current density also causes new failure mechanism such as interface degradation due to inter-metallic compound formation and adhesive swelling due to high current stressing, especially in high current density interconnection, in which high junction temperature enhances such failure mechanism. Therefore, it is necessary for the ACA to become thermal transfer medium to improve the lifetime of ACA flip chip joint under high current stressing condition. We developed thermally conductive ACA of 0.63 W/m$\cdot$K thermal conductivity using the formulation incorporating $5 {\mu}m$ Ni and $0.2{\mu}m$ SiC-filled epoxy-bated binder system to achieve acceptable viscosity, curing property, and other thermo-mechanical properties such as low CTE and high modulus. The current carrying capability of ACA flip chip joints was improved up to 6.7 A by use of thermally conductive ACA compared to conventional ACA. Electrical reliability of thermally conductive ACA flip chip joint under current stressing condition was also improved showing stable electrical conductivity of flip chip joints. The high current carrying capability and improved electrical reliability of thermally conductive ACA flip chip joint under current stressing test is mainly due to the effective heat dissipation by thermally conductive adhesive around Au stud bumps/ACA/PCB pads structure.