• KSBB Journal
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• v.6 no.3
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• pp.223-230
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• 1991

Using the shuttle vector pECCGl between Escherichia coli and Corynebacterium glutamicum and C. glutamicum strain JS231 grown in the medium supplemented with penicillin-G, which inhibits the formation of cross-links in the peptidoglycan of bacterial cell wall, various parameters involved in electroporation system including resistance, electric field strength, capacitance, DNA concentration, and cell density were investigated independently and optimized for the high efficiency transformation of coryneform bacteria. Using cells grown with 0.3U/ml of penicillin-G and harvested at A600 of 0.7-0.8, transformation efficiencies of 107-l08 transformants/$\mu\textrm{g}$ of DNA with Corynebcctertum glutamicum strain JS231 and wild type ATCC13032 were achieved under conditions of 12.5kV/cm of electric field strength, 400 ohms of resistance, $25\mu$F of capacitance, 3$\times$108 cells per transformation(1.2$\times$1010 cells/ml) and 100ng of plasmid DNA per transformation.

• ALGAE
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• v.22 no.3
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• pp.229-234
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• 2007

This study was to determine the extent of bacteria contamination and resistance to various antibiotics used commonly in microalgal culture. Seven different dose levels of chloramphenicol, dihydrostreptomycin sulphate, neomycin, penicillin G, streptomycin sulphate, penicillin G + streptomycin sulphate, and penicillin G + streptomycin sulphate + chloramphenicol were added to each culture of microalgae. The lethal effects on microalgae and bacteria were the highest in chloramphenicol and the lowest in penicillin G. The axenic culture of bacillariophyceae and dinophyceae was more difficult than that of chlorophyceae and haptophyceae because of their complicate external morphology. The efficient antibiotics and their concentrations for axenic cultures varied with microalgal species. The optimum quantity for antibiotic treatments were 2,000 ppm of dihydrostreptomycin for Chlorella ellipsoidea, neomycin 500 ppm of Isochrysis galbana and Heterosigma ahashiwo, hloramphenicol 500 ppm of Cyclotella didymus, and dihydrostreptomycin sulphate and neomycin 6,000 ppm of Thalassiosira allenii.

• YAKHAK HOEJI
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• v.40 no.1
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• pp.72-77
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• 1996

Penicillin sensor was manufactured by immobillizing penicillinase with glutaraldehyde on the $H^+$-selective membrane based on PVC-COOH-TDDA. This membrane was not inter fered by $K^+$ ion in Pc-G potassium salt. When enzyme was immobilized with glutaraldehyde, the PVC-COOH matrix was more effective than PVC matrix. Calibration curve calculated from Nernst equation was not linear. But potential was relative to concentration of Pc-G. And maximal potentiometric velocity was also relative to concentration of Pc-G. Therefore, it may be applied to Michaelis-Menten equation. The penicillin sensor was useful for determination of Pc-G at concentration of 0.1~10mM level.

• Journal of Life Science
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• v.27 no.10
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• pp.1145-1151
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• 2017

In this study, we report on a simple, efficient method for obtaining penicillin G amidase (PGA) from recombinant Escherichia coli using a formulation mixed with detergent and lysozyme. Research was conducted on the extraction efficiency of PGA from the periplasmic space in cells in terms of the type of detergent, detergent concentration, pH, reaction time, and temperature of permeabilization. The extraction yield of PGA in the formulated surfactant/lysozyme treatment was increased by approximately (55-65 U/ml) in comparison with that in the single surfactant treatment. The released PGA solution was concentrated and exchanged with buffer using an ultrafiltration (U/F) system. The yields of diatomite filtration, membrane filtration (M/F), and U/F were 69.7%, 93.8%, and 77.3%, respectively. A total of 212 KU of PGA was recovered. At the 25-L culture scale, the overall yield of extraction using the mixed surfactant/lysozyme method was 49.2%. The specific activity of extracted PGA was 11 U/mg in protein. The concentrated PGA solution was immobilized on microporous silica beads without further purification of PGA. The total immobilization yield of PGA on the resin was 48.7%, while the enzyme activity was 101 U/g. The immobilized PGA was successfully used to produce 6-APA from penicillin G. Our results indicated that a simple extraction method from periplasmic space in E. coli may be used for the commercial scale production of ${\beta}-lactam$ antibiotics using immobilized PGA.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1246-1256
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• 2015

Chlamydophila psittaci is an important intracellular pathogen. Persistent infection is an important state of the host-parasite interaction in this chlamydial infection, which plays a significant role in spreading the organism within animal populations and in causing chronic chlamydiosis and serious sequelae. In this study, a C. psittaci persistent infection cell model was induced by penicillin G, and real-time quantitative PCR was used to study the transcriptional levels of 10 C. psittaci genes (dnaA, dnaK, ftsW, ftsY, grpE, rpsD, incC, omcB, CPSIT_0846, and CPSIT_0042) in acute and penicillin-G-induced persistent infection cultures. Compared with the acute cultures, the penicillin-G-treated cultures showed a reduced chlamydial inclusion size and a significantly decreased number of elementary body particles. Additionally, some enlarged aberrant reticulate body particles were present in the penicillin-G-treated cultures but not the acute ones. The expression levels of genes encoding products for cell division (FtsW, FtsY) and outer membrane protein E encoding gene (CPSIT_0042) were downregulated (p < 0.05) from 6 h post-infection onward in the persistent infection cultures. Also from 6 h post-infection, the expression levels of DnaA, DnaK, IncC, RpsD, GrpE, and CPSIT_0846 were upregulated (p < 0.05); however, the expression level of OmcB in the persistent infection was< almost the same as that in the acute infection (p > 0.05). These results provide new insight regarding molecular activities that accompany persistence of C. psittaci, which may play important roles in the pathogenesis of C. psittaci infection.

• Korean Journal of Microbiology
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• v.32 no.3
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• pp.215-221
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• 1994

The complete nucleotide sequence of the cloned pga gene encoding the penicillin G acylase of Bacillus megaterium ATCC 14945 and its 5'- and 3'-flanking regions was determined. The sequence revealed only one large open reading frame (2,406 hp) of the penicillin G acylase (pga) gene. Upstream from ATG of the pga gene, there was a putative ribosome binding site, Shine-Dalgarno sequence. The promoter-like structure, - 10 and - 35 sequences, was also found. Following the stop codon, TAG, a structure reminiscent of the E. coli rho-independent transcription terminator was present. The amino acid sequence was deduced from the nucleotide sequence. The molecular mass of the polypeptide was 91,983 Da. There was a potential signal sequence in its amino-terminal region. A comparison of its deduced amino acid sequence with other characterized penicillin G acylases and the result of SDS-polyacrylamide gel electrophoresis of the purified enzyme showed that a precursor polypeptide of 92 kDa was processed into two dissimilar ${\alpha}$ and ${\beta}$-subunits of 25 and 61 kDa.

• Journal of Food Hygiene and Safety
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• v.8 no.1
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• pp.9-15
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• 1993

In an attempt to quantitate and qualitate residual antibiotics and antibacterial agents in meat simultaneously, we studied a gas chromatography-mass spectrometry(GC/M8) analysis. For a simultaneous analysis of penicillin G, chloramphenicol and thiamphenicol in meat, a simple and rapid clean-up procedure including extraction with 0.01 M EDTA-2Na Mcilvaine buffer (pH 4.0), defatting with n-hexane, and elution with 0.01M-methanolic oxalic acid from Bond Elute $C_{18}$ cartridge, and quantitation by selected ion monitoring (SIM) mode after derivatization was performed. The recoveries (%) of penicillin G, chloramphenicol and thiamphenicol (CV, %) at 1 ppm fortification level were 63.5 (7.6), 76.3 (8.1) and 84.7 (2.0), and the detection limits of those were 0.6, 0.085 and $0.084\;\mu\textrm{g}$ beef, respectively. This method using 81M mode allows excellent detection and quantitation of residual antibiotics and antibacterial agents in meat. Moreover, confirmation by a full scan electron impact mass spectrum is possible if residual level in the sample in above 1 ppm.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.11
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• pp.1604-1608
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• 2000

Penicillin antibiotics such as penicillin G, ampicillin and amoxicillin have been widely used in the pig industry to control salmonellosis, bacterial pneumonia, and urinary tract infections. Extensive use of antibiotics in veterinary clinics has resulted in tissue residues and bacterial resistance. To prevent unwanted drug residues entering the human food chain, extensive control measures have been established by both government authorities and industries. The demands for reliable, simple, sensitive, rapid and low-cost methods for residue analysis of foods are increasing. In this study, we established a rapid prediction test for the detection of pigs with unacceptable tissue residues of penicillins. The recommended therapeutic doses of three penicillins, penillin G (withdrawal time, 7 days), ampicillin (withdrawal time, 7 days) and amoxicillin (withdrawal time, 14 days), were administered to three groups of 20 pigs each. Blood was sampled before drug administration and during the withdrawal period. The concentration of penicillins in plasma, determined by a semi-quantitative ELISA, were compared to that of internal standard, 4 ppb, which corresponded to the Maximum Residue Limit in milk. The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine whether drug residues in pig tissues were negative or positive. That is, a B/Bs ratio less than 1 was considered residue positive, and larger than 1 negative. All 60 plasma samples from pigs were negative to three penicillins at pretreatment. Penicillin G could be detected in the plasma of the treated pigs until day 4 post-treatment and ampicillin until day 2, whereas amoxicillin could be detected until day 10 of its withdrawal period. The present study showed that the semi-quantitative ELISA could be easily adapted to detect residues of penicillin antibiotics (penicillin G, ampicillin and amoxicillin) in live pigs.

• Journal of Microbiology and Biotechnology
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• v.7 no.3
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• pp.194-196
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• 1997

Using the polyclonal antibody for Xanthomonas citri ampicillin acylase raised in Pseudomonas-free Balb/c mice, the immunochemical similarity of several types of penicillin acylases including Erwinia aroideae penicillin V acylase, Escherichia coli penicillin G acylase, Pseudomonas melanogenum and Acetobacter turbidans ampicillin acylases, and Pseudomonas cephalosporin acylase was examined. Among tested, only P. melanogenum ampicillin acylase showed the cross-reactivity with the antibody.

• Archives of Pharmacal Research
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• v.11 no.2
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• pp.163-165
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• 1988

Penicillin G potassium salt was transformed in high yield to 4-acetoxy-3-phenylacetaminoazetidinone by treatment of mercury acetate, ozonolysis and methanol in sequence.