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금강수역내에서 분리된 몇 종의 Ampicillin 내성균의 β-Lactamase 발현양상
이기성,고동규,최원창,문영길,박영식,최청일,최영길 ( Ki Sung Lee,Dong Kyu Ko,Won Chang Choi,Yong Keel Mun,Young Sik Park,Chung Il Choi,Yong Keel Choi ) 한국환경생물학회 1994 환경생물 : 환경생물학회지 Vol.12 No.1
In order to characterize the bacterial β-lactamase types occurred in Kum river area, this research was designed to analyze with identification upon ampicillin (Amp) resistant bacteria, and was investigated into relationships between β-lactamase expression (inducibility) and growth phase under ampicillin shift and with restriction patterns of R-plasmids. As the identification result upon 4 strains of Amp resistant bacteria, KL 601 is identified as Pasteurella maltocida, KL 602 as Enterobacter agglomerans, KL 609 as Pseudomonas maltophilia and KL 611 as Acinetobactor calcoaceticus, respectively. In order to investigate Amp effect upon growth, when the Amp resistant bacteria were cultivated in nutrient liquid LB medium, in LB medium added with Amp, or in LB medium in the manner of Amp shift, the changes in growth rate and lag phase could not be observed. However, upon the prototrophic strains, KL 602 and KL 609, examining Amp effect upon these two strains in minimal GM63 liquid media, even though there was no change of maximum growth yield, the latent lag phase become very lengthened. In nutrient medium, β-lactamase activities of these 4 strains were metabolically regulated in accordance with growth phase, on the other hand, in minimal medium were differently regulated in accordance with growth phase and with strains. It could be postulated that KL 609 strain has the M.W.24,000, β-lactamase type which E. coli comprise, however, that KL 601, KL 602 and KL 611 have the M.W.35,000, β-lactamase type which E. cloacae or C. freundii comprise. Through the whole growth phase, since Amp did not affect to phosholipid metabolism, there is no change in phospholipid compisition upon Amp effect. Main phospholipid are the phosphatidyl glycerol and cardiolipin in all 4 Amp resistant strains. So, it was very significant that there were no changes in phospholipid composition and in growth rate upon nutrient medium under Amp stress. In order to recognize whether Amp resistant strains comprise the plasmid or not, preparation of plasmids and restriction digestion with Bam H1 were carried out, KL 601 strain has about 30 Kb of plasmid, KL 609 and KL 611 strain have about 25 Kb of plasmid and KL 602 does not have the plasmid, however. Hence, it will be predicted strongly that β-lactamase gene in KL 602 might be encoded chromosomally. Moreover, considering the results upon the changes of β-lactamase acticity, upon β-lactamase inducibility and upon changes of periplasmic protein profile under Amp stress, its prediction is very recognizable.
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