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Enumeration of viable microbial cells by peroxyoxalate chemiluminescence method
오상곤,윤희주,차승희,정윤수,Oh, Sang-Kon,Youn, Hee-Ju,Cha, Seung-Hee,Chung, Yun-Su Korean Society for Biochemistry and Molecular Biol 1991 한국생화학회지 Vol.24 No.1
원핵 및 진핵세포의 전자전달계로부터 electron transport catalysts을 사용하여 과산화수소의 생성유도가 가능하였고 생성된 과산화수소는 peroxyoxalate 화학 발광반응을 이용하여 정량적으로 측정할 수 있었다. 따라서 과산화수소의 정량에 의한 이들 세포수의 측정이 가능하였다. 세포의 redox system으로부터 전자를 용존산소에 직접 전달하는 과산화수소의 생성능은 Escherichia coli의 경우 menadion이 phenazine ethosulfate와 phenazine methosulfate보다 우수한 것으로 나타났다. 그리고 과산화수소의 생성을 위한 menadione의 최적농도는 0.5 mM이었다. 0.5 mM menadione을 Escheichia coli에 혼합시 에 반응 10분내에 $3.5{\times}10^4\;cells/ml$까지 측정이 가능하였고 Saccharomyces cerevisiae는 반응시간 3분내에 $2.3{\times}10^4\;cells/ml$까지 측정 할 수 있었다. 그리고 Escherichia coli의 경우 반응 3분내에 측정가능한 세포수는 $3.5{\times}10^5\;cells/ml$이었다. Hydrogen peroxide was produced from viable cell by electron transport catalysts-mediated reaction. The concentration of hydrogen peroxide produced was determined by peroxyoxalate chemiluminescence method. Therefore, this method could be applied to the counting of bacteria and yeast cell. Among the tested electron transport catalysts, menadione was the most active catalyst for producing the hydrogen peroxide. And the optimum concentration for counting of viable cell was 0.5 mM. When 0.5 mM menadione was used, Escherichia coli cell ($8.1{\times}10^7\;cells/ml$) could produce $24.2\;{\mu}M$ hydrogen peroxide and the cell density above $3.5{\times}10^4\;cells/ml$ determined within 10 min. Saccharomyces cerevisiae cell density $2.3{\times}10^4\;cells/ml$ was determined within 3 min. The detectable minimum cell density of Esherichia coli was $3.5{\times}10^5\;cells/ml$ within 3 min.
Peroxyoxalate 화학발광법에 의한 세포수 측정에 관한 연구
오상곤,윤희주,차승희,정윤수 ( Sang Kon Oh,Hee Ju Youn,Seung Hee Cha,Yun Su Chung ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.1
Hydrogen peroxide was produced from viable cell by electron transport catalystsmediated reaction. The concentration of hydrogen peroxide produced was determined by peroxyoxalate chemiluminescence method. Therefore, this method could be applied to the counting of bacteria and yeast cell. Among the tested electron transport catalysts, menadione was the most active catalyst for producing the hydrogen peroxide. And the optimum concentration for counting of viable cell was 0.5 mM. When 0.5 mM menadione was used, Escherichia coli cell (8.1×10^7 cells/ml) could produce 24.2 μM hydrogen peroxide and the cell density above 3.5×10⁴cells/ml determined within 10 min. Saccharomyces cerevisiae cell density 2.3×10⁴cells/ml was determined within 3 min. The detectable minimum cell density of Esherichia coli was 3.5×10^5 cells/ml within 3 min.