|Development of method to quantify extracellular carbohydrate complexes produced by Escherichia coli O157H7|
|Year of publication||2004년 이전|
|Author||Ryu, J.H., & L. R. Beuchat.|
|Publication in journal||Journal of Applied Microbiology 2003, 95, 1304–1314|
|Status of publication||Published|
Aims: The aim of this study was to optimize conditions to separate extracellular carbohydrate complexes (ECC) produced by Escherichia coli O157:H7 and to standardize the amount of ECC produced on a per cell basis.
Methods and Results: ECC fraction I was removed from E. coli O157:H7 cells produced on tryptic soya agar and lettuce juice agar by centrifugation. To remove ECC fraction II, cells were heated at 100C for 10 min, then centrifuged. The sum of ECC fractions I and II was considered as the total ECC produced by E. coli O157:H7. A correlation between cell mass and turbidity (O.D.750nm) of cell suspensions was determined. Cell mass has a linear relationship (R2 = 0.93) with turbidity of cell suspensions from which ECC is removed. The amount of ECC produced on a per cell basis was calculated by dividing total amount of ECC (l g ml-1) produced by the turbidity (O.D.750nm) of heated cell suspension after removal ECC fractions I and II.
Conclusions: A method for separating ECC from cells of E. coli O157:H7 has been developed and conditions have been optimized. A standard method to estimate the amount of ECC produced on a per cell basis was also developed.
Signiﬁcance and Impact of the Study: Using these procedures to prepare extract of ECC from E. coli O157:H7 and to standardize values, production of ECC on a per cell basis can be estimated and a comparison of the amount of ECC produced by the pathogen grown under different environmental conditions can be accurately measured.