[Objective] The aim was to study if the fermentation broth of Kombucha has protein with antimicrobial activity. [Method] The effects of different cultivation time on cell concentration, pH, total protein concentration...[Objective] The aim was to study if the fermentation broth of Kombucha has protein with antimicrobial activity. [Method] The effects of different cultivation time on cell concentration, pH, total protein concentration and inhibition zone were studied. The fermentation broth of Kombucha on the sixth day was adjusted to different pH. Fermentation broth treated with protease was put into the plates of Escherichia co/i, Bacillus cereue and Staphylococcus aureus as control, as well as ampenicilin. The zone of broth inhibition was measured. [ Result] The concentration of general protein was of positive relevance to the antimicrobial activity of Kombucha fermentation broth. The zone of inhibition decreased with rising pH and was the lowest when pH was 7. Later, as pH increasing, it enlarged. The inhibition effect of processed protease reduced remarkably. [ Conclusion] The fermentation broth of Kombucha revealed antimicrobial proteins.展开更多
Objective: To construct mammalian cell expression vectors for Mac-1 with CFP and YFP and apply FRET to study the dimerization and function of CD11 b( Mac-1 α subunit) and CD18(Mac-1 β subunit). Methods: The mammalia...Objective: To construct mammalian cell expression vectors for Mac-1 with CFP and YFP and apply FRET to study the dimerization and function of CD11 b( Mac-1 α subunit) and CD18(Mac-1 β subunit). Methods: The mammalian cell expression vector for CD11b fused with CFP at the carboxyl terminal was constructed to create recombinant plasmid of pCD11b-CFP. Then pCD11b-CFP was co-transfected with pYFP-CD18 into CHO cell, a fibroblast like cell line, as a target cell within which there are some signal pathways involved in inflammatory stimulation but without endogenous Mac-1. Then CHO cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were selected by Western blot and laser scanning confocal microscope. Results: The cyan and yellow fluorescence in co-transfected positive CHO cells were observed under a fluorescence microscope. CHO-Mac-1-FP cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were obtained as demonstrated by Western blot successfully. The adhesive activity of CHO-Mac-1-FP cells with CHO-1CAM-1 cells was increased markedly by treatment with PMA, suggesting the translocation of GD11b-CFP and YFP-CD18 to the plasma membrane in CHO-Mac-1-FP cells and dimerization of CD11b-CFP and YFP-CD18 just as the function of the wild type Mac-1. Conclusion: CHO-Mac-1-FP cells with adhesive activity are established successfully, thus CHO-Mac-1-FP cells may be useful for the study of Mac-1 by FRET and for other purposes.展开更多
基金Supported by Scientific Research Fund of Beijing Educational DepartmentBeijing Higher School Personnel Teaching Program
文摘[Objective] The aim was to study if the fermentation broth of Kombucha has protein with antimicrobial activity. [Method] The effects of different cultivation time on cell concentration, pH, total protein concentration and inhibition zone were studied. The fermentation broth of Kombucha on the sixth day was adjusted to different pH. Fermentation broth treated with protease was put into the plates of Escherichia co/i, Bacillus cereue and Staphylococcus aureus as control, as well as ampenicilin. The zone of broth inhibition was measured. [ Result] The concentration of general protein was of positive relevance to the antimicrobial activity of Kombucha fermentation broth. The zone of inhibition decreased with rising pH and was the lowest when pH was 7. Later, as pH increasing, it enlarged. The inhibition effect of processed protease reduced remarkably. [ Conclusion] The fermentation broth of Kombucha revealed antimicrobial proteins.
基金Supported by National Natural Science Foundation of China (No.30000068, 39730210)
文摘Objective: To construct mammalian cell expression vectors for Mac-1 with CFP and YFP and apply FRET to study the dimerization and function of CD11 b( Mac-1 α subunit) and CD18(Mac-1 β subunit). Methods: The mammalian cell expression vector for CD11b fused with CFP at the carboxyl terminal was constructed to create recombinant plasmid of pCD11b-CFP. Then pCD11b-CFP was co-transfected with pYFP-CD18 into CHO cell, a fibroblast like cell line, as a target cell within which there are some signal pathways involved in inflammatory stimulation but without endogenous Mac-1. Then CHO cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were selected by Western blot and laser scanning confocal microscope. Results: The cyan and yellow fluorescence in co-transfected positive CHO cells were observed under a fluorescence microscope. CHO-Mac-1-FP cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were obtained as demonstrated by Western blot successfully. The adhesive activity of CHO-Mac-1-FP cells with CHO-1CAM-1 cells was increased markedly by treatment with PMA, suggesting the translocation of GD11b-CFP and YFP-CD18 to the plasma membrane in CHO-Mac-1-FP cells and dimerization of CD11b-CFP and YFP-CD18 just as the function of the wild type Mac-1. Conclusion: CHO-Mac-1-FP cells with adhesive activity are established successfully, thus CHO-Mac-1-FP cells may be useful for the study of Mac-1 by FRET and for other purposes.
