Semi-purification and characterization of bacteriocin from Ruminococcus albus 7
Date Issued
2008
Date
2008
Author(s)
Chen, I-Hung
Abstract
Our research was set up to find more detailed characteristics of the bacteriocin from R. albus 7 and its potential as alternative of antibiotics. We also tried to establish reliable production and semi-purification procedures. In our results, R. albus 7 could be cultured in the medium without ruminal fluid to produce bacteriocin effectively against five pathogens (Enterobacter, Bacillus, Pseudomonas, Samonella, Staphylococcus). MIC50 and MIC90 of R. albus 7 bacteriocin were as low as 0.75 mg/mL and 6.5 mg/mL, respectively. The bacteriocin is highly released after 24 hr of incubation. Inclusion of 3μM 3-phenylpropanoic acid（PPA）and 0.2 % Tween 80 in culturing medium improved 2.5 times production of bacteriocin. Native-gel electrophoresis showed that bacteriocin of R. albus 7 had a molecular weight about 33 kDa. Semi-purification procedure resulted in 23% recovery rate of bacteriocin by using the DEAE column in FPLC system. The bacteriocin of R. albus 7 could be destroyed by pepsin, protease, and pancreatin. It also could be inactivated after heating at 65℃for 1 hr. Simulated in vitro avian digestion decreased the antagonistic activity by 74.7%. But, addition 1% BSA could retain 13% more antagonistic activity.n conclusion, the bacteriocin of R. albus 7 is a heat-labile small protein. PPA and Tween 80 are recommended to be included in its production culture medium. DEAE column was suitable for its semi-purification. The bacteriocin of R. albus 7 has the chance as an antibiotic alternative with protectant added.
Subjects
bacteriocin
ruminal bacteria
antibiotic alternative
Type
thesis
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