Endang Purwati

Antimicrobial Properties and Lactase Activities from Selected Probiotic Lactobacillus brevis Associated With Green Cacao Fermentation in West Sumatra, Indonesia

Lactase activity is very important enzyme for Lactose Intolerant people. The lactic acid bacteria selected for use as Probiotic bacteria should be able to tolerate many organotoxic or heavy metal toxicities in intestine at least for 90 minute, attach to the epithelium, and grow in the lower intestinal tract before they can start providing any health benefits. The Green varieties of cacao fruits were used for cacao bean fermentation. The cacao bean covers with white pulp containing fructose, e.g. 42 mg/g, glucose 24 mg/g and sucrose 21 mg/g, as carbon source of Lactic Acid Bacteria (LAB). Screening of LAB was carried out after 24-36 h during spontaneous fermentation. This paper aims to find potential LAB resistant acid pH and producing potential antimicrobial and showing high protease/lactase activities. The medium of de Man, Ragosa, and Sharpe (MRS) were used to screen LAB, and 63 colonies were found. The screening of isolates is based on LAB survival growth in acid pH ranges (2.0; 2.5 and 3.0) and body temperature (37°C). For Antimicrobial experiments were used pathogen bacterial E. coli and Salmonela (Unand Collection) as indicator strain. Six isolate were conformed as strong antimicrobial and selected as potential for producing protease (lactase) using specific Triple Sugar Iron Agar (TSIA) medium. The isolate G3 and G6 were selected for further enzyme protease delivered with 2% Skim Milk as a protein subsrate. The results showed, the maximum amount of protease activity potential from isolate G6 in acid pH (3.0) were calculated as 0.0088 and protease activity 1.1795 Unit/mL, while the optimum of protease activity was found at pH (6.0) calculated protease activity 3.150 Unit/mL. This study could explain the possibility of using potential isolate G6 LAB as Probiotic for dairy or food Industry and supplement tablet. There is no report so far concerning this topic and potential isolate G6, with high antimicrobial or wide pH of protease. The study will continue to purify antimicrobial Bacteriocin and amino acid structure determination. The G6 isolate was 95% polymorphism with Lactobacillus brevis.

The Effect of Pediococcus pentosaceus on Stool Frequency, TNF-α Level, Gut Microflora Balance in Diarrhea-induced Mice

Background: Enteropathogenic Escherichia coli (EPEC) are pathogenic microorganisms causing inflammation and imbalanced gut microflora that may result in diarrhea. Pediococcus pentosaceus (P. pentosaceus) isolated from “dadih” (milk curd) are used as probiotics containing lactic acid bacteria (LAB), which are useful to improve the balance of intestinal microflora and inhibit the growth of pathogenic microorganisms. This study was aimed to recognize the effect of P. pentosaceus supplementation on stool frequency, tumor necrosis factor-α (TNF-α) and gut microflora balance in experimental mice with EPEC-induced diarrhea. Method : The study was conducted in 60 white mice (Mus muscullus) at Biomedical Laboratory, Biotechnology/Production and Animal Husbandry Technology Institute, University of Andalas, Padang in April 2012. The frequency of stool, TNF-α level and microflora balance of the mice were measured before and after the EPEC-induced diarrhea and following the administration of antibiotics. Statistical analysis was performed using ANOVA and Duncan test. Results: The highest mean stool frequency was found in positive control group, i.e. 55 times, which was reduced significantly after 12-hour P. pentosaceus supplementation in a dose of 2 x 108 cfu/g into 18 times. The mean TNF-α level in positive control group was 128.17 pg/mL that lowered significantly to 48.0 pg/mL. The highest mean total number of LAB was 97.0 x 107 cfu/g, which was significantly different from positive control group of 7 x 107 cfu/g. Conclusion: P. pentosaceus supplementation in a dose of 2 x 108 cfu/g may reduce the stool frequency, lower TNF-α and improve the gut microflora balance following 12-hour supplementation in diarrhea-induced mice. Keywords: Pediococcus pentosaceus, TNF-α, diarrhea, EPEC, gut microflora