METHOD OF PRODUCING PARTIALLY PURIFIED EXTRACELLULAR METABOLITE PRODUCTS FROM BACILLUS COAGULANS AND BIOLOGICAL APPLICATIONS THEREOF

摘要

Disclosed is a method for producing partially purified extracellular metabolite preparation from the probiotic bacterial strain Bacillus coagulans SBC-37 (Deposited in the Microbial Type Culture Collection and Gene Bank as strain number MTCC 5856) exhibiting 99% genetic homology with the known bacterial strains Bacillus coagulans ATCC 31284, Bacillus coagulans NBRC 3887 and Bacillus coagulans ATCC 7050. Also disclosed is the anti-microbial profile of said extracellular metabolite preparation against a panel of microbial pathogens, including synergistic anti-microbial effects of preparation when combined with a synergistic preservative blend comprising from about 61% w/w of thymol, about 38% of monolaurin and about 1% w/w of magnolol obtained from supercritical fluid extracts of Magnolia officinalis. The extracellular metabolite preparation alone or the combination of said extracellular metabolite preparation and preservative blend also inhibits microbial biofilm formation in a synergistic manner.

主权项

1. A purification method for producing partially purified extracellular metabolite preparation from the probiotic bacterial strain Bacillus coagulans SBC37-01 (Deposited in the Microbial Type Culture Collection and Gene Bank and was assigned the strain number MTCC 5856) exhibiting 99% genetic homology with the known bacterial strains Bacillus coagulans ATCC 31284, Bacillus coagulans NBRC 3887 and Bacillus coagulans ATCC 7050, said purification method comprising the steps of:
a) Inoculating a culture of Bacillus coagulans MTCC 5856 exhibiting 99% genetic homology with the known bacterial strains Bacillus coagulans ATCC 31284, Bacillus coagulans NBRC 3887 and Bacillus coagulans ATCC 7050 into 1.0 liter of Glucose Yeast Extract Acetate broth medium or MRS broth containing 0.5% tween 80 or Corn steep powder media; b) Allowing the fermentation in the inoculated medium of step 1 to proceed for 24-48 h at 37° C. with 120 rpm; c) Centrifuging the fermentation broth of step 2 at 4000-7000 rpm; d) Concentrating supernatants 10 fold by using rotary evaporator at 50° C. of step 3. e) Adding 150 ml of chilled acetone drop by drop to 100 ml of tenfold concentrated supernatants of step 4, followed by mixing; f) Incubating the mixture of step 5 at 0° C. for 30 minutes followed by centrifuging at 7000-8000 rpm; g) Discarding the pellet obtained in step 6 and collecting 60% acetone saturated supernatant (˜200 ml). h) Concentrating the acetone saturated supernatant in step 7 to 50 ml by rotary evaporator. i) Adjusting the pH to 5.0 by using 4N HCl, filtered (0.22 micron; Millex, Millipore, India) and stored at −20° C. till further use. j) Freeze drying/spray drying/tray drying the supernatant of step 8.