IP Library Granted Patent US 9,596,861
Granted Patent B2
US 9,596,861 · App. 14/576,411 · Granted Mar 21, 2017

Method of producing partially purified extracellular metabolite products from

Inventors: Muhammed Majeed (Edison, NJ); Kalyanam Nagabhushanam (East Windsor, NJ); Sivakumar Arumugam (Bangalore, IN); Furqan Ali (Bangalore, IN)
Assignee: SAMI LABS LIMITED
A01N63/02A01N31/08A01N37/12C12N1/20C12R1/07C12R1/085C12R1/19C12R1/385C12R1/42C12R1/445C12R1/45C12R1/46
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Quick Facts
Patent No.
US 9,596,861
App. No.
14/576,411
Granted
Mar 21, 2017
Kind
B2
Abstract

Method of extracting a partially purified extracellular metabolite composition from Bacillus coagulans MTCC 5856 and its antimicrobial effects thereof are described.

Claims (11)

1. A purification method for producing partially purified extracellular metabolite preparation from the probiotic bacterial strain Bacillus coagulans strain MTCC 5856), said purification method comprising the steps of:

a) Inoculating a culture of Bacillus coagulans MTCC 5856 into 1.0 liter of Glucose Yeast Extract Acetate broth medium or MRS broth containing 0.5% Tween 80 or Corn steep powder media to initiate bacterial fermentation;

b) Allowing the bacterial fermentation in the inoculated medium of step a to proceed for 24-48 h at 37° C. with 120 rpm;

c) Centrifuging the fermentation broth of step b at 4000-7000 rpm and collecting supernatant;

d) Concentrating supernatants 10 fold by using rotary evaporator at 50° C. of step c;

e) Adding 150 ml of chilled acetone drop by drop to 100 ml of tenfold concentrated supernatants of step d, followed by mixing to form supernatants-acetone mixture;

f) Incubating the supernatants-acetone mixture of step e at 0° C. for 30 minutes followed by centrifuging at 7000-8000 rpm to collect a pellet;

g) Discarding the pellet obtained in step f and collecting 60% (v/v) acetone saturated supernatant (˜200 ml);

h) Concentrating the acetone saturated supernatant in step g to 50 ml by rotary evaporator;

i) Adjusting the pH of the supernatant of step h 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 the supernatant of step I to obtain partially purified extracellular metabolite preparation from the probiotic bacterial strain Bacillus coagulans MTCC 5856.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 12, 2016
From: MAJEED, MUHAMMED, DR; NAGABHUSHANAM, KALYANAM, DR; ARUMUGAM, SIVAKUMAR, DR; ALI, FURQAN, DR
To: SAMI LABS LIMITED
Reel/Frame 037484/0268 →
Continuity (2)
Provisional Application 61920567 · Dec 24, 2013
Related Publication 20150173374A1 · Jun 25, 2015