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Composition comprising fermentation products from bacillus subtilis

A technology of Bacillus subtilis and fermentation products, applied in the direction of fermentation, biocide-containing paint, microbial-based methods, etc., can solve the problems of microbial contamination of products, prolongation of time period, etc., and achieve the goal of preventing pollutants and/or microbial contamination Effect

Inactive Publication Date: 2014-12-24
DUPONT NUTRITION BIOSCIENCES APS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Also, in use, the packaging may be opened and / or removed for an extended period of time before the product is fully consumed or applied
For example, in some dry products, such as dry foods (e.g., pet food), the time period between first opening of the product by the user and final consumption may be prolonged, allowing microbial contamination of the product

Method used

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  • Composition comprising fermentation products from bacillus subtilis
  • Composition comprising fermentation products from bacillus subtilis
  • Composition comprising fermentation products from bacillus subtilis

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0624] Example 1 - Preparation of antibacterial samples

[0625] Growth of Antimicrobial Strains

[0626] Strains: Bacillus subtilis 22C-P1 (DCS 1579), 15A-P4 (DCS 1580), 3A-P4 (DCS 1581), LSSAO1 (DCS 1582), ABP278 (DCS 1583) and BS18 (DCS 1584) from blood agar Resuscitation from deep-frozen stock cultures. Isolated colonies from each culture were streaked on CASO agar and incubated aerobically at 32°C for 24 hours. One colony of each culture was transferred to 10 mL of CASO broth in a 50 mL SARSTEDT tube and incubated for 24 hours at 32°C with tilting shaking at 130 rpm. 0.5 mL of the growth culture was transferred to 50 mL of CASO broth in a 250 mL Erlenmeyer flask and incubated at 32°C with shaking at 130 rpm for 24 hours.

[0627] Preparation of Antibacterial Supernatant Samples

[0628] Fully grown cultures were centrifuged twice at 10.000 xg for 10 minutes each. The supernatant was filter sterilized (using vacuum) and the filtrate was used immediately.

example 2

[0629] Example 2 - Inhibition Range Determination

[0630] The well diffusion assay was used to assess the extent of inhibition of cell-free supernatants (CFS) prepared in Example 1 against various target microorganisms (Table 1). Prepare one plate for each indicator microorganism. 30 mL of molten agar medium containing 3 mL of 2M sodium phosphate (pH 6.5) was inoculated with 150 μl of the fully grown overnight culture and mixed well. Pour the suspension into the omnitray and let it settle for 30 minutes. Six wells were cut in the agar and left to dry open for another 30 minutes in the LAF stand. Each duplicate well was filled with 100 μl of supernatant as previously prepared and incubated at the corresponding temperature, time and conditions as shown in Table 1. After the incubation time, the diameter of the inhibition zone was assessed and divided into inhibitory populations. Activity is marked as "+" for zone diameters up to 10 mm including the aperture, "++" for zone...

example 3

[0645] Example 3 - Sensitivity of activity to heat treatment and various pH

[0646] 30 mL of CFS for each strain was divided into 6 aliquots of 5 mL and the pH was adjusted to pH 4, 5, 6, 7, 8 or 9 using 5M NaOH or 5M HCl. Each pH adjusted 5 mL aliquot was filter sterilized, divided into 5 aliquots of 0.8 mL and stored at 4°C until use.

[0647]Heat treatment was applied as described in Table 5 for each CFS. Six aliquots (one for each pH value) were heat treated at 72°C for 15 seconds. Temperature was monitored using a temperature probe through a hole in the lid in an eppendorf tube filled with 0.8 mL of CASO broth. 15 seconds were counted from the moment the temperature reached 72°C. Another set of 6 aliquots was heat treated at 100°C for 10 minutes. Temperature was monitored using a temperature probe through a hole in the lid in an eppendorf tube filled with 0.8 mL of CASO broth. Ten minutes were counted from the moment the temperature reached 95°C. Six aliquots wer...

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Abstract

The present invention relates to anti-contaminant composition comprising a cell-free fermentation product of one or more Bacillus subtilis strains (e.g., selected from the group consisting of: 22C-P1, 15A-P4, 3A-P4, LSSA01, ABP278, BS 2084 and BS18); wherein said fermentation product comprises one or more compounds selected from the group consisting of: a lipopeptide, a polyketide, a bacillibactin, a bacilysin, an anticapsin, a plantazolicin, a LCI, a homologue of a plantazolicin and a homologue of a LCI. In addition, the present invention further relates to methods of preparing the compositions, methods of using the composition, products comprising the composition and uses thereof.

Description

[0001] priority statement [0002] This application claims priority to US Patent Application No. 61 / 601,154, filed February 21, 2012, which is incorporated by reference in its entirety. technical field [0003] The present invention relates to anti-pollution compositions, processes for their preparation and their use to prevent microbial contamination of products such as food, surface coating materials and agricultural products. In particular, the present invention relates to anti-pollution compositions comprising B. subtilis strains such as 22C-P1, 15A-P4, 3A-P4, LSSA01, ABP278, BS 2084 and BS18 fermentation product. Background technique [0004] Microbial contamination of products is a problem in several industries. [0005] For example, in the paint industry, water-based paints are susceptible to microbial contamination (eg, spoilage) when wet. Such contamination can lead to discoloration, outgassing, malodor, loss of viscosity, stringing (ie, slime) and phase separat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P1/04A23L1/30C12R1/125A01N63/22A23K20/195
CPCC12N1/20A23L3/3463A23L3/3472A23L3/3526A23K10/12A23K20/111A23K20/147A23K20/158A23K20/10A23K50/40A01N63/22C12R2001/125C12N1/205A01N2300/00A23L3/34635A23V2002/00C09D5/14
Inventor G. H. 维伯T. 麦金德C. 本菲尔德A. A. 海博德R. J. 兰德姆P. 查诺斯G. R. 斯拉古萨M. J. 亨特
Owner DUPONT NUTRITION BIOSCIENCES APS
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