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Bacillus subtilis M-15 strain for degrading gossypol, fungicide and application

A Bacillus subtilis, M-15 technology, applied in the direction of bacteria, microbe-based methods, microbes, etc., can solve the problems of non-genus and unusable fungi

Pending Publication Date: 2022-01-25
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the types of detoxification agents used in cottonseed meal fermentation are mainly concentrated in molds and yeasts, or composite strains mainly composed of molds and yeasts. A few studies have mentioned the use of bacteria such as lactic acid bacteria, but some of them do not belong to our country The strains announced by the Ministry of Agriculture can be used to produce microecological preparations, and fungi are generally not available for large-scale fermentation of cottonseed meal for industrial production

Method used

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  • Bacillus subtilis M-15 strain for degrading gossypol, fungicide and application
  • Bacillus subtilis M-15 strain for degrading gossypol, fungicide and application
  • Bacillus subtilis M-15 strain for degrading gossypol, fungicide and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The present embodiment provides a kind of screening process of M-15 bacterial strain:

[0033] The media used are as follows:

[0034] Beef extract peptone medium (NA): 5.0g beef extract, 10.0g peptone, 5.0g NaCl, 18.0g agar, 1000mL distilled water, pH 7.2-7.4.

[0035] Liquid beef extract peptone medium (NB): 5.0g beef extract, 10.0g peptone, 5.0g NaCl, 1000mL water, pH 7.2-7.4.

[0036] Gossypol carbon source medium: gossypol 5.0g, (NH 4 ) 2 SO 4 2.0g, MgSO 4 ·7H 2 O 0.2g, NaH 2 PO 4 ·H 2 O0.5g, CaCl 2 2H 2 O 0.1g, K 2 HPO 4 0.5g, 15.0g agar, 1000mL distilled water, pH 7.2-7.4.

[0037] Gossypol carbon source culture medium: gossypol 5.0g, beef extract 1.0g, peptone 2.0g, (NH 4 ) 2 SO 4 2.0g, MgSO 4 ·7H 2 O0.2g, NaH 2 PO 4 ·H 2 O 0.5g, CaCl 2 2H 2 O 0.1g, K 2 HPO 4 0.5g, distilled water 1000mL, pH 7.2-7.4.

[0038]Acid resistance test medium: beef extract 0.5%, peptone 1.0%, sodium chloride 0.5%, pH adjusted to 4.5;

[0039] Inorganic nitrog...

Embodiment 2

[0090] The present embodiment provides a bacterial preparation prepared by the Bacillus subtilis M-15 bacterial strain, and the process is as follows:

[0091] 1) Inoculate the screened M-15 strain into NA medium for culture to obtain slant seeds;

[0092] 2) Take the slant seeds and inoculate them into the NB medium for 14 hours of cultivation to obtain the culture solution;

[0093] 3) inoculate the culture solution into the fermentation medium with 10% inoculum amount, and ferment to obtain the fermentation solution; wherein, the fermentation temperature is 30°C, the fermentation time is 60h, the liquid loading volume is 120ml / 250ml, and the ventilation volume is 1 : 0.8V / V; every liter of said fermentation medium comprises: sucrose 5.0g, nitrogen source 10g, MgSO 4 ·7H 2 O 0.2g, NaH 2 PO 4 ·H 2 O 0.5g, CaCl 2 2H 2 O0.1g, K 2 HPO 4 0.5 g, FeCl 3 0.2 g, distilled water 1000mL, pH 7.2-7.4;

[0094] 4) Dry the fermentation broth under the conditions of an air inlet ...

Embodiment 3

[0096] This embodiment provides the screening process of culture medium, fermentation conditions and inoculum size in the production process of bacterial agents;

[0097] Media used:

[0098] Basic liquid fermentation medium: carbon source 5.0g, nitrogen source 10g, MgSO 4 ·7H 2 O 0.2g, NaH 2 PO 4 ·H 2 O 0.5g, CaCl 2 2H 2 O 0.1g, K 2 HPO 4 0.5 g, FeCl 3 0.2 g, distilled water 1000mL, pH 7.2-7.4.

[0099] Fermentation equipment

[0100] 1m 3 Seed tank, 20m 3 Fermentation tanks, other fermentation equipment. Related 20m 3 The fermenter process test was carried out at Hebei Zhongbang Biotechnology Co., Ltd.

[0101] Preparation of detoxified Bacillus bacteria agent from cottonseed meal

[0102] Determination of the Optimal Age of the Strains

[0103] Fresh slant seeds of the test strain were inoculated with NB medium for culture, samples were taken regularly, the fermentation broth was diluted 10 times, and the relative bacterial concentration (light absorption v...

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Abstract

The invention provides a bacillus subtilis M-15 strain for degrading gossypol, the strain is Bacillus subtilis M-15, the strain is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation date is September 26, 2021, the preservation number is CGMCC No.23483, and the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing; and the bacillus subtilis is classified and named as Bacillus subtilis.

Description

technical field [0001] The invention relates to the technical field of bacterial strain screening and microbial agent preparation, and more specifically relates to the Bacillus subtilis M-15 strain with the function of degrading gossypol, bacterial agent and application. Background technique [0002] At present, the feed protein of my country's animal husbandry industry mainly comes from soybean meal. As the price of soybean meal rises, the development of other cake protein feed resources is one of the ways to solve the shortage of protein resources in my country. [0003] Cotton meal is rich in protein, and the crude protein content of cottonseed cake and meal made from completely shelled cottonseed can reach 40%, which is comparable to that of soybean cake; cottonseed produced by directly pressing oil from unhulled cottonseed The crude fiber content of the cake is 16%-20%, the crude protein content is 20%-30%, and 17 kinds of amino acids can be obtained after the cotton me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/125
CPCC12N1/20A23L5/28
Inventor 李佳朱宝成高同国王伟
Owner HEBEI AGRICULTURAL UNIV.
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