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A kind of fermentation broth and fermentation method for extracellular antibacterial protein produced by Bacillus amyloliquefaciens sz-60

A technology for desolvating amyloliquefaciens and fermentation methods, which is applied in the field of Bacillus amyloliquefaciens SZ-60 fermentation broth for extracellular protein production, can solve problems such as soil pesticide residues, soil microecological imbalance, etc., and achieves improvement of bacteriostatic efficiency and protein content. Improved, good control effect

Active Publication Date: 2021-09-28
JILIN AGRICULTURAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the control of ginseng botrytis is still dominated by chemical control, and the spread of the disease is controlled through the application of mancozeb and procymidone, but long-term large-scale use of chemical pesticides will cause disease resistance, soil pesticide residues, and soil micro-ecological imbalance wait for some questions

Method used

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  • A kind of fermentation broth and fermentation method for extracellular antibacterial protein produced by Bacillus amyloliquefaciens sz-60
  • A kind of fermentation broth and fermentation method for extracellular antibacterial protein produced by Bacillus amyloliquefaciens sz-60
  • A kind of fermentation broth and fermentation method for extracellular antibacterial protein produced by Bacillus amyloliquefaciens sz-60

Examples

Experimental program
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Effect test

Embodiment 1

[0068] Bacillus amyloliquefaciens SZ-60 is preserved by the China General Microorganism Culture Collection and Management Center, and Botrytis cinerea Pers.ex Fr. is preserved by the State Key Laboratory of Ecological Restoration and Ecosystem Management jointly established by Jilin Agricultural University.

[0069] The activation of Bacillus amyloliquefaciens SZ-60 uses beef extract peptone medium (beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 16.0g, deionized water 1000mL, pH 6.8-7.0); Beef extract peptone culture solution (beef extract peptone culture medium without agar); the preparation of fermented seed liquid adopts NBYM fermentation liquid (beef extract 4.0g, peptone 9.0g, yeast extract 6.5g, malt extract 8.0g, glucose 3.0 g, MgSO 4 10.0mg, deionized water 1000mL, 6.8-7.2); the activation of Botrytis cinerea uses PDA medium (potato starch 5.0g, glucose 20.0g, agar 22.0g, deionized water 1000mL, pH 7.0-7.2). The 8 kinds of fermented liquids involved in this embodi...

Embodiment 2

[0110] Preparation of Bacillus amyloliquefaciens SZ-60 Bacterial Suspension

[0111] Activate the Bacillus amyloliquefaciens SZ-60 test tube species in beef extract peptone medium, culture in an artificial climate box at 37°C for 24 hours, pass passage 3 times, pick a single colony and inoculate it in a 250mL triangular flask containing 50mL beef extract peptone culture medium, 32 ℃, 180 rpm constant temperature shaking culture for 18-24h, the obtained bacterial culture solution was centrifuged twice at 4℃, 4500 rpm, 20min, the supernatant was discarded, and the obtained bacteria were made into bacteria-containing bacteria by using the hemocytometer method. The amount is about 10 8 CFU / mL bacterial suspension, set aside.

[0112] Calculation of the bacterial content of the bacterial suspension: using the hemocytometer method. Spot 10 μL of bacterial fermentation broth on the hemocytometer, record the bacterial cells in the middle, upper left, lower left, upper right, and low...

Embodiment 3

[0115] Preparation of NB60YF microbial preparation

[0116] NB60YF microbial preparation contains the whole fermentation broth culture of Bacillus amyloliquefaciens SZ-60, the preparation method is as follows:

[0117] (1) Draw 5mL of the SZ-60 bacterial suspension of Example 2, inoculate it in a 500mL Erlenmeyer flask containing 125mL NBYM fermentation broth, cultivate it at a constant temperature of 33°C and 165 rpm for 12h with constant temperature shaking, and then test the SZ-60 bacterial strain every 2h The fermentation quality of the fermentation liquid is tested until the growth of the fermentation liquid is relatively full and spores are about to appear, then the fermented seed liquid is obtained, and the fermentation time is 24-28 hours;

[0118] (2) Inoculate the fermented seed liquid with an inoculation amount of 4% in a 500mL Erlenmeyer flask containing 200mL NB60Y fermented liquid, culture on a shaking table, ferment at a temperature of 30°C, rotate at a speed of...

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Abstract

The invention discloses a fermentation liquid for extracellular protein produced by Bacillus amyloliquefaciens SZ‑60 and a fermentation method thereof. For every 1000mL of deionized water in the fermentation broth, it contains 10-20g of soluble starch, 10-20g of soybean powder, 2-7g of NaCl, MnSO 4 2‑7mg, Ca(HCO 3 ) 2 5‑15mg, FeSO 4 1‑4mg; the pH of the fermentation broth is 6.5‑7.5. The invention optimizes and understands the combination of fermentation and culture conditions for producing protein antibacterial substances of Bacillus amyloliquefaciens SZ‑60, and provides technical support and theoretical guidance for the industrial production of Bacillus amyloliquefaciens SZ‑60 strains and the acquisition of high-yielding protein antibacterial substances. The NB60YF microbial preparation fermented by Bacillus amyloliquefaciens SZ‑60 has a good control effect on gray mold caused by Botrytis cinerea.

Description

technical field [0001] The invention belongs to the technical field of fermentation process of extracellular antibacterial protein of Bacillus amyloliquefaciens SZ-60, and in particular relates to a fermentation liquid for producing extracellular protein of Bacillus amyloliquefaciens SZ-60 and a fermentation method thereof. Background technique [0002] Ginseng (Panax ginseng Meyer) is a perennial root medicinal plant of Araliaceae Panax genus, which is widely planted in China, South Korea, Russia, North Korea and other countries. Northeast China is the main production area of ​​Chinese ginseng, and it is planted in a large area at the foot of Changbai Mountain. Due to the special geographical location, ginseng botrytis caused by Botrytis cinerea Pers.ex Fr. occurs frequently. At present, the control of ginseng botrytis is still dominated by chemical control, and the spread of the disease is controlled through the application of mancozeb and procymidone, but long-term large...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/07
CPCA01N63/00C12N1/20C12P21/00
Inventor 孙卓杨利民韩梅韩忠明程林吕泽良
Owner JILIN AGRICULTURAL UNIV
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