Composite antibacterial peptide produced by bacillus natto NT-6 and production method thereof

A technology of Bacillus natto and NT-6, which is applied in the biological field, can solve the problems of no identification and no literature on the identification of antibacterial substances of Bacillus natto, and achieve high safety effects

Inactive Publication Date: 2012-01-18
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some researchers believe that the antibacterial substance produced is 2,6-pyridinecarboxylic acid, and some people think that it is polymyxin or bacitracin or antibacterial protein, but there is no identification data (Ji Ning et al., Food Research and Development, 2006, 27 (1): 138-142); Cao Xiaohong et al. Bacillus The antibacterial substances produced by the natto TK-1 strain were studied, and a cyclic lipopeptide surfactin with 15 carbons on the fatty acid chain with a molecular weight of 1036Da was found (Cao Xiaohong et al., China Biotechnology Journal, 2009, 29 (2):54-58)
In addition, there are no other relevant literatures on the identification of antibacterial substances of Bacillus natto, especially no literature reports on the simultaneous production of more than 20 antibacterial lipopeptide homologues by one strain

Method used

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  • Composite antibacterial peptide produced by bacillus natto NT-6 and production method thereof
  • Composite antibacterial peptide produced by bacillus natto NT-6 and production method thereof

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Effect test

Embodiment 1

[0027] After expanded cultivation of seeds, submerged fermentation was carried out in a fermenter at 28-35°C for 30-48 hours to accumulate antibacterial metabolites. The fermentation liquid was centrifuged to remove bacteria, and then HCl was added to the supernatant to adjust the pH to 2. Centrifuge after placing, the precipitate is further extracted with 3-5 times of methanol or ethanol, the extract is adsorbed by excipients and dried to obtain the finished product of antibacterial peptide, which is determined by high performance liquid chromatography, and the content of antibacterial peptide is not less than 100g / kg.

Embodiment 2

[0029] Bacillus natto ( Bacillus natto ) After the NT-6 bacterial strain is expanded through the above-mentioned seed medium, carry out liquid submerged fermentation in the above-mentioned liquid fermentation medium in a fermenter at 28-35° C., accumulate antibacterial metabolites for 30-48 hours, and remove the bacteria through centrifugation of the fermentation liquid. Then the supernatant is concentrated and dried to obtain the finished product of the antimicrobial peptide, and the content of the antimicrobial peptide is not less than 100 g / kg as determined by high performance liquid chromatography.

Embodiment 3

[0031] Bacillus natto ( Bacillus natto ) After the NT-6 bacterial strain is expanded through the above-mentioned seed medium, carry out liquid submerged fermentation in the above-mentioned liquid fermentation medium in a fermenter at 28-35° C., accumulate antibacterial metabolites for 30-48 hours, and remove the bacteria through centrifugation of the fermentation liquid. Then the supernatant is concentrated, extracted with 3-5 times of methanol, and dried to obtain the finished antimicrobial peptide. The content of the antimicrobial peptide is not less than 500g / kg as determined by high performance liquid chromatography.

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Abstract

The invention relates to a composite antibacterial peptide produced by bacillus natto NT-6 and a production method thereof. The composite antibacterial peptide product is a mixture of three kinds of homologs of antibacterial lipopeptide substances iturins, fengycins and surfactins in the proportions of 10-20 percent, 25-35 percent and 50-60 percent respectively; the antibacterial peptide pure product is a yellow viscous substance which can resist heating of 30 minutes at the temperature of 100 DEG C, can keep activity at the pH of 2-12, has broad-spectrum antibacterial activity for bacteria and fungi and has strong killing effect on protozoa and cancer cells; and experiments prove that the peptide has high safety. The invention also discloses the production method of the composite antibacterial peptide. The peptide can be easily produced by using the process, is relatively low in cost, and can be used as a food preservative, a feed additive, an agricultural biological control agent, a medicament, an animal disease preventing agent and the like.

Description

technical field [0001] The invention relates to a compound antibacterial peptide produced by Bacillus natto NT-6 strain and a method thereof, belonging to the field of biotechnology. The antibacterial lipopeptide produced by the method has broad-spectrum antibacterial activity and biological surface activity, and has broad application prospects in the fields of medicine, agriculture, food and environmental governance. Background technique [0002] Bacterial tempeh (called natto in Japan) is a very common flavor-rich fermented soybean food in China with a history of more than 2,000 years. Its fermentation strain is Bacillus natto ( Bacillus natto). Bacillus natto can produce many functional factors, so that fermented products have many beneficial functions for the human body. At present, domestic and foreign research on Bacillus natto mainly focuses on the metabolite nattokinase with thrombolytic effect. And its antibacterial effect has only been paid attention to by peo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12P21/00A23L3/3526A23K1/17A01N63/02A01P1/00A01P3/00A61K38/02A61P31/00C12R1/07A23K20/195
Inventor 孙力军王雅玲刘唤明徐德峰聂芳红
Owner GUANGDONG OCEAN UNIVERSITY
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