Bacillus natto and its application in the preparation of ace-inhibiting peptides from fermented scallop skirts

A technology of scallop skirt and inhibitory peptides, applied in fermentation, application, bacteria and other directions, can solve the problem of reversible reduction of renal function, and achieve the effects of no side effect activity, high inhibitory activity, and simple operation.

Inactive Publication Date: 2021-02-23
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, drug treatment of high blood pressure often produces side effects such as coughing, decreased renal function reversibility, and skin rashes, so people are increasingly favoring food therapy

Method used

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  • Bacillus natto and its application in the preparation of ace-inhibiting peptides from fermented scallop skirts
  • Bacillus natto and its application in the preparation of ace-inhibiting peptides from fermented scallop skirts
  • Bacillus natto and its application in the preparation of ace-inhibiting peptides from fermented scallop skirts

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The separation and purification of embodiment 1 bacillus natto

[0040] Weigh 0.2g natto and shake it in 20mL 0.9% normal saline for 1h, and use this bacterial suspension as the original bacterial suspension; take 0.5mL of the original bacterial suspension and add it into 4.5mL sterile normal saline to make 10 -1 Dilute the bacterial suspension, prepare 10 by gradient dilution as above -2 ~10 -8 Dilute the bacterial suspension; each take 200μl of the above 10 -5 ~10 -8Add the diluted bacterial suspension to the beef extract peptone medium plate with a salt content of 2%, set 3 parallel plates for each treatment, and spread the dilution evenly; move the above plate to a 37°C constant temperature incubator and incubate it upside down for 24 hours; observe the growth of the colony , select a plate with uniform distribution of colonies and the number of colonies between 30 and 80, pick single colonies of different colony forms for Gram staining, and observe under a micro...

Embodiment 2

[0042] The preliminary screening of embodiment 2 bacillus natto

[0043] The 35 strains of Bacillus natto isolated above were planted on the solid medium of skim milk with a sterile toothpick, cultured upside down at 37°C for 18 hours, the diameter of the colony and its surrounding hydrolysis circle was measured, and the protease activity was roughly calculated.

[0044] Seven strains of Bacillus natto with large hydrolysis circle diameter were used to prepare crude enzyme solution, and the protease activity of the crude enzyme solution was determined by Folin-phenol method. Definition of protease activity: under the conditions of 40°C and pH 7.2, the number of micrograms of tyrosine produced per milliliter of fermentation broth (unit: IU / mL).

[0045] Prepare tyrosine standard solutions of 0, 20, 40, 60, 80, and 100 μg / mL, take 1 mL of the above standard solutions, add 5 mL of 0.4 mol / L sodium carbonate solution, and then add 1 mL of Folinol reagent, Vortex and oscillate to ...

Embodiment 3

[0052] The double screening of embodiment 3 bacillus natto

[0053] The above-mentioned 7 strains of Bacillus natto with high protease activity were activated and fermented the scallop skirt respectively, and the fermentation supernatant was taken, and the polypeptide content and ACE inhibition rate were used as indicators for re-screening.

[0054] Determination of polypeptide content in scallop skirt fermentation broth by biuret method. Prepare 0.0, 0.2, 0.4, 0.6, 0.8, 1.0 and 1.2 mg / mL Gly-Gly-Tyr-Arg tetrapeptide standard solutions in sequence with 5% trichloroacetic acid (TAC), then take 3.0 mL of the above standard solutions, Add 2.0mL biuret reagent, mix evenly on a vortex mixer, let it stand for 5min, take the supernatant and measure the absorbance value at 540nm (the first tube is used as the blank control), take the peptide concentration as the abscissa, OD 540 The value is the ordinate, making the Gly-Gly-Tyr-Arg tetrapeptide solution standard curve, such as imag...

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Abstract

The invention discloses the application of Bacillus subtilis subsp.natto ACE-IP and Bacillus subtilis subsp.natto ACE-IP in the preparation of ACE-inhibiting peptides from fermented scallop skirts, and its preservation number is CCTCC NO: M 2018081. The screening and identification method of Bacillus subtilis natto subspecies ACE‑IP includes: raw material processing and amino acid composition analysis; preparation of scallop skirt fermentation medium; primary screening of Bacillus natto; secondary screening of Bacillus natto; identification of strains . The Bacillus natto screened by the present invention is prepared by fermenting the scallop skirt to obtain the ACE inhibitory peptide, which fully exerts the effect of producing various proteases during the fermentation of the scallop skirt, and decomposes the scallop skirt macromolecular protein into small molecule ACE Inhibiting peptides with high inhibitory activity; not only provide excellent strains for industrial production of ACE inhibitory peptides that are safe, have no side effects and have high activity, but also provide a theoretical basis for the development, utilization and further processing of scallop skirts.

Description

technical field [0001] The invention relates to the field of functional food processing, in particular to a strain of natto bacteria and its application in preparing ACE inhibitory peptides from fermented clam scallop skirt. Background technique [0002] Chlamysfarreri belongs to the order Pterioida, Pteriina, Pectinidae, and the genus Chlamys. It is widely distributed in the coastal areas of northern China, such as Lvda in Liaoning and Shandong. Qingdao, Changdao, Rongcheng, Rizhao and other places. The scallop skirt is the remaining part after removing the scallop shell, which accounts for about 30% of the whole scallop. The output of Shandong Province alone is as high as 300,000 tons per year, but it is directly discarded due to seasonal restrictions in the production process. Reasonable and effective application has caused waste of resources and pollution of the environment. Studies have shown that scallop skirt contains a lot of protein, fat, carbohydrates and other n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P21/00A23L33/18C12R1/07
CPCA23V2002/00A23L33/18C12P21/00C12N1/205C12R2001/07A23V2200/326A23V2250/55
Inventor 魏玉西高洁高翔
Owner QINGDAO UNIV
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