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Bacillus subtilis, application thereof, and enzyme preparation

A technology of Bacillus subtilis and enzyme preparations, applied in the direction of application, enzymes, bacteria, etc., to achieve the effects of shortening the process flow, assisting the treatment of high blood pressure, and stabilizing production quality

Active Publication Date: 2018-12-07
SOUTH CHINA UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no enzyme preparation specially used for hydrolyzing yeast to prepare antihypertensive active polypeptides

Method used

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  • Bacillus subtilis, application thereof, and enzyme preparation
  • Bacillus subtilis, application thereof, and enzyme preparation
  • Bacillus subtilis, application thereof, and enzyme preparation

Examples

Experimental program
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Effect test

Embodiment 1

[0033] 1. Acquisition and identification of strain HU528

[0034] Soil was collected from a traditional Tibetan ghee workshop, 10g of protein-rich soil sample was added to 100mL of sterile water, shaken at 28°C for 30min to prepare a bacterial suspension, and the bacterial suspension was diluted and spread on a plate at 28°C Culture on solid medium (0.4% (w / w, the same below) beef extract, 0.6% tryptone, 0.2% yeast extract, 0.5% NaCl, 2% agar, adjust pH 7.0) for 1-2d. Inoculate single colonies with different growth traits obtained on the plate into LB medium (1% tryptone, 0.5% yeast extract, 1% NaCl, adjust pH 7.0) and cultivate for 48 hours, measure the protease activity in the fermentation broth, and obtain protease-producing strains , named strain HU528.

[0035] After the HU528 strain was streak cultured on a solid plate medium at 37°C for 24 hours, the colony was white and round, with rough and wrinkled surfaces and irregular edges ( figure 1 ). Microscopic observation...

Embodiment 2

[0053] Prepare the liquid medium as follows: add tryptone 2% (w / w, the same below), glucose 0.1%, CaCl in distilled water 2 0.2%, NaCl 0.1%, after mixing evenly, adjust the pH of the liquid medium to 5.0. After sterilizing at 121° C. for 20 minutes, the primary seeds of Bacillus subtilis HU528 were inoculated in the liquid medium at 1.0% (v / v), and the temperature was 30° C., and the stirring speed was 150 r / min. After fermentation and cultivation for 45 hours, The obtained fermentation liquid was centrifuged at 4° C. and 8000 r / m for 10 minutes to remove bacteria, and the fermentation supernatant was the enzyme preparation. The protease activity in the enzyme preparation was 800U / mL, and the β-glucanase activity was 2.0U / mL.

[0054] Add the above-mentioned enzyme preparation containing 2000U of protease activity (2.5mL enzyme preparation / g dry yeast) to the edible yeast powder by every gram of yeast powder, supplement sterile water to the volume ratio of edible yeast powder...

Embodiment 3

[0059] Prepare the liquid medium as follows: add tryptone 1.5% (w / w, the same below), glucose 0.8%, CaCl in distilled water 2 0.05%, NaCl 0.15%, after mixing evenly, adjust the pH of the liquid medium to 6.0. After sterilizing at 121°C for 20 minutes, inoculate the primary seeds of Bacillus subtilis HU528 in liquid medium at 1.0% (v / v), ferment and cultivate for 60 hours at a temperature of 25°C and a stirring speed of 180r / min. , the obtained fermentation broth was centrifuged at 4°C and 8000r / m for 10min to remove bacteria, and the fermentation supernatant was the enzyme preparation. The protease activity in the enzyme preparation was 1600U / mL, and the β-glucanase activity was 3.0U / mL.

[0060] Add the above-mentioned enzyme preparation containing 9600U of protease activity (6mL enzyme preparation / g dry yeast) to the edible yeast powder per gram of yeast powder, and add sterile water to the volume ratio of the edible yeast powder to the mixed liquid (solid-liquid ratio, w / v...

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Abstract

The invention discloses a type of Bacillus subtilis, which is named HU 528, is preserved in Guangdong Culture Collection Center at Apr 23rd, 2018, and is assigned the accession number of GDMCC No.60364. The invention also discloses an enzyme preparation prepared from the Bacillus subtilis, and also discloses an application of preparing a yeast enzymolysis product, having blood pressure reducing activity, by the Bacillus subtilis. The enzyme preparation prepared from the Bacillus subtilis HU 528 has the activities of protease and beta-glucanase and can destruct yeast cell wall and hydrolyze yeast protein. The yeast enzymolysis product has excellent bioactivity; in-vivo and in-vitro activity tests prove that the enzymolysis product at least has blood pressure reducing and anti-oxidizing activity and can be used as a non-medicinal treatment food for hypertension patients. The product can effectively prevent, control, alleviate and auxiliarily treat the hypertension.

Description

technical field [0001] The invention relates to the field of food biotechnology, in particular to a bacillus subtilis and its application and an enzyme preparation. Background technique [0002] Non-drug treatment is an important means to treat and control hypertension. The antihypertensive polypeptide products developed with yeast as raw materials are expected to become safe and green functional foods for non-drug treatment of hypertension. Yeast powder was hydrolyzed with commercially available yeast extract compound enzyme, and the polypeptide obtained after removing sugar and nucleic acid substances had ACE inhibitory activity; after further application of ultrafiltration membrane separation and column chromatography, a hexapeptide with high blood pressure lowering activity was obtained . Mirzaei, a foreign scholar, prepared ACE inhibitory peptides by using ultrasonic wall breaking to assist trypsin hydrolysis of yeast. [0003] Since the yeast cell wall is thick, it i...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/42C12N9/54C12N1/06A23L33/14C12R1/125
CPCA23V2002/00A23L33/14C12N1/06C12N9/244C12N9/54C12Y302/01006C12N1/205C12R2001/125A23V2200/326A23V2250/76
Inventor 胡松青王佳佳侯轶
Owner SOUTH CHINA UNIV OF TECH
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