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Method for preparing antihypertensive peptide from hairtail backbone by fermentation method

A blood pressure-lowering peptide and fermentation technology, applied in the field of bioengineering, can solve the problems of hairtail spine utilization and deep processing that are still blank, and achieve strong industrial implementation, low price, and the effect of inhibiting ACE activity

Inactive Publication Date: 2011-05-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is mainly used in the feed industry, and it is still blank for the utilization and deep processing of the hairtail backbone

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The culture conditions of Bacillus licheniformis prior to inoculation are:

[0036] Medium (g / L): beef extract, 4.0; peptone, 6.0; yeast extract, 2.0; NaCl, 5.0; pH, 7.5. Sterilized at 121° C. for 30 min.

[0037] Cultivation conditions: 250mL conical flasks are filled with 25mL seed medium, Bacillus subtilis (mycelium pre-preserved on the slant medium), placed on a rotating shaker at 37°C, 200rpm, and cultured for 16-18h to obtain an inoculum. The bacterial biomass (dry weight) in the inoculum is 0.4-0.5 g / L.

[0038] Preparation of blood pressure lowering peptides:

[0039] In a 250mL conical flask, weigh 4.5g of homogenized hairtail spine (moisture 76.22%, protein content 12.17%, provided by Ningbo Chaoxing Marine Biological Products Co., Ltd., Zhejiang Province), 1.2g of sucrose, 0.15g of peptone, K 2 HPO 4 , 0.03g, MgSO 4 ·7H 2 O, 0.003g, add 25mL of water, adjust pH to 7.5. Sterilize at 121°C for 30min.

[0040] Add 3% of the inoculum by volume (the inoculum...

Embodiment 2

[0041] Example 2 Preparation of blood pressure lowering peptides

[0042] In a 5L fermenter, weigh 600g of sucrose spine (water 76.22%, protein content 12.17%, provided by Ningbo Chaoxing Marine Biological Products Co., Ltd.), sucrose 160g, peptone 20g, K 2 HPO 4 , 4g, MgSO 4 ·7H 2 O, 0.4g, add 3400mL of water, adjust pH to 7.5. Sterilize at 121°C for 30min.

[0043] Add 3% of the inoculum by volume, 37 ° C, after stirring and culturing for 24 hours, to obtain a fermentation broth, the fermentation broth was centrifuged for 30 minutes at 4 ° C at 3000 r / min in a J-6M large-capacity refrigerated centrifuge (BECKMAN company), The supernatant was filtered to obtain 3.6kg of filtrate. The filtrate was placed in a 100°C water bath to rapidly heat up to about 90°C and kept for 10-15min to kill the enzyme. Then it was quickly cooled to room temperature, centrifuged at 3000 rpm for 15 min, and the supernatant was decolorized and deodorized by adding activated carbon at 25%, and f...

Embodiment 3

[0044] Example 3 Preparation of blood pressure lowering peptides

[0045]In a 5L fermenter, weigh 300g of the hairtail spine (water 76.22%, protein content 12.17%, provided by Ningbo Chaoxing Marine Biological Products Co., Ltd., Zhejiang Province), 160 g of glucose, K 2 HPO 4 , 4g, MgSO 4 ·7H 2 O, 0.4g, add 3600mL of water, adjust pH to 7.5. Sterilize at 121°C for 30min.

[0046] Add 3% of the inoculum by volume, 37 ° C, after stirring and culturing for 24 hours, to obtain a fermentation broth, the fermentation broth was centrifuged for 30 minutes at 4 ° C at 3000 r / min in a J-6M large-capacity refrigerated centrifuge (BECKMAN company), The supernatant was filtered to obtain 3.35kg of filtrate. The filtrate was placed in a 100°C water bath to rapidly heat up to about 90°C and kept for 10-15min to kill the enzyme. Then it was quickly cooled to room temperature, centrifuged at 3000 rpm for 15 min, and the supernatant was decolorized and deodorized by adding activated carbo...

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Abstract

The invention provides a method for preparing antihypertensive peptide by using ridge bones of hairtail in a fermentation way. The method comprises the following steps: (1), sugar, crushed hairtail ridge bone, peptone, K2HPO4, MgSO4.7H2O and water are mixed to produce fermentation medium, after sterilization, bacillus licheniformis is inoculated, and is cultured to obtain fermentation liquid; (2), after the fermentation liquid is centrifuged, the supernatant liquid is taken and filtered, and enzyme sterilization is performed to the filter liquor; (3), the filter liquor after the enzyme sterilization is centrifuged, discoloring and smell removal are performed to the centrifuged supernatant liquid by activated carbon, so as to obtain the filter liquor after the discoloring and the smell removal; (4), the filter liquor after the discoloring and the smell removal is ultrafiltered to obtain permeation liquid, freeze drying is performed to the permeation liquid after vacuum concentration, and then peptide powder with antihypertensive function is obtained. The sources of the raw material adopted by the method of the invention are rich, the price is low, the operation is easy, and the method has stronger industrialized practicability.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for preparing blood pressure-lowering peptides by using hairtail spines by a fermentation method. Background technique [0002] Angiotensin-converting enzyme (ACE) plays an important role in the regulation of blood pressure. It converts the inactive angiotensin decapeptide precursor to angiotensin, which increases blood pressure. Angiotensin-converting enzyme (ACE) inhibitory active peptide is a kind of angiotensin-converting enzyme inhibitor (ACEI), which can reduce blood pressure by inhibiting the activity of ACE. At present, a variety of chemically synthesized peptide drugs have been used as ACE inhibitors. The disadvantage of this kind of synthetic hypotensive drugs is that the action time is not long, the blood pressure is easy to rebound, and the drug is easily excreted by the kidneys, which can easily cause damage to renal function. [0003] Extracting nutritious ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00C12R1/10
Inventor 王金玲何国庆董晓美陈启和阮晖章海峰
Owner ZHEJIANG UNIV
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