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Penaeus japonicus head ACE (angiotensin converting enzyme) inhibiting peptide and preparation method thereof

A bamboo shrimp and inhibitory peptide technology, which is applied in the biological field to achieve the effects of improving yield, strong ACE inhibitory activity, and improving hydrolysis rate

Inactive Publication Date: 2017-09-19
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no research report on the preparation of ACE inhibitory peptides using the heads of bamboo shrimps. However, the research on the ACE inhibitory activity of the heads of bamboo shrimps can not only make full use of the leftovers of bamboo shrimp heads, but also improve the activity of bamboo shrimps. Product added value, and has important theoretical significance for expanding research on ACE inhibitory peptides

Method used

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  • Penaeus japonicus head ACE (angiotensin converting enzyme) inhibiting peptide and preparation method thereof
  • Penaeus japonicus head ACE (angiotensin converting enzyme) inhibiting peptide and preparation method thereof
  • Penaeus japonicus head ACE (angiotensin converting enzyme) inhibiting peptide and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1: Preparation of ACE-inhibiting peptide from bamboo shrimp head

[0031] 1.1 Raw materials and reagents

[0032] Raw materials: bamboo shrimp head, purchased from Zhoushan Haisheng Biological Co., Ltd.

[0033] Reagents: trypsin, bromelain, pepsin, alkaline protease, neutral protease, alkaline protease and acid protease. B12, aprotinin, cytochrome C, bovine serum albumin, acetonitrile (chromatographic grade), trifluoroacetic acid (chromatographic grade), formic acid (chromatographic grade), chromatographic column packing Sephadex G-25 (Sigma, USA); anhydrous Ethanol, 2,6-di-tert-butyl-4-methylphenol (BHT), hydrochloric acid, sodium hydroxide and other reagents were of analytical grade (Sinopharm Chemical Reagent Co., Ltd.).

[0034] 1.2 Instruments and equipment

[0035] WATERS SYNAPT ultra-high pressure liquid chromatography quadrupole tandem time-of-flight mass spectrometer (Waters, USA); UPLC BEH C18 column (2.l ID×120mm) (Waters, USA); Sunfire TM C18 c...

Embodiment 2

[0090] Example 2: Identification of ACE inhibitory peptide structure

[0091] The ACE inhibitory component SHP-C after separation and purification by reverse-phase high-performance liquid phase was identified by UPLC-TOF-MS / MS technology, and its structure was analyzed by peptide sequencing in Masslynx software. The results showed that from SHP-C The components identified a polypeptide sequence with a reliability of more than 90%, and its structure was His-Ala-Phe (such as Figure 7 shown).

Embodiment 3

[0092] Embodiment 3: Determination of biological stability of ACE inhibitory peptide of bamboo shrimp head

[0093] Prepare artificial gastric juice and artificial intestinal juice for later use. The preparation process of artificial gastric juice is as follows: Measure 16.4mL of dilute hydrochloric acid and 800mL of distilled water respectively, mix them, then add 10g of pepsin, shake evenly, place in a volumetric flask, and constant volume to 1000mL. Preparation of artificial intestinal juice: Weigh 6.8g of potassium dihydrogen phosphate, dissolve it in 500mL of distilled water, adjust the pH value to 6.8 with 0.1mol / L NaOH solution, and take 10g of trypsin, dissolve it in a certain Amount of distilled water, shake evenly, and set the volume to 1000mL.

[0094] Prepare the same concentration (10mg / ml) of chicken ovalbumin solution and bamboo shrimp head ACE inhibitory peptide solution, mix them in the ratio of (1:4, 1:5, 1:6), and then add the same amount of prepared The s...

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Abstract

The invention relates to a penaeus japonicus head ACE (angiotensin converting enzyme) inhibiting peptide and a preparation method thereof. The amino acid sequence of the penaeus japonicus head ACE inhibiting peptide is His-Ala-Phe. The preparation method includes the steps: raw material pretreatment; ultrasonic-assisted enzymolysis; filter membrane ultrafiltration treatment; ion exchange column treatment; gel column treatment; reverse efficient liquid chromatography. The penaeus japonicus head ACE inhibiting peptide has strong ACE inhibiting activity, penaeus japonicus heads serve as raw materials, the problem of leftover generation in the processing process of penaeus japonicus is solved, and the penaeus japonicus head ACE inhibiting peptide has theoretical significance for further improving economic values of the penaeus japonicus. Besides, ultrasonic treatment is combined with protease enzymolysis, hydrolysis rate is increased, effective components in the raw materials are sufficiently extracted, extraction rate is increased, an extraction mode is mild, the effective components to be extracted cannot be damaged, and the penaeus japonicus head ACE inhibiting peptide with a reliable structure is obtained by multiple separation and purification and stepwise selection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an ACE-inhibiting peptide of the bamboo shrimp head and a preparation method thereof. Background technique [0002] Bamboo prawn (Penaeus japonicus) is scientifically known as Penaeus japonicus, which belongs to Decapod order and Penaeidae. It has the reputation of "Queen of Shrimp". Bamboo prawns are not only large in size, but also delicious in quality. They are high-quality seafood products, so they are very suitable for making sushi prawns and other products. [0003] At present, most bamboo shrimps are processed into headless quick-frozen shrimp meat products for domestic sale or export, so that about 35% of the shrimp head scraps are produced during the processing process, and the shrimp head meat accounts for about 10% of the shrimp head scraps. 30% by weight. Since the shrimp head contains hard sand and gravel components such as stomach pouches and mouthparts, it cannot be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/097C12P21/06C07K1/18C07K1/16C07K1/20C07K1/34
CPCC07K5/0821
Inventor 陈小娥冯淑莹方旭波李丽黄湛媛何定芬余辉
Owner ZHEJIANG OCEAN UNIV
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