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Moringa seed protease capable of promoting goat milk solidification and polypeptide prepared from moringa seed protease and application

A technology of Moringa oleifera seeds and protease, applied in the field of dairy science, can solve the problems of unpredictable polypeptide functional activity, inability to guarantee added value, few reports on the research of protease curd function, etc., to achieve the effect of enhancing functional activity

Active Publication Date: 2020-04-14
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Milk coagulation is a protease that specifically cleaves the peptide bond of κ-casein in milk and destroys the casein micelles. In this process, a large number of polypeptide fragments will be produced. However, the cleavage site of each protease to cleave κ-casein is different. , the polypeptides produced by cleavage are not the same, and whether the produced polypeptides have functional activity cannot be predicted, so the added value of cheese, milk cake, yogurt and other products after protease curdling cannot be guaranteed
[0004] Moringa tree is a perennial tropical and subtropical deciduous tree, belonging to the genus Moringa of Moringaceae Moringa. Functional studies have rarely been reported

Method used

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  • Moringa seed protease capable of promoting goat milk solidification and polypeptide prepared from moringa seed protease and application
  • Moringa seed protease capable of promoting goat milk solidification and polypeptide prepared from moringa seed protease and application
  • Moringa seed protease capable of promoting goat milk solidification and polypeptide prepared from moringa seed protease and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The extraction of embodiment 1 Moringa seed protease

[0033] 1) Get 100 grams of Moringa seeds (Indian species PKm2, tree age 3 years), the Moringa seeds moisture content is 10.5%, the Moringa seeds are obtained through mechanical pulverization to obtain the Moringa seeds powder, then extract the Moringa seeds by supercritical carbon dioxide The powder is degreased. The conditions for degreasing treatment of Moringa oleifera seed powder by supercritical carbon dioxide extraction are: under the conditions of extraction pressure 30MPa, extraction temperature 40°C, and separation temperature 40°C, 26.7% of the fat in Moringa oleifera seeds is removed, and the degreasing rate reaches 72.8%.

[0034] 2) Extraction of Moringa seed crude protein: use the salt method to process the defatted Moringa seed powder to obtain the crude protein extract of Moringa seed; the optimal extraction process parameters are solid-liquid ratio 1:10, extraction temperature 30 ℃, pH=7.15, salt c...

Embodiment 2

[0047] The identification of embodiment 2 Moringa seed protease

[0048] 1) Reductive alkylation treatment (10mM TCEP[Tris(2-carboxyethyl)phosphine] at 37°C for 60 minutes, 40mM iodoacetamide at room temperature for 40 minutes) was performed on the purified Moringa seed protease, followed by enzymatic After treatment (trypsin, 18h), peptides were extracted for LC-MS / MS. Use the PEAKS Studio 8.5 software to retrieve data from the Moringa oleifera transcriptome database to identify the Moringa oleifera seed protease, and the identification results are as follows figure 2 As shown in the figure, the ion selection device automatically selects peptide ions with higher intensity to simulate fragmentation (secondary fragment ion spectrum matching)-mass spectrum prediction scoring evaluation. The peptide score will be calculated based on the size of the fragmented ions in the computer simulation and the matching degree in the actual secondary spectrum, and the one with the highest m...

Embodiment 3

[0053] Example 3 The cleavage site of κ-casein digested by Moringa oleifera seed protease and the polypeptide produced by enzymatic digestion

[0054] 1) Use the purified Moringa oleifera protease and κ-casein (κ-CN) at a ratio of 1:10 to react in a water bath at 60°C for 60 minutes, and then centrifuge the obtained protein solution through a Millipore ultrafiltration centrifuge tube (molecular weight 3k Da) at 4°C After desalting and removing impurities, the protein gel strips in which Moringa oleifera seed protease interacted with κ-casein for 60 minutes were obtained by SDS-PAGE, and the gel was cut for matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF / TOF), and the molecular weight, sequence and restriction sites.

[0055] The amino acid sequence of κ-casein is as follows. The amino acid sequence information of κ-casein comes from NCBI database, and the species source is bovine:

[0056]

[0057]

[0058] The digestion site is G...

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Abstract

The invention provides moringa seed protease capable of promoting goat milk solidification and polypeptide prepared from the moringa seed protease and an application of the moringa seed protease. Theenzyme digestion site of the protease enzyme digestion kappa-casein is Gln 113 to Asp 114. Goat milk can be solidified, and high milk coagulation activity is achieved. More importantly, the moringa seed protease performs enzyme digestion on kappa-casein to obtain four protein peptides with better antioxidant activity. The moringa seed protease disclosed by the invention is used as a curding agentfor curding processing of milk cakes, cheese and yoghourt, and the obtained product is rich in antioxidant peptides. Compared with the prior art, the moringa seed protease can enhance the functional activity of goat milk and the product of goat milk.

Description

technical field [0001] The invention belongs to the field of dairy science and technology, and in particular relates to a Moringa seed protease capable of promoting coagulation of goat milk, a polypeptide prepared therefrom and applications thereof. Background technique [0002] Milk coagulation is a necessary link in the production of cheese, milk cake, and yogurt. Coagulants play an important role in product development and product quality improvement. The rapid development of the food industry (especially the cheese industry) has led to a shortage of coagulants, and vigorously develop new resources. It must be done. [0003] Milk coagulation is a protease that specifically cleaves the peptide bond of κ-casein in milk and destroys the casein micelles. During this process, a large number of polypeptide fragments will be produced. However, the cleavage site of each protease to cleave κ-casein is different. , The polypeptides produced by cleavage are also different, and whet...

Claims

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Application Information

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IPC IPC(8): A23C19/04A23C9/12C12N9/50C07K14/47C12P21/06
CPCA23C9/1209A23C19/041C07K14/4732C12N9/63C12P21/06
Inventor 黄艾祥施娅楠王雪峰赵存朝
Owner YUNNAN AGRICULTURAL UNIVERSITY
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