Bitterness-free shrimp-type low-molecular-weight peptides as well as preparation method and application thereof

A low-molecular-weight peptide, no bitterness technology, applied in the field of shrimp low-molecular-weight peptides and their preparation, can solve the problems that the enzymatic hydrolysis degree of low-molecular-weight peptides is difficult to control, the yield of specific products is low, and the production cost is high, and the bitterness removal effect is good. , The effect of saving separation and purification costs and saving energy costs

Active Publication Date: 2016-11-16
GUANGDONG IND TECHN COLLEGE +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the flavor protease preparation is a mixed preparation of endo-protease and exo-protease. When it is used for enzymatic hydrolysis and debittering, the degree of enzymatic hydrolysis of low-molecular-weight peptides is difficult to control, and it is easy to cause the product molecule to be too small and the yield of specific products to be too low.
In addition, the cost of aminopeptidase extraction and purification accounts for about 70% of the production cost of enzyme preparations. If aminopeptidase preparations are used directly for debittering, there is also the disadvantage of high production costs

Method used

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  • Bitterness-free shrimp-type low-molecular-weight peptides as well as preparation method and application thereof
  • Bitterness-free shrimp-type low-molecular-weight peptides as well as preparation method and application thereof
  • Bitterness-free shrimp-type low-molecular-weight peptides as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] (1) Isolation and screening of high aminopeptidase-producing strains

[0052] Enrichment medium: glucose 50g / L, peptone 10g / L, KH 2 PO 4 2g / L, MgSO 4 ·7H 2 O0.5g / L, NaCl5g / L, pH7.0, sterilize at 121°C for 20min, cool for later use.

[0053] Plate separation medium: glucose 5g / L, casein 15g / L, KH 2 PO 4 1g / L, NaCl 5g / L, agar 20g / L, pH 7.0, sterilize at 121°C for 20min, cool for later use.

[0054] Slant medium: glucose 5g / L, peptone 5g / L, yeast extract 5g / L, NaCl 5g / L, agar 20g / L, pH7.0, sterilized at 121°C for 20min, cooled for later use.

[0055] Fermentation medium: soluble starch 40g / L, peptone 20g / L, KH 2 PO 4 1.5g / L, MgSO 4 ·7H 2 O 0.5g / L, pH7.0, sterilized at 121°C for 20min, cooled for later use.

[0056] Douchi is a commercially available material. 10 g of Douchi was added to 200 mL of enrichment medium, and cultured with shaking at 30° C. and 200 rpm for 16 hours. Dilute the culture solution in gradient, spread it on the plate separation medium, ...

Embodiment 2

[0067] (1) Shrimp enzymatic hydrolysis

[0068] Grind the dried shrimp through a 100-mesh sieve to obtain a powder; weigh 1200g of the powder (56.2% protein content), add 18L of water, and mix well; then add 56.2g of neutral protease, and enzymatically hydrolyze at 55°C for 4h , and then heated to 85°C for 12 minutes to inactivate the enzyme, and cooled to room temperature to obtain the enzymatic hydrolysis solution;

[0069] (2) Preparation of seed solution

[0070] Take 900mL of enzymolysis liquid, add 18g of soluble starch and 1.8g of potassium dihydrogen phosphate, adjust the pH to 7.0, and put it into five 1000mL Erlenmeyer flasks evenly, after sterilization and cooling, insert Bacillus subtilis YBPE-4 (CCTCC NO: M 2016359) slant strains, each bottle was inserted with 2 rings of bacterial lawn, cultivated at 37°C and 160rpm for 24h, and the bacterial concentration was 1.08×10 10 Individual / mL seed solution;

[0071] (3) Fermentation debittering of low molecular weight ...

Embodiment 3

[0076] (1) Shrimp enzymatic hydrolysis

[0077] Grind the dried shrimp through an 80-mesh sieve to obtain a powder; weigh 900g of the powder (56.2% protein content), add 18L of water, and mix well; then add 33.72g of neutral protease, and enzymatically hydrolyze at 50°C for 5h , then heated to 80°C, kept at a constant temperature for 15 minutes, inactivated the enzyme, cooled to room temperature, and obtained the enzymatic hydrolysis solution;

[0078] (2) Preparation of seed solution

[0079] Take 1800mL of enzymolysis solution, add 27g of soluble starch and 2.7g of potassium dihydrogen phosphate, adjust the pH to 6.5, and put it into ten 1000mL conical flasks evenly, after sterilization and cooling, insert Bacillus subtilis YBPE-4 (CCTCC NO: M 2016359) slant strains, each bottle was inserted with 2 rings of bacterial lawn, cultivated at 33°C and 200rpm for 28h, and the concentration of bacteria obtained was 2.49×10 10 Individual / mL seed solution;

[0080] (3) Fermentation...

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Abstract

The invention discloses a bitterness-free shrimp-type low-molecular-weight peptides as well as a preparation method and application thereof, belonging to the technical field of biology. The invention provides bacillus subtilis YBPE-4 with a preservation number of CCTCC NO: M2016359. The preparation method comprises the following steps: drying and crushing shrimps, enzymolyzing the shrimps by using neutral protease, and separating to obtain an enzymolysis solution; preparing a fermentation culture medium by utilizing the enzymolysis solution, soluble starch and potassium dihydrogen phosphate, inoculating a mature bacillus subtiliss YBPE-4 culture solution, and stirring, ventilating and fermenting for 132 to 144 hours; centrifuging a fermented solution to remove thallus to obtain supernatant, and filtering by virtue of an ultrafiltration membrane to obtain filtrate with a molecular weight of 360 to 5000 Da; and drying the filtrate to finally obtain the bitterness-free shrimp-type low-molecular-weight peptide product. The bitterness-free shrimp-type low-molecular-weight peptides have the advantages that the bitterness removal effect is good and the yield of products is high, and the preparation method is relatively low in production cost, suitable for producing the bitterness-free low-molecular-weight peptides by utilizing low-value shrimps in an industrialization manner and good in application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a non-bitter shrimp low-molecular-weight peptide and its preparation method and application. Background technique [0002] my country is a big country in aquaculture and processing. In recent years, the annual output of shrimp alone has exceeded 1 million tons. Among them, shrimp processing produces a large amount of leftovers, and a large number of small shrimps that are difficult to process are obtained from fishing, accounting for more than 40% of the total amount of shrimp, which can only be used to produce animal feed or shrimp paste food. Shrimp processing waste and low-value shrimp are rich in protein, and have recently become important resources for people to develop products such as amino acids, active peptides, and seasoning bases. Low-molecular-weight peptides have the characteristics of easy absorption and taste, and their different flavors depend on the leng...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00C12P21/06C07K1/34A23L33/18C12R1/125
CPCC07K1/34C12P21/00C12P21/06C12N1/205C12R2001/125
Inventor 李静邓毛程张东峰王瑶陆志鸿邱燕翔叶茂叶枫周慧娉陈光
Owner GUANGDONG IND TECHN COLLEGE
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