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A method for co-producing yeast glucan, mannoprotein and yeast extract using β-1,6 glucanase

A technology of yeast extract and yeast glucan, applied in the field of co-production of yeast glucan, mannoprotein and yeast extract, can solve the problems of unknown enzymatic activity and cost, and avoid the complexity of equipment and operation. , reduce cost, improve the effect of purity

Active Publication Date: 2022-08-09
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in CN1940084A, three biological enzymes that can break the overall structural connection of yeast cell walls are used, and their enzymatic activity and cost are unknown

Method used

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  • A method for co-producing yeast glucan, mannoprotein and yeast extract using β-1,6 glucanase
  • A method for co-producing yeast glucan, mannoprotein and yeast extract using β-1,6 glucanase
  • A method for co-producing yeast glucan, mannoprotein and yeast extract using β-1,6 glucanase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 A method for co-producing yeast glucan, mannoprotein and yeast extract using β-1,6 glucanase, using baker's yeast as a raw material, comprising the following steps (such as figure 1 shown):

[0054] (1) Using baker's yeast as a raw material, the yeast cells are prepared into a suspension with a mass percentage concentration of 9%, and the temperature is 110 ° C, the time is 20 minutes, and the pressure is 0.1 Mpa. Under the conditions of high temperature inactivation, the suspension The liquid was centrifuged at 8000rpm for 10min, and the precipitate and the supernatant were collected respectively: the precipitate was the treated yeast cell wall; the supernatant was concentrated and spray-dried to obtain the yeast extract;

[0055] (2) Corallococcus sp. EGB was inoculated in VY / 4 liquid enzyme production medium (0.40% yeast cell wall, 0.1% CaCl 2 , 0.1% yeast extract (w / v), pH 7.0), fermented and cultured for 2 days, collected the enzyme liquid by centrifugat...

Embodiment 2

[0066] About 20 mg of yeast glucan and mannoprotein obtained in Example 1 were weighed, 10 mL of trifluoroacetic acid (4M) was added, and hydrolyzed at 100° C. for 4 h. The hydrolyzate was rotary evaporated at 80°C and adjusted to neutrality with 4M NaOH. Pipette 150 μL of the hydrolyzate and add 150 μL of 0.6 M NaOH, then add 200 μL of 0.5 M PMP (1-phenyl-3-methyl-5-pyrazolone) methanol solution, and react in a water bath at 70°C for 90 min. After the reaction was completed, it was cooled to room temperature, 0.3 M HCl was added to the reaction solution to adjust to neutrality, and deionized water was added to 1 mL. Add 1 mL of chloroform to extract PMP, centrifuge at 12 000 rpm for 1 min, and repeat the extraction three times. The upper aqueous phase solution was passed through a 0.22 μm aqueous phase filter, and 20 μL was taken on the machine. The derivatized samples were chromatographed on a C18 column with UV detector OD 230 , the mobile phase was a mixed solution of P...

Embodiment 3

[0069] The content of each component of yeast glucan and mannoprotein obtained in Example 1 was determined. Crude protein and ash were measured by AOAC method (1990); total sugar was measured by anthrone-sulfuric acid method; yeast glucan and mannoprotein were measured by referring to Example 2, and quantified by external standard method.

[0070] The results are shown in Table 1. The yeast cells are mainly composed of polysaccharides and proteins, with a total sugar content of 34.32% (wherein, glucan accounts for 20.56% of dry cell weight, and mannan accounts for 13.32%), and protein content is 59.31%. The yield of mannoprotein prepared by the method of the invention is 11.1% and the purity is 90.66%; the yield of yeast glucan is 17.44% and the purity is 85.36%.

[0071] Table 1 Analysis of yeast and zymosan components

[0072]

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Abstract

The invention discloses a method for co-producing yeast glucan, mannoprotein and yeast extract by utilizing β-1,6 glucanase. The method uses yeast cells as raw materials and inactivates the yeast cells at high temperature, Centrifuge, dry the supernatant to obtain yeast extract; use β-1,6 glucanase to enzymatically hydrolyze the yeast cell wall, centrifuge and centrifuge to obtain the enzymatic hydrolysis supernatant and the enzymatic precipitation respectively, and the enzymatic hydrolysis supernatant is subjected to water extraction and alcohol precipitation , centrifugal separation, and drying to obtain mannoprotein; after ethanol is recovered, the remaining liquid part is concentrated and spray-dried to obtain yeast extract; the enzymatic precipitation is degreasing, protease treatment, and spray-drying to obtain yeast glucan. The yeast glucan and mannoprotein prepared by the invention have high purity, high yield, and low cost in the preparation process, do not use strong acid and strong alkali reagents, and cause little environmental pollution.

Description

technical field [0001] The present invention relates to a method for co-producing yeast glucan, mannoprotein and yeast extract by utilizing beta-1,6 glucanase. Background technique [0002] Yeast is an important food and industrial microorganism, and my country has abundant yeast resources. Yeast cell walls are composed of polysaccharides, proteins and a small amount of lipids and chitin. The two main polysaccharides are β-D-glucan, which accounts for about 50-55% of the dry weight of the cell wall, and is distributed on the inner side of the cell wall, forming the cross-linked skeleton structure of the cell wall; mannan proteins are distributed on the outer side of the cell wall. , about 40-45% of the dry weight of the cell wall, imparting biological activity to cells and controlling cell wall pore size. Both β-glucan and mannan proteins have rich biological functions. Yeast β-glucan has a unique triple helix structure, which can specifically bind to the body's immune ce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/395C07K1/14C07K1/34C08B37/02C12N1/06C12R1/865
CPCC07K14/395C12N1/06C07K1/34C07K1/14C12R2001/865C12N1/185
Inventor 崔中利乔燕叶现丰李周坤曹慧
Owner NANJING AGRICULTURAL UNIVERSITY
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