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Cladosporium Cladosporium with High Ferulic Esterase Production and Its Application in Vinegar Brewing

A technology of cladosporium and vinegar, which is applied in the preparation of hydrolytic enzymes, fungi, vinegar, etc., can solve the problems of low ferulic acid esterase activity and the inability to realize industrial application, achieve bright color, improve ferulic acid The content of ferulic acid and the unique effect of refreshing taste

Active Publication Date: 2018-05-25
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is: aiming at the problem that the activity of microbial ferulic acid esterase is low at present and cannot realize industrial application, provide a new strain of Cladosporium cladosporium with high yield of ferulic acid esterase, which can utilize wheat Ferulic esterase is produced by fermentation of bran, which is added to the fermentation process of vinegar for the first time. The ferulic acid-rich vinegar produced has anti-oxidation, lowering blood pressure, anti-thrombotic, lowering blood fat, anti-bacterial and anti-inflammatory properties, cancer prevention, and health care. Liver, treatment of diabetes and other effects

Method used

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  • Cladosporium Cladosporium with High Ferulic Esterase Production and Its Application in Vinegar Brewing
  • Cladosporium Cladosporium with High Ferulic Esterase Production and Its Application in Vinegar Brewing
  • Cladosporium Cladosporium with High Ferulic Esterase Production and Its Application in Vinegar Brewing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Screening of Cladosporium strains

[0033] In the preliminary screening of Cladosporium cladosporium, the strains with the ability to produce ferulic acid esterase were selected by using ethyl ferulate as the only carbon source to produce a transparent circle phenomenon.

[0034] Take a sample of vinegar malt from Zhejiang Tapai Shaoxing Wine Co., Ltd., take 10g of malt, place it in a 250mL Erlenmeyer flask containing 90ml of sterile distilled water and glass beads, seal, shake on a shaker at 200r / min for 30min, make it Microorganisms are fully dispersed and make 10 -1 Times the sample dilution.

[0035] Use the gradient dilution method on the clean bench to get 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 Times the sample dilution, take 10 -4 , 10 -5 , 10 -6 , 10 -7 0.2 mL of each of the four dilution gradient samples were spread on the primary sieving medium, placed in an incubator at 28°C for 3 days, and then the size of the transparent circle was observed.

[00...

Embodiment 2

[0038] Example 2: Microbiological characteristics and identification of strains

[0039] The results of the strain after separation and purification on the PDA plate are as follows figure 2 As shown, the bacterial colony of the strain is thick, thick and fluffy, tightly bound to the culture medium, and not easy to provoke. The color ranges from smoky brown to greenish brown at first, and dark brown to black at the later stage. The back of the colony is black. Initially judged to be mold.

[0040] The total DNA of the strain was extracted as a template, and the rDNAITS1-5.8S-ITS2 region was amplified by PCR. The sequence length of the target gene was 520bp after sequencing, and the GenBank sequence number: KP900248.1. The identity of the rDNAITS1-5.8S-ITS2 sequence of the strain with the 10 strains is above 90.0%, and the identity with the Cladosporium cladosporioides strain FR5-SAR1 reaches 100%. The molecular evolutionary tree is constructed, and you can see that the strain S2...

Embodiment 3

[0041] Example 3: Application of strain wheat bran fermentation broth in vinegar production

[0042] Wheat bran 0.5kg, distilled water 10L, pH4.8, oxygen 1vvm, rotation speed 150r / min, temperature 28℃, fermentation was carried out, the liquid fermentation broth at the 76th hour of fermentation was taken, and ferulic acid esterase was measured by high performance liquid chromatography Activity (e.g. image 3 Shown), the enzyme activity is 280U / L.

[0043] Chromatographic column: XBridgeTM C18 (4.6×250mm, 5μm)

[0044] Mobile phase: (B: 0.01% glacial acetic acid aqueous solution; mobile phase D: pure acetonitrile). The flow rate is 1.0 mL / min.

[0045] Gradient elution method: start time, B solution flow rate 80%, D solution flow rate 20%, maintain 2min; 8min, B solution flow rate 72%, D solution flow rate 28%; 13min, B solution flow rate 42%, D solution flow rate 58 %; 19min, the flow rate of B solution is 80%, the flow rate of D solution is 20%, maintain for 6min, end. The linear ...

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Abstract

The invention discloses a strain of cladosporium with high ferulic acid esterase production and its application in vinegar brewing, belonging to the fields of fermentation engineering and biotechnology. The Cladosporium cladosporioides of the present invention is preserved by the China Center for Type Culture Collection, the preservation number is CCTCC NO: M 2015549, and the preservation date is September 16, 2015. The invention uses wheat bran as a fermentation culture substrate to ferment and produce highly active ferulic acid esterase, which is easy to realize the industrialized production of ferulic acid esterase. And ferulic acid esterase was applied in the production of vinegar, and the functional health-care vinegar rich in ferulic acid was obtained, which provided new technical support for the development of health-care vinegar.

Description

Technical field [0001] The invention relates to a cladosporium strain with high production of ferulic acid esterase and its application in vinegar brewing, belonging to the fields of fermentation engineering and biotechnology. Background technique [0002] Ferulic acid esterase (FEA, EC 3.1.1.73) is a subclass of carboxylate hydrolases. Its main biological function is to hydrolyze the ester bond between plant cell wall polysaccharides and ferulic acid (FA), thereby combining ferulic acid with Other phenylacrylic acid is released. At present, there are about 30 strains reported to produce ferulic acid esterase, which mainly include Aspergillus niger, Aspergillus oryzae, Aspergillus terreus, Aspergillus flavus, Aspergillus flavus, Aspergillus awamori, Aspergillus tubing, Aspergillus nidulans, Aspergillus usami, Rhizopus, Penicillium, Pleurotus eryngii, Fusarium oxysporum, Fusarium spp., Neurospora crassa, Rhizomucor miehei, Basiliforme, Russula, Pleurotus eryngii, Aureobasidium pu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/18C12J1/04C12R1/645
Inventor 毛健李翠翠刘双平姬中伟孟祥勇
Owner JIANGNAN UNIV
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