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Cladosporiumc cladosporioides for high yield of feruloyl esterase and application of Cladosporiumc cladosporioides in edible vinegar brewing

A technology of Cladosporium cladosporium and ferulic acid esterase, which is applied in the directions of hydrolase, fungi, vinegar preparation, etc., can solve the problems of low ferulic acid esterase activity, unable to realize industrial application, etc., and achieve bright color and luster. , Increase the content of ferulic acid, refreshing and unique effect

Active Publication Date: 2016-01-13
JIANGNAN UNIV
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  • 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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  • Cladosporiumc cladosporioides for high yield of feruloyl esterase and application of Cladosporiumc cladosporioides in edible vinegar brewing
  • Cladosporiumc cladosporioides for high yield of feruloyl esterase and application of Cladosporiumc cladosporioides in edible vinegar brewing
  • Cladosporiumc cladosporioides for high yield of feruloyl esterase and application of Cladosporiumc cladosporioides in edible vinegar brewing

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Screening of Cladosporium strain

[0033] For the primary screening of Cladosporium cladoides, strains with the ability to produce ferulic acid esterase were screened out by using ethyl ferulate as the sole carbon source and producing a transparent circle phenomenon.

[0034] Take vinegar wheat koji samples from Zhejiang Tapai Shaoxing Wine Co., Ltd., take 10g of wheat koji, place it in a 250mL triangular flask containing 90ml sterile distilled water and glass beads, seal it, and shake it on an oscillator at 200r / min for 30min to make it Microorganisms are fully dispersed to make 10 -1 double sample diluent.

[0035] Obtain 10 by serial dilution method on ultra-clean bench -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 times sample diluent, take 10 -4 、10 -5 、10 -6 、10 -7 Spread 0.2mL of each of the four diluted gradient samples on the primary screening medium, place them in an incubator at 28°C for 3 days, and observe the size of the transparent circle....

Embodiment 2

[0038] Embodiment 2: Microbiological characteristics and identification of bacterial strains

[0039] The results after the strains were isolated and purified on the PDA plate were as follows: figure 2 As shown, the colony of the strain is thick, thick and villous, closely combined with the medium, and not easy to provoke. The color is smoky brown to greenish brown at the beginning, and dark brown to black later. The back of the colony is black. It was preliminarily 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 length of the sequenced target gene sequence was 520bp, GenBank sequence number: KP900248.1. The rDNAITS1-5.8S-ITS2 sequence of the strain has more than 90.0% identity (Identity) with the 10 strains, and has 100% identity with Cladosporium cladosporioidesstrain FR5-SAR1. Cladosporium cladosporioides is on the same branch of the constructed phylogenetic tree, combined ...

Embodiment 3

[0041] Embodiment 3: the application of bacterial strain wheat bran fermented liquid in vinegar production

[0042] Wheat bran 0.5kg, distilled water 10L, pH 4.8, oxygen flow 1vvm, rotation speed 150r / min, temperature 28°C, ferment, take the liquid fermentation liquid at the 76th hour of fermentation, use high performance liquid chromatography to measure ferulic acid esterase activity (such as image 3 Shown), the enzyme activity is 280U / L.

[0043] Chromatographic column: XBridgeTMC18 (4.6×250mm, 5μm)

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

[0045] Gradient elution method: starting time, the flow rate of solution B is 80%, the flow rate of solution D is 20%, and it is maintained for 2 minutes; 8 minutes, the flow rate of solution B is 72%, the flow rate of solution D is 28%; 13 minutes, the flow rate of solution B is 42%, the flow rate of solution D is 58 %; 19min, B solution flow ra...

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Abstract

The invention discloses Cladosporiumc cladosporioides for high yield of feruloyl esterase and an application of Cladosporiumc cladosporioides in edible vinegar brewing, and belongs to the field of fermentation engineering and biotechnology. The Cladosporiumc cladosporioides was preserved by the China Center for Type Culture Collection, the preservation number is CCTCC NO: M2015549, and the preservation date is September, 16th, 2015. According to the invention, wheat bran is used as a fermentation culture medium for fermentation production of high-activity feruloyl esterase, and it is easy to realize industrial production of feruloyl esterase. By applying feruloyl esterase to production of edible vinegar, functional health-care edible vinegar rich in ferulic acid is obtained. Thus, new technical support is provided for development of health-care vinegar.

Description

technical field [0001] The invention relates to 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. Background technique [0002] Ferulic acid esterase (FEA, EC3.1.1.73) is a subclass of carboxylate hydrolase, the main biological function is to hydrolyze the ester bond between plant cell wall polysaccharide and ferulic acid (FA), so that ferulic acid and other phenylacrylic acid released. At present, there are about 30 ferulic esterase-producing strains reported, mainly including Aspergillus niger, Aspergillus oryzae, Aspergillus terreus, Aspergillus flavus, Aspergillus flavusus, Aspergillus awamori, Aspergillus tabin, Aspergillus nidulans, Aspergillus Usami, Rhizopus, Penicillium, Sporosporium thermophilicum, Fusarium oxysporum, Fusarium laminarum, Neurospora crassa, Rhizomucor miehei, Basilicum stalk, Russula chlorophyll, Pleurotus eryngii, Aureob...

Claims

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Application Information

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Patent Type & Authority Applications(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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