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Bio-augmentation technology for fermentation process of solid-state brewing vinegar

A technology of bioaugmentation and fermentation process, applied in the field of bioengineering, can solve the problems that restrict the application of bioaugmentation technology and the problem of biological safety has not been well resolved

Active Publication Date: 2011-10-26
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reasons are as follows: ① The research on microbial strains in many traditional fermented foods has not been carried out in depth, and there is still little information about the composition and functions of the dominant bacterial groups that play a leading role in the colony structure. It is the premise of bioaugmentation research; ②Food safety issues have increasingly become the focus of attention in the field of food production. If exogenous strains or high-efficiency genetically engineered bacteria are used for bioaugmentation research, the potential biosafety problems have not been recognized so far. Well solved, this directly restricts the application of bioaugmentation technology in the traditional fermented food production industry

Method used

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  • Bio-augmentation technology for fermentation process of solid-state brewing vinegar
  • Bio-augmentation technology for fermentation process of solid-state brewing vinegar
  • Bio-augmentation technology for fermentation process of solid-state brewing vinegar

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Extraction of total DNA of vinegar fermented grain microbial community

[0030] 1. Sample pretreatment of vinegar fermented grains

[0031] Take 2g of vinegar unstrained spirits sample, suspend it with 20mL 0.1M PBS (pH 7.0), add glass beads, vortex fully shake for 5min; centrifuge at 200×g for 5min, collect the supernatant (including bacteria), discard the precipitate; wash 3 times; 9000×g high-speed centrifugation for 5 minutes. Discard the supernatant and collect the precipitate; suspend the collected bacteria in 5 mL of 0.1M PBS (pH 7.0), centrifuge at 9000×g; wash 3 times; suspend the washed bacteria in 1 mL of PBS (pH 7.0), blow with a gun After vortex, shake evenly, transfer to 1.5mLEP tube, and freeze at -20°C for later use.

[0032] 2. Extraction of genomic DNA from vinegar fermented grains

[0033] Weigh 2g of vinegar unstrained spirits, add liquid nitrogen into a mortar and grind thoroughly, transfer to a 50mL centrifuge tube. Add 6 mL of DN...

Embodiment 2

[0035] Example 2: PCR-DGGE technology analyzes the structure of the microbial community of vinegar fermented grains

[0036] 1. PCR amplification

[0037] bacterial 16S rDNA V 3 Primer P used in PCR amplification of region 2 -P3 Can amplify 16S rDNA V 3 A fragment with a region of about 196bp corresponds to the sites from 341 to 534 of E. coli16S rDNA.

[0038] 2. DGGE analysis

[0039] bacterial 16S rDNA V 3 The PCR amplified products were detected by 1% agarose gel electrophoresis and the DNA concentration was determined with DyNA QuantTM 200 concentration instrument (Hoefer Pharmacia Biotech). Electrophoresis was performed with the Bio-Rad DCode DGGE system. DGGE electrophoresis conditions are: 8% polyacrylamide gel, DGGE adopts 30%-50% denaturing gradient (100% denaturant concentration is 7M urea, 40% formamide), and the loading amount is 200ng. Electrophoresis buffer was 1×TAE, 60°C, 200V voltage electrophoresis for 4h, SYBR Green I staining for 45min, UVP2GDS8000 ...

Embodiment 3

[0044] Embodiment 3: Screening of vinegar-making functional microorganisms

[0045] Take 10g sample from Zhenjiang balsamic vinegar unstrained spirits and put it into a 500mL Erlenmeyer flask mixed with glass beads and 90mL sterile water, shake it on a shaker at 37°C at 115r / m for 30min, then take 1mL of the bacterial suspension and add sterile saline 10-fold serial dilution, with 10 -2 、10 -3 and 10 -4 Spread the gradient dilution sample on GYC plate and incubate at 37°C for 48-72 hours; judge whether it is acid-producing bacteria according to whether it has a transparent circle on the GYC plate, and then pick colonies with obvious transparent circle and abundant colonies, isolate and purify them, and insert them Slant culture medium, cultured at 37°C for 48h, and stored at 4°C.

[0046] Separation medium (GYC): glucose 1%, yeast powder 1%, peptone 0.3%, light CaCO 3 0.5%, 2% agar, sterilized at 0.1Mpa at 121°C for 20min, and then added 6% absolute ethanol (v / v) after coo...

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Abstract

The invention discloses a bio-augmentation technology for fermentation process of solid-state brewing vinegar, belonging to the technical field of biological Engineering. The operation procedures of the method comprise: (1) determining functional microorganisms for brewing vinegar: analyzing the structure of the microflora in vinegar fermented grains with molecular ecology technology such as PCR-DGGE, determining the functional microorganisms in the process of brewing vinegar by combining the key physicochemical indexes related to the quality and functionality of vinegar; (2) pure culture of the functional microorganisms for brewing vinegar: using microorganism pure culture technique to get the functional microorganisms for brewing vinegar from the vinegar fermented grains; and (3) bio-augmentation in the vinegar fermentation process: selecting the functional microorganisms or a combination thereof, and carrying out bio-augmentation in the solid-state fermentation stage of vinegar. The method has the advantages of shortening brewing cycle of vinegar, improving and controlling the flavor, quality and functionality of vinegar products, improving the utilization rate of raw material and the like.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a bioaugmentation technology used in the fermentation process of solid-state brewing vinegar. Background technique [0002] The vinegar industry is an important part of my country's biotechnology industry. In 2009, my country's vinegar output reached 4 million tons, ranking first in the world. The brewing methods of vinegar can be divided into two categories: solid state fermentation and liquid state fermentation. Most of the vinegar in our country adopts the traditional "solid-state open multi-strain mixed fermentation vinegar-making process", that is, the vinegar-making process in which the material is solid during acetic acid fermentation. The vinegar brewed by this method not only has a sour taste, but also has a unique flavor and health care function. During the fermentation process of vinegar, the formation of flavor substances such as acetic acid and f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12J1/00C12Q1/68C12Q1/04C12R1/02C12R1/125C12R1/225
Inventor 许正宏史劲松陆震鸣许伟
Owner JIANGNAN UNIV
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