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Method for reducing ethyl carbamate in fermented food by synthesizing leavening agent

A starter and Lactobacillus fermentum technology are applied in the field of synthesizing starter to reduce urethane in fermented food, and can solve problems such as difficulty in regulating urea metabolism

Active Publication Date: 2022-03-01
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is difficult to regulate urea metabolism only by engineering of S. cerevisiae strains

Method used

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  • Method for reducing ethyl carbamate in fermented food by synthesizing leavening agent
  • Method for reducing ethyl carbamate in fermented food by synthesizing leavening agent
  • Method for reducing ethyl carbamate in fermented food by synthesizing leavening agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1: Screening and isolation of Saccharomyces cerevisiae G2

[0081] Pre-screening: Weigh 1g of yellow rice wine fermented mash sample into a sterile EP tube, mix thoroughly with 1mL of sterile saline, dilute to an appropriate multiple, take 100μL, and spread on YNB solid medium containing urea as the only nitrogen source , placed in a constant temperature incubator at 30°C and cultivated until a single colony grows on the plate, and picked by QPix420 biological screening system or manually.

[0082] Primary screening: Use the microbial screening system QPix420 or manual selection to transfer a single colony on the pre-screening plate to a 48-well plate containing the primary screening medium, and then culture it in an orifice shaker at 30°C and 220r / min After 48h, centrifuge the 48-well plate at 3500r / min for 5min, take 80μL of the supernatant in a 96PCR plate, add 20μL of diacetylmonoxime thiamine and 100μL of ferric ammonium phosphate solution, react at 100°C f...

Embodiment 2

[0084] Embodiment 2: Screening and isolation of Rhizopus oryzae (Rhizopus oryzae) M5

[0085] Weigh 1g of rice wine fermented mash and put it in a sterile EP tube, take 1mL of sterile saline and mix well, dilute to an appropriate multiple, take 100μL and spread it on the PDA culture, and cultivate it in a constant temperature incubator at 28°C for 2-3 days. By measuring the glucoamylase activity in PDA, the strains with high glucoamylase activity were screened out, and the strains were preserved at 4°C.

[0086] Finally, a strain of mold M5 was obtained.

Embodiment 3

[0087] Embodiment 3: the screening of producing urease and EC hydrolase bacterial strain lactic acid bacteria d6

[0088] Preliminary screening: Weigh 1g of rice wine fermented mash into a sterile EP tube, take 1mL of sterile saline and mix well, dilute to an appropriate multiple, take 100μL and apply it to the separation culture, and cultivate it in a constant temperature incubator at 37°C for several days until a single colony grows. Use the QPix420 screening system to screen bacteria, pick a single colony into a 48-well plate with screening medium, and culture it anaerobically at 37°C for several hours. Since urease or EC hydrolase can react with the corresponding substrate to produce ammonia, the bromocresol purple changes from purple to yellow. According to the color reaction, select the target strain for primary screening.

[0089] Re-screening: select the target strain of primary screening and inoculate it in MRS medium for 72 hours, take the bacterial liquid in a refr...

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Abstract

The invention discloses a method for reducing ethyl carbamate in fermented food through a synthetic leavening agent, and belongs to the field of wine brewing and food safety. The invention provides a strain of lactobacillus fermentum d6 which is preserved in the China Center for Type Culture Collection on November 2, 2021, and the preservation number of the lactobacillus fermentum d6 is CCTCC NO: M 20211358. A synthetic fermentation agent containing lactobacillus fermentum, saccharomyces cerevisiae, rhizopus oryzae and Vickerhamomyces anomalus is prepared to simulate yellow rice wine fermentation, and the result shows that the content of urea and the content of ethyl carbamate in the yellow rice wine after fermentation are detected to be obviously reduced compared with a control group, and the maximum reduction amount is 97.4% and 90.9% respectively.

Description

technical field [0001] The invention relates to a method for synthesizing a leavening agent to reduce ethyl carbamate in fermented food, belonging to the fields of wine brewing and food safety. Background technique [0002] Ethyl carbamate (EC) is classified as a class 2A carcinogen and is present in various fermented foods such as vinegar, soy sauce, pickles, cheese, chocolate, and alcoholic beverages. Since EC in alcoholic beverages is very harmful to the health of consumers, it is necessary to reduce the concentration of EC in these fermented foods and alcoholic beverages. [0003] Rice wine is usually brewed from rice, wheat koji and Saccharomyces cerevisiae. During the brewing process of rice wine, fungi and yeast are important active microorganisms in the rice wine starter, which are responsible for starch degradation and alcohol fermentation, respectively. In alcoholic beverages, the reaction of urea with ethanol is the most common route of EC generation. Urea was ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/18C12N1/16C12N1/14C12G3/024C12G3/02C12G3/022C12G1/022C12H6/02C12R1/25C12R1/865C12R1/845C12R1/645
CPCC12N1/20C12N1/18C12N1/16C12N1/14C12N9/80C12Y305/01075C12G3/024C12G3/02C12G3/022C12G1/0203C12H6/02C12G2200/05
Inventor 余世琴周景文田淑芳夏小乐曾伟主陈坚堵国成
Owner JIANGNAN UNIV
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