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Culture method of burkholderia multivorans strain and applications of burkholderia multivorans in catalyzing synthesis of baijiu flavor esters and degradation of baijiu harmful esters

A technology that phagocytizes Burkholderia and catalyzes esters, applied in the field of microorganisms, can solve the problems of slow ester production, low synthesis efficiency, low yield of high-quality wine, etc., and achieve high application value

Active Publication Date: 2019-12-20
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the slow production of ester aroma during the brewing process, that is, the low synthesis efficiency of ethyl caproate, ethyl caprylate and other esters, the brewing cycle of aroma generation is long and the yield of high-quality wine is low, while ethyl caproate and other important Low ester content has always been one of the key reasons for the poor quality of Luzhou-flavor liquor and the low yield of high-quality liquor

Method used

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  • Culture method of burkholderia multivorans strain and applications of burkholderia multivorans in catalyzing synthesis of baijiu flavor esters and degradation of baijiu harmful esters
  • Culture method of burkholderia multivorans strain and applications of burkholderia multivorans in catalyzing synthesis of baijiu flavor esters and degradation of baijiu harmful esters

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Nitrogen source and carbon source optimization of Burkholderia polyphage CGMCC 1.3829 fermentation culture

[0027] This example is carried out in order to optimize the cultivation method of Burkholderia polyphage CGMCC 1.3829 to obtain better liquor flavor esters.

[0028] 1. Strain activation: Under aseptic conditions, inoculate Burkholderia polyphage CGMCC 1.3829 into a 30mL test tube containing 5mL fermentation medium, and culture on a shaker at 30±2°C, 150±50r / min for 1 -2 days.

[0029] 2. Nitrogen source optimization fermentation culture: Under aseptic conditions, inoculate activated Burkholderia polyphagocytida CGMCC1.3829 into a 300mL Erlenmeyer flask containing 100mL fermentation medium, shaker at 30±2°C, 150± 50r / min, cultivate for 2–5 days. The fermented liquid after cultivating is prepared crude enzyme liquid according to embodiment 2 (1st point), and gained crude enzyme liquid carries out the mensuration of ester synthesis according to embodime...

Embodiment 2

[0033] Example 2 Preparation of Burkholderia polyphage CGMCC 1.3829 Crude Enzyme Preparation and Catalytic Ester Synthesis under Conditions of Simulated Liquor Fermentation Water Phase System

[0034] Based on the culture solution obtained in Example 1, the following experiments were carried out.

[0035] 1. Preparation of crude enzyme preparation:

[0036] Take 10-30 mL of the fermentation medium in a 50 mL centrifuge tube, break up the bacterial cells with an ultrasonic cell disruptor, centrifuge at 6000×g for 10 min, and take the supernatant as a crude enzyme preparation.

[0037] 2. Catalyzing ester synthesis under the condition of simulated liquor fermentation water phase system

[0038] The 10mL reaction system is as follows: Burkholderia polyphagia CGMCC 1.3829 crude enzyme solution, 1mL; citric acid buffer (pH 4.0), 9mL (add ethanol to 1M); caproic acid, caprylic acid and capric acid, the final concentration 10mM. 30±1°C, 150±10r / min water bath shaker reaction for 1...

Embodiment 3

[0044] Example 3 Degradation of Burkholderia polyphage CGMCC 1.3829 to DMP, DEP, DBP and DEHP

[0045] In the experiment, it was found that Burkholderia polyphage CGMCC 1.3829 has the property of degrading some harmful esters, so this property was further verified based on the following culture method. The composition of the medium includes: yeast powder 5.0 g / L, (NH 4 ) 2 SO 4 2.0g / L, MgSO 4 ·7H 2 O 0.2g / L, CaCl 2 2H 2 O 0.01g / L, FeSO 4 ·7H 2 O 0.001g / L, Na 2 HPO 4 12H 2 O 1.5g / L, KH 2 PO 4 1.5g / L, sterilized at 115°C for 20min, and added DMP, DEP, DBP and DEHP with a final concentration of 200mg / L. Put 10mL of medium into a 100mL Erlenmeyer flask, and inoculate 1–3%. The culture conditions are: 30±2°C, 150±50r / min, culture for 3-7 days.

[0046] After culturing, transfer 10 mL of fermentation broth into a 50 mL centrifuge tube, add 2 mL of n-hexane, shake and mix vigorously for 30 seconds, centrifuge, take the supernatant, filter and centrifuge for quantitati...

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Abstract

The present invention belongs to the technical field of microorganisms and particularly relates to a culture method of burkholderia multivorans CGMCCCY 1.3829 and applications of burkholderia multivorans in catalyzing synthesis of baijiu flavor esters and degradation of baijiu harmful esters. The bacterium cultured by the method has high ability to catalyze the synthesis of the baijiu flavor esters of ethyl hexanoate, ethyl octoate, and ethyl decanoate in water phase systems. Experiments show that yield of the ethyl hexanoate can reach 61.90 plus or minus 8.04 mg / L, yield of the ethyl octoatecan reach 746.27 plus or minus 82.45 mg / L and yield of the ethyl decanoate can reach 783.91 plus or minus 76.60 mg / L, respectively. At the same time, according to the culture method, the burkholderiamultivorans also has ability to degrade the baijiu harmful esters of dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP) and di-(2-ethylhexyl) phthalate (DEHP). Experiments showthat a degradation rate of the dimethyl phthalate can reach 19.07% plus or minus 0.58%, a degradation rate of the diethyl phthalate can reach 18.95% plus or minus 0.41%, a degradation rate of the dibutyl phthalate can reach 36.33% plus or minus 16.85%, and a degradation rate of the di-(2-ethylhexyl) phthalate can reach 53.76% plus or minus 17.60%.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a method for cultivating Burkholderia polyphagia and its application in catalyzing the synthesis of flavor esters of liquor and / or the degradation of harmful esters of liquor. [0002] technical background [0003] The main components of liquor are water and ethanol, but what determines the quality and style of liquor is the trace aroma and taste components with a content of 1%-2%, including esters, alcohols, acids, aldehydes and ketones, heterocyclic compounds, carbonyls, etc. Among them, esters are the flavor compounds with the largest variety and highest content found in liquor so far. It is reported that there are more than 400 kinds of them, and their content accounts for 75%-95% of the flavor components of liquor. In addition, esters with fruity, floral and sweet aromas not only endow the liquor with a unique aroma, but also influence and even determine th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/62C12G3/02C12R1/01
CPCC12N1/20C12G3/02Y02E50/10
Inventor 李秀婷徐友强孙宝国王晓程
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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