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A culture method of Burkholderia polyphage and its application in catalytic synthesis of liquor flavor esters and degradation of liquor 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, poor quality of Luzhou-flavor liquor, etc., and achieve high application value

Active Publication Date: 2021-05-04
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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  • A culture method of Burkholderia polyphage and its application in catalytic synthesis of liquor flavor esters and degradation of liquor harmful esters
  • A culture method of Burkholderia polyphage and its application in catalytic synthesis of liquor flavor esters and degradation of liquor 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 invention belongs to the technical field of microorganisms, and in particular relates to a method for cultivating Burkholderia polyphagia CGMCC 1.3829 and its application in catalytically synthesizing liquor flavor esters and degrading liquor harmful esters. Through the cultivation method of the present invention, the bacterium has a high ability to efficiently catalyze the synthesis of liquor flavor esters ethyl caproate, ethyl caprylate and ethyl caprate in the aqueous phase system, and the experiments show that the yields can reach 61.90±8.04 mg / L, 746.27 mg / L and 746.27 mg / L respectively. ± 82.45mg / L and 783.91 ± 76.60mg / L, simultaneously according to the cultivation method of the present invention, it also has degrading liquor harmful ester dimethyl phthalate (DMP), diethyl phthalate (DEP), The ability of dibutyl phthalate (DBP) and bis(2-ethylhexyl) phthalate (DEHP), experiments show that the degradation rate can reach 19.07%±0.58%, 18.95%±0.41%, 36.33% respectively %±16.85%, and 53.76%±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 Patents(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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