Malolactic fermentation (MLF) lactic acid bacteria strain with high yield of esterase and application thereof

A technology of lactic acid fermentation and malic acid is applied in the field of bioengineering to achieve the effects of excellent fermentation performance, increased total amount, and improved fruit flavor

Active Publication Date: 2020-09-15
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no promotion and application of excellent local O.oeni in China. Enterprises either do not carry out MLF in the production process, or they can only choose commercial starters imported from abroad for fermentation, resulting in serious problems of standardization and homogeneity of the wine produced.

Method used

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  • Malolactic fermentation (MLF) lactic acid bacteria strain with high yield of esterase and application thereof
  • Malolactic fermentation (MLF) lactic acid bacteria strain with high yield of esterase and application thereof
  • Malolactic fermentation (MLF) lactic acid bacteria strain with high yield of esterase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A high-yield esterase malate-lactic acid fermenting lactic acid bacteria strain GS-2 in this example was obtained by isolation and screening from naturally initiated malic acid-lactic acid fermentation wine samples in the Hexi Corridor wine producing area of ​​Gansu Province. The specific isolation and screening method is to collect The production company naturally starts the malo-lactic acid fermentation of the snake dragon pearl wine sample, first diluted with 0.9% NaCl sterilized water to 10 -4 -10 -8 , after shaking well, draw 0.1mL of diluent, spread on ATB separation medium (containing 50mg / L cycloheximide) plates respectively, and culture anaerobically at 28°C for 5-7d. After the colony is formed, pick a white single colony with a diameter of less than 1 mm, a smooth surface, and a moist surface, and repeatedly streak and separate it on the ATB isolation medium until it is purified. The primary screening strains were inoculated into 48-well plates containing thr...

Embodiment 2

[0038] In this example, Oenococcus oeni (O.oeni) GS-2 was used as the test strain, and the commercial strain VP41 was used as the control, and its growth curve in ATB medium was determined. The specific application process is as follows:

[0039] ATB culture medium formula is the same as embodiment 1.

[0040] Strain activation:

[0041] Take the cryococcus oeni strain GS-2, put it at room temperature for 2 hours, pick 2 loops from the slant medium and inoculate it into the prepared ATB liquid medium, culture it in an incubator at 28°C, and set aside.

[0042] VP41 bacterial powder was activated with sterile distilled water. According to the added amount of 0.02g / L, the Oenococcus oeni VP41 strain was added to sterile dd H2O for 15 minutes at 20°C, and then an equal volume of ATB medium was added for 20 minutes at 25°C.

[0043] Biomass determination:

[0044] O.oeni activated to logarithmic growth phase was inoculated at 107 CFU / mL into an Erlenmeyer flask filled with 150 ...

Embodiment 3

[0048] In this example, Oenococcus oeni (O.oeni) GS-2 was used as the test strain, and the commercial strain VP41 was used as the control to measure the ability of the strain to degrade L-malic acid. The specific application process is as follows:

[0049] Bacterial strain activation: with embodiment 2.

[0050] Wine simulation juice: glucose 1g / L, fructose 1g / L, trehalose 1g / L, tartaric acid 1g / L, L-malic acid 3g / L, citric acid 1g / L, sodium acetate 0.14g / L, yeast extract powder 4.0 g / L, hydrolyzed casein 2.5g / L, KH 2 PO 4 0.3g / L, KCl0.22g / L, L-cysteine ​​hydrochloride 0.5g / L, MgSO 4 ·7H 2 O 0.065g / L, MnSO 4 4H 2 O 0.015g / L, CaCl 2 0.065g / L.

[0051] L-malic acid determination:

[0052] The Gernischt grapes collected from the grape planting base of Gansu Wuwei Mogao Wine Industry Co., Ltd. in 2019 were fermented according to the dry red wine production process, and poured into 2.5L brown fermentation bottles. The activated GS-2 strain was treated with 10 7 The CFU / m...

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Abstract

The invention discloses a malolactic fermentation (MLF) lactic acid bacteria strain with high yield of esterase and application thereof. The strain is classified and named as Oenococcus oeni (O.oeni)GS-2 collected in the China General Microbiological Culture Collection center (CGMCC) on October 14, 2019 with the collection number of CGMCC 18683. A grape wine sample is inoculated with the O.oeni GS-2 after alcohol fermentation is finished; and MLF can be completed within 12-14 d under the condition that the fermentation temperature is 18-20 DEG C, the fermentation perofrmnace is excellent, thetotal amount of ester aroma compounds in the fermented wine sample can be remarkably increased, the fruit aroma is improved, dry red wine with a typical style is brewed, and good economic benefits and application prespects are achieved.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a malic acid-lactic acid fermenting lactic acid bacteria strain with high esterase production and application thereof. Background technique [0002] Aroma quality is one of the most critical factors for consumers to choose and evaluate wine. Volatile compounds such as higher alcohols, esters, organic acids, aldehydes and ketones produced by the metabolism of yeast and lactic acid bacteria during the fermentation process are the main components of wine fermentation aroma. Esters account for about 30% of the volatile substances in fermentation, and most of them have elegant fruity and floral aromas, and their content is around the threshold. When there are slight changes in their composition, content and proportion, they may directly determine the quality of the wine. The aroma type and elegance of the body make the wine show different personality characteristics. Accordin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12G1/022C12R1/01
CPCC12G1/0203C12G2200/05C12R2001/01C12N1/205
Inventor 祝霞杨学山赵丹丹王璐璐王诗赵芳琴韩舜愈
Owner GANSU AGRI UNIV
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