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Leuconostoc mesenteroides and application thereof

A technology of Leuconostoc membranaceus and bacterial agent, which is applied in the field of microorganisms and can solve the problems of poor product safety, unstable quality, and limitation of large-scale production and application.

Pending Publication Date: 2021-07-23
四川老坛子食品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the increase in the content of 2-methylhexanoic acid achieved through the addition of exogenous sources leads to poor safety and unstable quality of the product due to the risk of impurities in the exogenous additives
Therefore, the large-scale production and application of exogenous addition of 2-methylhexanoic acid in improving the flavor of fermented foods is limited.

Method used

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  • Leuconostoc mesenteroides and application thereof
  • Leuconostoc mesenteroides and application thereof
  • Leuconostoc mesenteroides and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Isolation method of Leuconostoc mesenteroides strain

[0061] 1. Obtain a suitable dilution gradient and culture

[0062] Sampling from traditional kimchi (commercially available) in Meishan City, Sichuan Province, weighed 5g, added to 45mL sterile water, and took 4 dilutions of 10 after gradient dilution. -1 , 10 -2 , 10 -3 , 10 -4 100 μL of each of the bacterial suspensions were spread on the MRS agar medium and incubated at 37°C for 48 hours.

[0063] 2. Separation and purification

[0064] Use the plate streaking method to select typical single colonies, and repeat the culture selection operation to obtain strains with excellent traits.

[0065] 3. Primary screening of strains

[0066] Gram stain and catalase test

[0067] Pick a single colony and do Gram staining and catalase experiments. Gram-positive and hydrogen peroxide-negative bacteria are purified on a plate, centrifuged at 10,000rpm for 5min, and stored in a 30% (v / v) glycerol tube. - Store at 80°C....

Embodiment 2

[0094] Identification of Leuconostoc mesenteroides B-MS-PC-13

[0095] 1. PCR amplification of 16SrDNA

[0096] The Leuconostocmesenteroides (Leuconostocmesenteroides) B-MS-PC-13 single bacterial colony obtained in the preliminary screening process of the bacterial strain in step 3 of Example 1 was planted on the MRS Qiongzhou plate, cultivated at 37°C for 18 to 24 hours, and the grown single Colonies were used as templates for PCR.

[0097] 1) 25 μL of PCR system, including 12.5 μL of Mix, 0.5 μL of 27F, 0.5 μL of 1492R, and 11.5 μL of ddH2O.

[0098] The primers used were 27F: AGAgTT TGA TCC TGG CCT CA and 1492R: gGT TAC CTTgTTACGACTT. The length of the amplified fragment is 1500bp.

[0099] 2) PCR conditions:

[0100] DNA double strands were pre-denatured at 94°C for 10 minutes; denatured at 94°C for 30s; annealed at 50°C for 30s; extended at 72°C for 80s, and cycled 29 times, and finally kept at 72°C for 7 minutes.

[0101] 2. Agar gel electrophoresis

[0102] Weigh ...

Embodiment 3

[0111] Growth and Fermentation Characteristics of Leuconostoc mesenteroides

[0112] (1) Growth curve of Leuconostoc mesenteroides strain

[0113] The Leuconostoc mesenteroides B-MS-PC-13 of Example 1 stored at -80°C was inoculated in MRS broth medium and cultured at 37°C for 24 hours to obtain an activated strain.

[0114] Inoculate the activated bacteria into the MRS broth medium according to the inoculum amount of 2-4% (v / v), culture at a constant temperature of 37°C for 26 hours, measure the OD value of the culture solution at 600nm every 2 hours, and use the OD value as a function of time. The figure obtains the growth curve of the bacterial strain, and the results are shown in Figure 4 (Growth graph of Leuconostoc mesenteroides). The result shows: show: enterofilm Leuconostoc mesenteroides (Leuconostoc mesenteroides) B-MS-PC-13 grows rapidly in the MRS medium, after 18h, reaches the maximum amount of bacteria 6.8*10 9 CFU / mL. , the lag phase of the strain is relativ...

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Abstract

The invention discloses Leuconostoc mesenteroides and application thereof. The Leuconostoc mesenteroides is preserved in Guangdong Microbiological Culture Collection Center, and the preservation number is GDMCCNO. 61344; and the Leuconostoc mesenteroides is a safe strain capable of being used for food, the yield of 2-methylhexanoic acid is high, the flavor of fermented food can be improved, and the strain is suitable for large-scale production and application.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Leuconostoc enterococcus and application thereof. Background technique [0002] 2-methylhexanoic acid (2-methylhexanoic acid, C 7 h 14 o 2 ), is one of the important flavor compounds in fermented food, usually endows cheese, red wine, pickles and other fermented foods with creamy and lipid aroma. When its concentration reaches 0.9μg / kg, it can contribute the unique aroma of fermented vegetables in terms of flavor sensory quality, making the aroma of fermented vegetables more mellow and harmonious. Usually, the yield of 2-methylhexanoic acid in the vegetable fermentation process is low. In production, in order to achieve the purpose of flavor enhancement, the flavor of fermented vegetable products can be improved by adding exogenous sources. [0003] However, the increase in the content of 2-methylhexanoic acid achieved through the addition of exogenous sources has po...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04C12P7/40A23L19/20A23L2/38A23L2/84A23L11/50A23L29/00A23C9/123C12R1/01
CPCC12N1/20C12N1/04C12P7/40A23L19/20A23L2/382A23L2/84A23L29/065A23C9/1234A23V2400/321
Inventor 何艳平赵楠
Owner 四川老坛子食品有限公司
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