A strain of Lactococcus lactis with high yield of 6-phospho-β-galactosidase and its application

A Lactococcus lactis, milk technology, applied in the field of microorganisms, can solve the problems of low yield of 6-phosphate-β-galactosidase, affecting the quality of fermented dairy products, etc., and achieves reduction of lactose content, low lactose content, and growth rate quick effect

Active Publication Date: 2020-08-04
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are mainly two lactose metabolic pathways in lactic acid bacteria. The key enzymes of these two metabolic pathways are β-galactosidase and 6-phospho-β-galactosidase. Among them, β-galactosidase is common in lactic acid bacteria. However, 6-phospho-β-galactosidase is only found in a few kinds of lactic acid bacteria (mainly including Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactococcus lactis subsp. cocci), and most of the 6-phospho-β-galactosidase-producing lactic acid bacteria found so far have low 6-phospho-β-galactosidase production, which is not enough to affect fermented dairy products The point of quality

Method used

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  • A strain of Lactococcus lactis with high yield of 6-phospho-β-galactosidase and its application
  • A strain of Lactococcus lactis with high yield of 6-phospho-β-galactosidase and its application
  • A strain of Lactococcus lactis with high yield of 6-phospho-β-galactosidase and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1: Screening and identification of bacterial strains

[0059] 1. Screening

[0060] (1) Prepare appropriate sample dilution gradient and culture

[0061] Take out the Qinghai Xining traditional fermented dairy product triglycerin sample stored in the -80℃ refrigerator, put it on ice to thaw, shake and mix, take 0.5mL sample and add it to 4.5mL sterile normal saline, complete a 10-fold dilution, shake After mixing evenly, take 0.5mL of the diluted solution and add it to 4.5mL sterile saline to complete the second 10-fold dilution, and so on until the dilution reaches 10. -6 , draw 100 μL from each gradient dilution, evenly spread it on the MRS solid medium plate, invert it, place it under anaerobic culture at 37°C for 36-48 hours, and observe it in time.

[0062] (2) Line separation and purification

[0063] After taking out the plate with colonies, select a gradient plate with obvious single colonies, pick colonies with different colony shapes, and perform...

Embodiment 2

[0085] Embodiment 2: the cultivation of bacterial strain

[0086] Lactococcus lactis (Lactococcus lactis) CCFM1033 was inoculated on the MRS solid medium, cultured upside down at 30°C for 48 hours, and the colony morphology was observed; After mordanting, washing, decoloring, counterstaining, washing, and drying, microscopic examination was performed, and the morphology and Gram staining results were recorded.

[0087] The Lactococcus lactis CCFM1033 observed under the oil microscope is spherical, and the Gram staining result is purple, which means it is a Gram-positive bacterium (see figure 1 ); And, the bacterial colony of this bacterial strain on the solid MRS medium is white, glossy, with smooth edges, opaque, slightly raised, and medium in size (see figure 2 ).

Embodiment 3

[0088] Example 3: Growth characteristics of bacterial strains in dairy systems

[0089] 1. The growth curve of Lactococcus lactis grown in skim milk for 12 hours

[0090] Lactococcus lactis (Lactococcus lactis) CCFM1033 and Lactococcus lactis (Lactococcus lactis) D-XJ 4-12 stored at -80°C were respectively inoculated in MRS liquid medium, cultured at 30°C for 24 hours, subcultured 2 to 3 times , until the bacterial concentration reaches 10 8 ~10 9 CFU / mL; the bacteria solution activated in MRS was taken out and inoculated in skim milk at a volume ratio of 2% to 4%, so that the amount of bacteria in the system reached 10 7 CFU / g; put the inoculated sample into an incubator at 37°C for fermentation, take samples every 2 hours, and detect the changes in the amount of bacteria during the fermentation process. The results are as follows: image 3 shown.

[0091] Depend on image 3 It can be seen that the bacterial count of Lactococcus lactis (Lactococcus lactis) CCFM1033 reached...

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Abstract

The invention discloses a strain of Lactococcus lactis capable of high-yielding 6-phosphate-β-galactosidase and an application thereof, belonging to the technical field of microorganisms. This Lactococcus lactis (Lactococcus lactis) CCFM1033 has the advantages of fast growth rate, high yield of 6-phosphate-β-galactosidase and good aroma-producing properties, can effectively reduce the lactose content in fermented dairy products, and is beneficial to fermented dairy products The improvement of quality has a very important application in the preparation of fermented milk products.

Description

technical field [0001] The invention relates to a strain of Lactococcus lactis high-yielding 6-phospho-β-galactosidase and application thereof, belonging to the technical field of microbes. Background technique [0002] Fermented dairy products are acidic dairy products made from milk as the main raw material, fermented by lactic acid bacteria or co-fermented by lactic acid bacteria and yeast. As a kind of milk product, it is deeply loved by people for its unique taste and flavor, and has been sold in the market for many years. . Studies have shown that the main factors affecting consumers' purchase of fermented dairy products are flavor, price, availability and brand, among which flavor accounts for the most weight. Therefore, improving the flavor of fermented dairy products is undoubtedly the most important means for companies to cater to the market. [0003] Lactose is the most important carbohydrate in milk, accounting for more than 99% of the total carbohydrates in mil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12G3/026A23C9/123C12R1/01
CPCA23C9/123C12G3/02C12N1/20C12R2001/46C12N1/205A23V2400/157
Inventor 陈卫刘小鸣杨宇王鸿超赵建新张灏
Owner JIANGNAN UNIV
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