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New lactic acid bacteria

A strain and amino acid technology, applied in bacteria, dairy products, microbial-based methods, etc., can solve problems such as expensive and complicated manufacturing processes

Pending Publication Date: 2022-02-18
DUPONT NUTRITION BIOSCIENCES APS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these solutions are unsatisfactory for dairy manufacturers as they require the addition of exogenous enzymes (such as lactase) to the milk prior to fermentation, making the manufacturing process more complex and expensive, or adding Added carbohydrates (such as sucrose), which is inconsistent with the growing demand for healthier products without additives

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0319] Example 1: Isolation of S. thermophilus exhibiting a full-stop phenotype.

[0320] Dilutions of cultures of the DGCC7984 strain were plated on the surface of M17 supplemented with 5 g / L sucrose agar plates. After incubation at 37°C for 48 hours, two isolated colonies of the DGCC7984 strain were picked and propagated in M17 liquid medium supplemented with 20 g / L sucrose at 37°C for 24 hours. These two subclones of the DGCC7984 strain were named DGCC12455 and DGCC12456. The acidifying properties of strains DGCC12455 and DGCC12456 were investigated as follows: both strains were inoculated into M17 liquid medium supplemented with 30 g / L lactose and then incubated overnight at 37°C. The culture was washed (v / v) in tryptone saline solution (tryptone 1 g / L, NaCl 8.5 g / L) as follows: the culture was centrifuged at 4000 rpm for 5 min; the pellet was resuspended in 10 mL of tryptone saline in solution. The washed culture was inoculated at 1% (v / v) into 100 mL of UHT semi-skimm...

Embodiment 2

[0322] Example 2: DGCC12456 lacZ Identification of Genetic Differences in Genes

[0323] The genomes of strains DGCC7984 and DGCC12456 were sequenced and compared. Among them, in lacZ Differences between the two strains were identified in genes. lacZ Genes have been described (van den Bogaard et al., 2000; Vaughan et al., 2001) to encode β-galactosidase, the enzyme responsible for the hydrolysis of lactose into glucose and galactose. In the DGCC12456 genome, lacZ The C base at position 1060 of the gene was replaced with a T base, resulting in a non-conservative amino acid change, and the arginine at position 354 of β-galactosidase was replaced with a cysteine ​​(R354C substitution). Therefore, DGCC7984 has a β-galactosidase-encoding lacZ Alleles, the sequence of the β-galactosidase is as defined in SEQ ID NO: 2, and the DGCC12456 strain has the sequence encoding the β-galactosidase lacZ Alleles, the sequence of the β-galactosidase is as defined in SEQ ID NO:4. In contra...

Embodiment 3

[0328] Example 3: Comparison of the acidification curves of S. thermophilus strains DGCC715 and DGCC11231 and their derivatives encoding a β-galactosidase having the sequence SEQ ID NO:4 instead of SEQ ID NO:2 (R354C substitution).

[0329] Builds are named 715 respectively R354C and 11231 R354C Derivatives of strains DGCC715 and DGCC11231. Inserted into the DGCC12456's lacZ Gene (encoding beta-galactosidase with cysteine ​​(C) at position 354) to replace that of strains DGCC715 and DGCC11231 lacZ Gene. Indeed, from DGCC12456 DNA PCR amplified lacZ Gene. Competent cells of DGCC715 or DGCC11231 were prepared and transformed with amplified DNA. Transformants were verified by sequencing.

[0330] Evaluation of Streptococcus thermophilus strains DGCC715, DGCC11231, 715 R354C and 11231 R354C Capacity to ferment milk as described in the Materials and methods section [Assay C]. The pH was recorded over time using a CINAC device, and the results are presented at figure 2 A...

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Abstract

The invention relates to a polynucleotide comprising a lacZ gene (lacZ FS ) encoding a [beta]-galactosidase characterized by a particular profile regarding its efficiency of hydrolysis of lactose. The invention is also directed to a Streptococcus thermophilus strain comprising a lacZ FS allele and bacterial composition thereof, and their use to obtain fermented milk not undergoing post-acidification.

Description

[0001] field of invention [0002] The present invention relates to a protein comprising an encoding β-galactosidase lacZ Gene( lacZ FS ), said β-galactosidase is characterized by a specific profile with respect to its lactose hydrolysis efficiency. The invention also relates to the inclusion of lacZ FS Allelic Streptococcus thermophilus strains and bacterial compositions thereof, and their use for obtaining fermented milks that do not undergo post-acidification. [0003] Background of the invention [0004] The food industry uses bacteria to improve the taste and texture of food or feed products. In the case of the dairy industry, lactic acid bacteria are often used, for example, to achieve acidification of milk (by fermentation of lactose) and to texturize products into which they are incorporated. For example, Streptococcus thermophilus ( S. thermophius ) species of lactic acid bacteria are widely used alone or in combination with other bacteria for the manufacture o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/56C12N9/38C12N1/21C12N15/74C12P7/56A23C9/12C12R1/46
Inventor A·叶德烈约夫斯基C·弗雷莫S·范迪伦T·德斯福格雷M·C·若多D·卢根
Owner DUPONT NUTRITION BIOSCIENCES APS
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