Lactobacillus helveticus bacterium having high capability of producing tripentide, fermented milk product, and preparing process thereof

A technology for dairy products and Lactobacillus, applied in the field of new lactic acid bacteria Lactobacillus helveticus

Inactive Publication Date: 2001-01-10
CALPIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of the currently known lactic acid bacteria can produce high-efficiency lactotripeptides

Method used

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  • Lactobacillus helveticus bacterium having high capability of producing tripentide, fermented milk product, and preparing process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] (Selection of strains capable of producing fermented milk with high ACE inhibitory activity)

[0103] 36 strains of Lactobacillus helveticus isolated from different milk products were screened. The ACE inhibitory activity in the fermented milk of each strain was determined according to the following method. Each Lactobacillus helveticus was cultured at 37° C. for 24 hours in a 9% (weight percent) non-fat milk solids medium. The cultivated medium was added to the same type of fresh medium, and the new medium contained 3% (weight percentage) of the cultivated medium. It was further fermented at 37°C for 24 hours. After the fermentation, the acidity of lactic acid (weight percent), peptide content in whey (μg / mL), cell number and ACE inhibitory activity (U / mL) were measured. The results are shown in Table 1. Among the 36 strains, 7 strains had extremely weak fermentation ability. The acidity of lactic acid in the fermented milk produced by 15 strains is not lower than 1...

Embodiment 2

[0129] Select 11 strains from the 36 strains of Lactobacillus helveticus in Example 1, extract chromosomal DNA with reference to methods such as Leenhouts (Leenhouts K (1990) Applied Environmental Microbiology 56: 2726), digest with several restriction enzymes, at 0.8 Electrophoresis band patterns were analyzed on % agarose gel electrophoresis.

[0130] As a result, characteristic DNA fragments could be observed in the chromosomal DNA fragments of CM4 strain digested with EcoRI and PsII (see arrow 1 in Figure 1). It was not observed in the chromosomal fragments of the remaining strains, but a fragment shorter than the characteristic fragment of CM4 could be observed in most strains (see arrow 2 in Figure 1). The digested product of lambda bacteriophage DNA HindⅢ was used as the molecular size marker (23.1kb, 9.4kb, 6.6kb, 4.4kb, 2.3kb, 2.0kb, and the mobility was accelerated in turn) to compare the electrophoresis results, and the characteristic fragments The molecular weight...

Embodiment 3

[0132] Produce fermented milk with Lactobacillus helveticus CM4 selected in Example 1. The CM4 strain was cultured in 100 g of 9% (weight percent) skim milk at 37° C. for 12 hours, then mixed with 3 kg of fresh medium and continued to culture at 37° C. for 12 hours. After fermentation, all fermented milk (CM4 cell number 6.3×10 8 / mL) as 100kg of 9% (weight percent) skim milk starter and cultivated at 32°C for 20 hours. After the fermentation, the fermented milk contained 74.8 μg / mL lactotripeptide, and the lactic acid content was 1.9% (percentage by weight).

[0133] Take 43kg of fermented milk which has been prepared, homogenate with 4kg of granular sucrose, 3kg of water and 0.15kg of high methoxy pectin to prepare 50kg of yogurt drink. The yogurt beverage has a good taste, pH 3.6, and the number of live CM4 cells is 4.6×10 8 / mL.

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Abstract

There is provided a lactic acid bacteria capable of producing a large amount of lactotripeptide; also a fermented milk product that contains a large amount of active ingredient having hypotensive activity and anti-stress effect and can be taken pleasantly, is provided. More specifically, the present invention relates to Lactobacillus helveticus, that is, Lactobacillus helveticus CM4 strain (deposited at National Institute of Bioscience and Human-Technology Agency of Industrial Science and Technology, deposition number FERM BP-6060), characterized in having specific bacteriological properties, the bacteria, when cultured in a medium of animal milk containing 9 wt % solid of non-fat milk, producing tripeptides Val-Pro-Pro and Ile-Pro-Pro in an amount of 60 mu g in terms of Val-Pro-Pro per ml medium, and the bacteria exhibiting extracellular proteinase activity of not lower than 400U / OD590. The present invention also relates to a fermented milk product obtained by fermenting an animal milk with these lactic acid bacteria.

Description

technical field [0001] The invention relates to a novel lactic acid bacterium Lactobacillus helveticus, which can efficiently produce a unique tripeptide when cultured in animal milk medium, and has high extracellular protease activity; fermented milk products containing the lactic acid bacterium; the Method of preparation of dairy products. technical background [0002] Lactobacillus helveticus has long been used as a typical lactic acid bacteria starter for dairy products to produce fermented milk. Lactobacillus helveticus has high proteolytic activity, especially its highly active extracellular protease plays an important role in animal milk fermentation, that is, extracellular protease digests animal milk protein to produce various peptide fragments. The generated peptide fragments are further generated into low molecular weight peptides by the action of peptidases. It is well known that due to the action of protease, a part of peptides produced in the culture medium e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23C9/123A23C9/137C12N1/20C12P21/02C12R1/225
CPCC12N1/20A23Y2220/39C12R1/225A23C9/137A23C9/1234C12P21/02C12R2001/225C12N1/205A23V2400/147
Inventor 山本直之河上夏江石田优屋田裕一
Owner CALPIS
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