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Bacterium lactobacillus helveticus having high proteolysis activity

A technology of lactobacillus and lactic acid bacteria, applied in the field of lactic acid bacteria, can solve the problems of unstudied fermented milk, etc., and achieve the effect of good flavor, easy to drink or eat

Inactive Publication Date: 2013-07-24
ASAHI GRP HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no known case where fermented milk is directly used for the intake of functional amino acids and functional peptides

Method used

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  • Bacterium lactobacillus helveticus having high proteolysis activity
  • Bacterium lactobacillus helveticus having high proteolysis activity
  • Bacterium lactobacillus helveticus having high proteolysis activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0160] [Example 1] Identification of Lactobacillus helveticus CP3232 bacterial strain

[0161] Lactobacillus helveticus CM4 strain (accession number FERM BP-6060) was cultivated. A new Lactobacillus helveticus CP3232 strain showing properties as described in Examples 2 to 5 was obtained from a mutant strain derived from said CM4 strain.

[0162] The Lactobacillus helveticus CP3232 strain was anaerobically cultured in an MRS agar medium at 37° C. for 24 hours, and then the morphologically observed characteristics of the obtained colonies, their physiological characteristics and the like were examined. In addition, the 16S rRNA sequence was determined and confirmed to have 99.9% homology with another Lactobacillus helveticus strain (type strain). Therefore, it was revealed that the strain thus obtained definitely belongs to Lactobacillus helveticus and is a new strain.

Embodiment 2

[0163] [Example 2] Preparation of fermented milk

[0164]9.00% (w / w) reconstituted skim milk powder was sterilized by increasing the temperature to 95°C and then cooled to 15°C. The resultant was used as a milk medium. Fermented milk containing lactic acid bacteria (5%) was added to the milk medium (for inoculation), followed by culturing at 37° C. for 24 hours. Repeat this step, and use the fermented milk thus obtained as a starter. This starter (3%) was added to the milk medium, followed by culturing at 32°C or 37°C for 24 hours. The pH and acidity of this fermented milk were measured. Acidity was measured using a Hiranuma automatic titrator COMTITE-450 (Hiranuma Sangyo Co., Ltd.). Lactobacillus helveticus CP3232 strain and Lactobacillus helveticus CP3264 strain (isolated from a commercially available antihypertensive drug), Lactobacillus helveticus CM4 strain (accession number FERM BP-6060), Lactobacillus helveticus CP1092 strain as lactic acid bacteria were used , Lac...

Embodiment 3

[0171] [Example 3] Determination of free amino acids in fermented milk supernatant

[0172] The fermented milk obtained in Example 2 was centrifuged at 10000 g for 10 minutes. The supernatant was appropriately diluted with distilled water, the solution was filtered with a 0.2 μM membrane filter, and then measured by high-performance liquid chromatography (HPLC, Shimadzu Corporation) using an amino acid automatic analyzer (C-R7A / LC-10A). Analysis conditions are described below.

[0173] Column: Shim-pack Amino-Li (100mmL×6.0mmI.D.)

[0174] Ammonia trap: Shim-pack ISC-30Li (50mmL×4.0mmI.D.)

[0175] Column temperature: 38°C to 58°C

[0176] Reaction temperature: 65°C

[0177] Mobile phase: Shimadzu mobile phase kit for amino acid analysis, namely, Li type

[0178] Reaction solution: Shimadzu reaction solution kit for amino acid analysis, i.e., OPA reagent

[0179] Flow rate: 0.4ml / min.

[0180] Sodium hypochlorite was added to the reaction solution to detect proline. Fl...

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Abstract

The present invention provides a lactic acid bacterium which has high proteolysis activity and thus has excellent peptide production ability and / or amino acid production ability. The present invention also provides fermented milk which contains an amino acid and / or a peptide at high concentration and has a good flavor. The present invention specifically provides a lactic acid bacterium belonging to the genus Lactobacillus helveticus, which is characterized by having at least one of the following properties (a)-(d). (a) The lactic acid bacterium produces 25 [mu]mol / ml or more of a free amino acid by fermenting animal milk. (b) The lactic acid bacterium produces 8.5 mg / ml or more of a peptide by fermenting animal milk. (c) The lactic acid bacterium produces XP peptide and / or XPP peptide in a total amount of 260 [mu]g / ml or more by fermenting animal milk. (d) The amount of IPP peptide and VPP peptide per acidity of fermented milk that is obtained by fermenting animal milk at 30-34 DEG C (inclusive) is 6.0 mg / g or more in terms of VPP.

Description

technical field [0001] The present invention relates to a novel lactic acid bacterium belonging to Lactobacillus helveticus. The present invention also relates to fermented milk obtained by fermenting milk with the lactic acid bacteria, and a composition or food-drinks containing the lactic acid bacteria. The present invention further relates to a method for producing amino acids and / or peptides using said lactic acid bacteria. Background technique [0002] Milk proteins and food proteins are known to potentially contain functional peptides. Attempts have been made to use milk proteins or food proteins processed by enzymatic hydrolysis or fermentation. For example, as such functional peptides, tripeptides VPP and IPP have been reported to have angiotensin converting enzyme (ACE) inhibitory activity and thus exhibit hypotensive effects (Patent Documents 1-4 and Non-Patent Document 1). [0003] Dipeptide YP, which is a milk-derived functional peptide other than VPP and IPP,...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23C9/13C12P13/04C12P21/06C12N15/00C12R1/225
CPCC12N1/20A23C9/1234A23Y2220/39A23L1/3014C12P21/06A23L1/3053C12P13/04A23C9/127C12R1/225A23L33/135A23L33/18C12R2001/225C12N1/205A23V2400/147
Inventor 若井丈人畑中美咲中村康则山本直之
Owner ASAHI GRP HLDG LTD
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