Phospholipase C, food enzyme agent comprising same and processing method for food raw materials

A phospholipase and food technology, applied in the field of phospholipase C, can solve problems such as insufficient efficacy and safety problems

Inactive Publication Date: 2011-09-07
MITSUBISHI KAGAKU FOODS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0037] In this way, the following problems of phospholipase C can be listed: the conventional phospholipase C has insufficient efficacy as an enzyme, or has problems in safety, etc., so there is no enzyme agent containing phospholipase C that can be circulated in the market so far.

Method used

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  • Phospholipase C, food enzyme agent comprising same and processing method for food raw materials
  • Phospholipase C, food enzyme agent comprising same and processing method for food raw materials
  • Phospholipase C, food enzyme agent comprising same and processing method for food raw materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0183] Example 1: Phospholipase C was purified from the FERM ABP-10200 strain of Aspergillus oryzae

[0184] 1) Preparation of crude enzyme solution

[0185] In the 500ml Erlenmeyer flask (seed flask) that added 100ml of sterilized culture medium with the composition shown in Table 2, inoculate 1ml of 5% sodium deoxycholate solution through filter sterilization, and aspergillus oryzae (Aspergillus oryzae) The bacterial body of FERM ABP-10200 strain was shaken at 170rpm at 26°C for 7 days.

[0186] Table 2

[0187] culture medium

[0188]

[0189] Polypeptone 40g

[0190] Yeast Extract 5g

[0191] Dipotassium hydrogen phosphate 0.2g

[0192] Magnesium sulfate 0.5g

[0193] 1000ml after adding pure water

[0194]

[0195] After the cultivation, centrifugation was performed at 10000G for ten minutes at 4°C, and the obtained supernatant was used as a crude enz...

Embodiment 2

[0212] Example 2: Purification of Phospholipase C from IAM 13907 Strain of Aspergillus tamarii

[0213] 1) Preparation of crude enzyme solution

[0214] In the 500ml capacity Erlenmeyer flask (seed flask) that has the substratum of composition shown in 100ml Table 2, inoculate 1ml of the IAM 13907 strain of 5% sodium deoxycholate solution and Aspergillus tamarii through filter sterilization Bacteria were cultured with shaking at 170 rpm for 5 days at 26°C. After the incubation, centrifugation was performed at 10000G for 10 minutes at 4°C, and the obtained supernatant was used as a crude enzyme solution.

[0215] 2) Preparation of purified enzyme solution

[0216]In this example, purification was carried out in the same manner as described in 3) of Example 1 to obtain a purified enzyme solution.

[0217] 3) Molecular weight determination of purified enzyme

[0218] This example is to measure with the same method described in 4) in the embodiment 1. The purified enzyme show...

Embodiment 3

[0219] Example 3: Phospholipase C was purified from NBRC 4190 strain of Aspergillus oryzae

[0220] 1) Preparation of crude enzyme solution

[0221] Add 100ml of sterilized 500ml Erlenmeyer flask (seed flask) with the medium shown in Table 3, inoculate the bacterium of NBRC 4190 strain of Aspergillus oryzae, and shake at 170rpm at 26°C Cultured for 7 days.

[0222] table 3

[0223] culture medium

[0224]

[0225] Fish meal 50g

[0226] 1000ml after adding pure water

[0227]

[0228] After the incubation, centrifugation was performed at 10000G for 10 minutes at 4°C, and the obtained supernatant was used as a crude enzyme solution.

[0229] 2) Preparation of purified enzyme solution

[0230] First, 600 ml of the crude enzyme solution obtained in 1) was dialyzed 5 times every 12 hours using 8000 ml of 10 mM tris-hydrochloric acid buffer solution (pH 7.5). T...

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Abstract

The invention relates to phospholipase C, a food enzyme agent comprising the same and a processing method for food raw materials, providing the phospholipase C. The phospholipase C has the capability of efficiently hydrolyzing various phospholipids close to an acidic pH region or a neutral pH region, also has activity in a citric acid buffer solution, has thermal stability to a certain extent and also has the property of not hydrolyzing organic phosphate not containing a lipid part. The phospholipase C shows activity in the region with pH from acidic pH to neutral pH and does not substantially hydrolyze the organic phosphate except for the phospholipids.

Description

[0001] This application is a divisional application with the application number 200680053785.2 (international application number: PCT / JP2006 / 304710) filed by the applicant Mitsubishi Chemical Foods Co., Ltd. on March 10, 2006. technical field [0002] The present invention relates to a phospholipase C, a filamentous bacterium capable of producing the phospholipase C, a method for isolating the phospholipase C from a culture of the filamentous bacterium, a DNA encoding the phospholipase C, and the phospholipase C manufacturing method, etc. Specifically, the present invention relates to a phospholipase C that is particularly suitable for the food industry and the pharmaceutical industry, especially the phospholipase C produced by Aspergillus oryzae or Aspergillus tamarii in filamentous fungi , a filamentous bacterium producing the phospholipase C, a method for isolating and purifying the phospholipase C from a culture of the filamentous fungus, a DNA encoding the phospholipase C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/16A23L1/03C12R1/66C12R1/69A23L29/00
Inventor 长崎咏子深泽彻也小野泰典
Owner MITSUBISHI KAGAKU FOODS
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