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Method for transporting microbial cells

A technology of microbial bacteria and bacteria, which is applied in the field of transportation of microbial bacteria, can solve the problems of reduction, loss of microbial enzyme catalytic ability, etc., and achieve the effect of cooling cost reduction

Active Publication Date: 2014-03-05
MITSUBISHI CHEM CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

That is, the activity of the enzyme must be maintained to avoid the loss or reduction of the catalytic ability of the microbial enzyme due to the mixing, corruption, or lysis of bacteria during storage or transportation

Method used

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  • Method for transporting microbial cells
  • Method for transporting microbial cells
  • Method for transporting microbial cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 and 2、 comparative example 1

[0038] to cultivate

[0039] Rhodococcus rhodochrous J-1 strain (Rhodococcus rhodochrous J-1: FERM BP-1478) having nitrile hydratase activity was prepared using a mixture containing 2% by mass of glucose, 1% by mass of urea, 0.5% by mass of peptone, 0.3% of yeast extract, chlorine Aerobic culture was carried out in a medium (pH 7.0) with 0.05% by mass of cobalt chloride.

[0040] Preparation of bacterial cell suspension for preservation

[0041] The cultured microbial cells were recovered by centrifugation (12000 rpm, 20 minutes), and washed with a 0.1% sodium acrylate aqueous solution (pH 7.0). The above-mentioned aqueous solution was added to the washed bacterial cells to obtain a bacterial cell suspension (10% by mass as dry bacterial cells).

[0042] Preservation of bacterial suspension

[0043] Add 115mL (Example 1) and 70mL (Comparative Example 1) of the bacterial cell suspension prepared by the above method into a polyethylene container with a total volume of 120mL, ...

Embodiment 3

[0048] Embodiment 3, comparative example 2

[0049] Preparation of transformant with nitrile hydratase derived from Rhodococcus rhodochrous strain M8

[0050] (1) Preparation of chromosomal DNA of Rhodococcus rhodochrous M8 strain (hereinafter referred to as M8 strain.)

[0051] The M8 strain (SU1731814) can be obtained from the Russian strain center IBFM (VKPM S-926).

[0052] M8 strain was dissolved in 100mL of MYK (0.5% polypeptone, 0.3% Bacto yeast extract, 0.3% Bacto malt extract, 0.2% K 2 HPO 4 , 0.2% KH 2 PO 4 ) medium (pH 7.0), cultured with shaking at 30°C for 72 hours. Centrifuge the culture medium, and suspend the collected bacteria in 4 mL of Saline-EDTA solution (0.1M EDTA, 0.15M NaCl (pH8.0)). 8 mg of lysozyme was added to the suspension, shaken at 37°C for 1 to 2 hours, and then frozen at -20°C.

[0053] Next, 10 mL of a Tris-SDS solution (1% SDS, 0.1M NaCl, 0.1M Tris-HCl (pH 9.0)) was added to the suspension while shaking it steadily. Further, prote...

Embodiment 4、 comparative example 3

[0090] to cultivate

[0091] Pseudomonas marginalis DSM16275 strain (Pseudomonas marginalis DSM16275) having nitrile hydratase activity was prepared using a mixture containing 0.2% by mass of glycerin, 0.05% by mass of citric acid, 0.15% of yeast extract, 0.3% by mass of potassium dihydrogen phosphate, and dipotassium hydrogen phosphate Aerobic culture was carried out in a medium (pH 7.0) containing 0.7% by mass, 0.0004% by mass of iron sulfate, 0.01% by mass of manganese sulfate, 1.25% by mass of methylurea, and 0.001% by mass of cobalt chloride.

[0092] Preparation of bacterial cell suspension for preservation

[0093] The cultured microbial cells were recovered by centrifugation (12000 rpm, 20 minutes), and washed with a 0.1% sodium acrylate aqueous solution (pH 7.0). The above-mentioned aqueous solution was added to the washed bacterial cells to obtain a bacterial cell suspension (10% by mass as dry bacterial cells).

[0094] 100 mL (Example 4) and 80 mL (Comparative Ex...

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Abstract

A method for storing microbial cells having nitrile hydratase activity in a hermetically sealable container, and a method for transporting microbial cells having nitrile hydratase using the container are provided. The methods are characterized in that the vapor phase portion within the container is set to 2-20% (inclusive) of the total interior volume of the container, said vapor phase portion being filled with a suspension that contains the microbial cells.

Description

technical field [0001] The present invention relates to a kind of method, it is when the microbial thallus cell that has nitrile hydratase activity is filled in airtight container and is transported, by making the gas phase part inside the airtight container account for 20% or less of the total volume, thereby make nitrile hydratase The activity is maintained in a stable state for transport. Background technique [0002] Enzymes produced by microorganisms are used in various situations as catalysts for chemical transformation reactions. In particular, amides, carboxylic acids, α-hydroxycarboxylic acids, etc. which are important in the chemical industry can be produced inexpensively by using nitrile hydratase, nitrilase, etc., which have the ability to hydrate or hydrolyze nitrile groups. Furthermore, by using the above-mentioned enzymes having optically specific hydration or optically specific hydrolysis ability, it becomes possible to produce optically active carboxylic ac...

Claims

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Application Information

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IPC IPC(8): C12N9/78C12R1/01
CPCC12Y402/01084C12N9/88C12Y305/05001C12N9/96C12N9/78C12N1/04
Inventor 竹内雅人渡边文昭
Owner MITSUBISHI CHEM CORP
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