Penicillium pinophilum strain and method for preparation of dextranase from the same

A technology of dextranase and Penicillium pinophilum, which is applied in the field of microbial technology and fermentation engineering, can solve the problems of high preparation cost and low dextranase activity, and achieve the effects of increasing fluidity, good thermal stability and reducing viscosity

Active Publication Date: 2015-09-16
SHANGHAI HUAMAO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the dextranase at home and abroad is mainly obtained from bacterial strains such as Penicillium lilacinum and Chaetomium sp., and because the prepared dextranase enzyme activity is low and the preparation cost is high, there is no A large number of reports on the application in the sugar industry

Method used

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  • Penicillium pinophilum strain and method for preparation of dextranase from the same
  • Penicillium pinophilum strain and method for preparation of dextranase from the same
  • Penicillium pinophilum strain and method for preparation of dextranase from the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A method for isolating dextranase-producing high-enzyme activity strain H6, comprising the steps of:

[0044] Add 1g of soil samples (soil samples were taken from Hefei, Anhui, Bengbu, Anhui, and Chengdu, Sichuan) to 99ml sterile water to prepare soil dilution, add penicillin solution (50μl to 100ml) into the PDA medium, pour it into a petri dish, and let it stand Let cool into a flat plate. Use the dilution coating method to apply the diluted soil solution on the plate respectively, and then place it upside down in a constant temperature incubator at 28°C for cultivation. After the bacteria grow out, use the inoculation needle to pick the strain into another sterile screening culture Base (Formulation: Dextran T20001g, NaNO 3 0.3g, K 2 HPO 4 ·3H 2 O 0.4g, MgSO 4 ·7H 2 O 0.02g, KCl 0.02g, FeSO 4 ·7H 2 (20.001g, agar 1.6g, water 100ml, pH3.0-3.5) carry out line separation, then the single bacterium colony that can grow on the screening medium is picked, and is tr...

Embodiment 2

[0049] The culture method of above-mentioned bacterial strain H6, it comprises the steps:

[0050] First use solid PDA medium, the solid PDA medium components are: every 100ml of water contains 20g of potatoes, 2g of sucrose, and 1.6g of agar; Activation culture, and then use liquid PDA medium for culture after 2 days. The components of liquid PDA medium are: 20 g of potatoes and 2 g of sucrose per 100 ml of water; ℃, 200r / min constant temperature culture for 7 days.

Embodiment 3

[0052] A kind of technology that utilizes microbial liquid fermentation of the present invention to prepare dextranase, it comprises the steps to carry out:

[0053] 1) Strain activation: using sterilized solid PDA medium, inoculate the H6 strain on the test tube medium, cultivate it at 28°C-30°C for 48-60h, and then use it to prepare the strain seed solution;

[0054] 2) Fermentation culture of bacterial strain seeds: the composition of the seed medium is to contain 1.5g of Dextran 7kDa per 100ml of water, KNO 3 2.5g, K 2 HPO 4 ·3H 2 O 0.1g, MgSO 4 ·7H 2 O 0.05g, FeSO 4 ·7H 2 O 0.001g, pH3.0-3.5, the bacterial strain in step 1) is inoculated in described seed culture medium, is placed on constant temperature shaker 32-34 ℃, cultivates 48-72h under rotating speed 200r / min as seed liquid, Then the seed solution is used for fermentation to prepare dextranase;

[0055] 3) Preparation of dextranase by fermentation: the components of the fermentation medium are 4.5g of Dext...

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Abstract

The invention provides a dextranase producing strain with high enzyme activity and a method for preparation of dextranase from the strain. The strain is characterized in that it is Penicillium pinophilum H6 separated from soil, the strain is preserved by China General Microbiological Culture Collection Center with a preservation number of CGMCC No.9260. The invention successfully utilizes Penicillium pinophilum H6 fermentation to prepare dextranase, also can produce dextranase with high enzyme activity, and the produced dextranase is extracellular enzyme. Preparation of dextranase by Penicillium pinophilum H6 fermentation process is characterized in that the most suitable action temperature is 40DEG-50DEG C, the most suitable pH is 5.0-6.0, and the stable pH is 3.0-10.0. The level of Penicillium pinophilum H6 fermentation for production of dextranase is 345U / ml.

Description

Technical field: [0001] The invention relates to the field of microbial technology and fermentation engineering, in particular to the separation of a dextranase-producing Penicillium pinophilum strain and a process for preparing dextranase with high enzyme activity by using the strain. Background technique: [0002] Dextran (Dextran) is a polymer formed by the dehydration of several glucans, which is mainly connected by glucose α-1,6 glycosidic bonds. It is produced by Leuconstoc mesenteriodes (Dextransucrase; E.C.2.4. 1.5) The synthesized dextran contains 95% α-1,6 glucosidic bonds, and contains a small amount of branched structures of other glycosidic bonds. According to the molecular weight, dextran can be divided into the following types: (1) micro-molecular dextran (dextran 10, average molecular weight below 10,000); (2) small molecule dextran (dextran 20, average molecular weight 10,000-25,000); (3) Low molecular dextran (dextran 40, average molecular weight 25,000-50...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/44C12R1/80
CPCC12N1/14C12N9/1051C12Y204/01005C12N1/145C12R2001/80
Inventor 张洪斌张宇琪李若菡胡雪芹
Owner SHANGHAI HUAMAO PHARMA
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