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Hypocrea lixii strain and method for preparing dextran enzyme with the Hypocrea lixii strain

A technology of dextranase and bacterial strains, which is applied in the field of microbial technology and fermentation engineering, can solve the problems of high production cost and low dextranase enzyme activity, and achieve the effects of promoting clean production, enhancing competitiveness, and improving product quality

Active Publication Date: 2011-10-19
SHANDONG JINYANG 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 the prepared dextranase enzyme activity is low and the preparation cost is high. A large number of reports on the application in the sugar industry

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A method for isolating strain F1002 producing dextranase high enzyme activity, which comprises the steps:

[0036] 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 (recipe: Dextran T2000 1g, 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 (0.001g, agar 1.6g, water 100ml, pH5.0~5.5) carry out streak separation, then the single bacterium colony that can grow on the screening medium is picked, and is...

Embodiment 2

[0041] The culture method of above-mentioned bacterial strain F1002, it may further comprise the steps:

[0042] First use solid PDA medium, the components of which are as follows: every 100ml of water contains 20g of potatoes, 2g of sucrose, and 1.6g of agar; Activation culture, after two days, culture in liquid PDA medium, liquid PDA medium components: every 100ml of water contains 20g of potatoes, 2g of sucrose; inoculate the strain activated on the slant of the test tube in the conical flask PDA liquid medium at 30°C , 220r / min constant temperature culture for 5 days.

Embodiment 3

[0044] A kind of technology that utilizes microbial liquid fermentation of the present invention to prepare dextranase, it comprises the following steps to carry out:

[0045] 1) Strain activation: use sterilized solid PDA medium to inoculate the F1002 strain on the test tube medium, cultivate it at 28-30°C for 48-60h, and then use it to prepare the strain seed solution;

[0046] 2) Fermentation culture of bacterial strain seeds: the components of the seed medium are 1g of Dextran 70kDa per 100ml of water, 0.5g of peptone, K 2 HPO 4 ·3H 2 O 0.4g, MgSO 4 ·7H 2 O 0.02g, KCl 0.02g, FeSO 4 ·7H 2 O 0.001g, pH 5.0~5.5, inoculate the bacterial strain in step 1) into the described seed medium, place on a constant temperature shaker at 28-30°C, and cultivate for 48-72 hours at a rotating speed of 220r / min as a seed solution , and then using the seed liquid for fermentation to prepare dextranase;

[0047] 3) Preparation of dextranase by fermentation: the components of the ferment...

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Abstract

The invention discloses a dextran enzyme producing strain with a high enzyme activity, and a method for preparing dextran enzyme with the strain. The strain is characterized by being a Hypocrea lixii F1002 strain separated from soil and being preserved by the Chinese General Microbiological Culture Collection Center, wherein the preservation number of the strain is CGMCC No. 4725. In the invention, dextran enzyme is prepared successfully by Hypocrea lixii F1002 fermentation, and dextran enzyme prepared by the method has a high enzyme activity and is an extracellular enzyme. The dextran enzymeprepared by the method is characterized in that an optimal action temperature is in a range of 25 to 30 DEG C; an optimal PH is in a range of 5.0 to 5.5; and a stabilization PH is in a range of 4.0 to 6.5. A degree of producing dextran enzyme by Hypocrea lixii F1002 fermentation is greater than 60 U / 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 Trichoderma harzianum strain and a process for fermenting and preparing dextranase with high enzyme activity by using the strain. Background technique: [0002] In the pharmaceutical industry, dextran with a molecular weight of 70kDa, 40kDa, and 20kDa is currently recognized as an excellent plasma substitute, which can increase blood volume and improve microcirculation, and is mainly used clinically to treat hemorrhagic shock. At present, more than 90% of dextran manufacturers in my country mostly use hydrochloric acid to hydrolyze high-molecular dextran. First, high-molecular dextran is synthesized by dextran sucrase, and then hydrolyzed by hydrochloric acid to obtain middle-molecular and small-molecular pharmaceutical grade dextran. Because this process uses hydrochloric acid to hydrolyze to produce pharm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/46C12R1/885
Inventor 张洪斌吴定涛胡雪芹黄丽君
Owner SHANDONG JINYANG PHARMA
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