Fusarium sp. BYB2 and application thereof for preparing acacia enzyme by microbial fermentation

A technology of BYB2 and gum arabic, which is applied in the field of gum arabic enzyme producing strains, can solve the problems of complex production process, low purity, high production cost, etc., and achieve the effects of simple nutritional requirements, high product yield and short action time

Active Publication Date: 2011-03-30
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention provides an arabic gumase-producing strain—Fusarium sp. BYB2, and its application in the preparation of L-arabinose, which overcomes the complex production process of L-arabinose and the low purity. Low, high production costs and other disadvantages

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Enrichment, isolation and screening of microorganisms producing arabinase

[0032] Add 45mL shaking flask screening medium to a 250mL Erlenmeyer flask, add 5g soil sample after sterilization, and culture at 30°C and 200r / min shaking conditions for 5 days, if the volume of the medium decreases due to water evaporation during the cultivation process , add sterile water to the original volume. Take 5 mL of the above-mentioned culture solution and inoculate it into another sterilized Erlenmeyer flask containing 45 mL of screening medium, and repeat the above-mentioned culture process, so that the number of microorganisms that can grow with gum arabic as the only carbon source increases.

[0033] Dilute the enriched culture solution with sterile water and spread it on the plate medium, and cultivate the culture plate in a biochemical incubator at 30°C until the number of colonies no longer increases. Observe the colonies on the plate, pick colonies with different...

Embodiment 2

[0038] Embodiment 2: Mutation breeding of high-yielding arabic gumase strain

[0039] The fusarium strain ZHB-306 screened in Example 1 was mutated by ultraviolet irradiation, and the mutant strains with improved activity of producing arabinase were screened.

[0040] Ultraviolet radiation mutagenesis method: After activating the strain ZHB-306 slant, scrape the spores into a conical flask containing 50mL of sterile water, shake at room temperature for 20-30min, and make a spore suspension. Count the spores with a hemocytometer under a microscope, and control the number of spores to 1×10 8 A / mL or so. Under red light, take 2 mL of the above-mentioned spore suspension and a sterile paper clip into 6 petri dishes with a diameter of 6 cm, place the petri dishes on a magnetic stirrer, and irradiate them under a preheated ultraviolet lamp for about 20 minutes for 5 ~10min. Take 0.5mL of the spore liquid after the above-mentioned irradiation treatment, make appropriate dilutions,...

Embodiment 3

[0044] Embodiment 3: the method for preparing arabic gum enzyme by Fusarium BYB2 fermentation

[0045] Using Fusarium BYB2 (CCTCC NO: M 2010087) as the strain, after optimizing the seed preparation method, enzyme production medium composition and fermentation conditions, the preparation method of arabic gum enzyme is as follows:

[0046] (1) Inoculate the strain of Fusarium BYB2 that was freeze-dried or preserved on the slant of the test tube into the slant medium, and the slant was cultured in a biochemical incubator at 30°C for 3 days. The final concentration of the slant medium consists of: potatoes 200g / L (peeled potatoes, cut into small pieces and add water, boil for 30min, filter with gauze to remove slag), sucrose 20g / L, agar 20g / L, solvent is water, pH 5-6.

[0047] (2) Use an inoculation loop to pick the Fusarium BYB2 spores after step (1) activated culture into the liquid seed medium, and the liquid seed culture is based on 30°C and 200r / min shaking conditions for 1...

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PUM

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Abstract

The invention provides a new acacia enzyme generating strain (Fusarium sp. BYB2) and an application thereof for preparing L-arabinose by fermentation. The Fusarium sp. BYB2 is preserved in the China Center for Type Culture Collection (CCTCC), the preservation number is CCTCC No: M 2010087, and the preservation date is April 19, 2010. The invention has the main advantages that (1) the Fusarium sp. BYB2 of the invention has simple nutritional requirements and can be cultivated easily; (2) the acacia enzyme generated by the Fusarium sp. BYB2 is exoenzyme which can be obtained without cell breakage; (3) the acacia enzyme generated by the Fusarium sp. BYB2 has high activity, and the activity of the crude enzyme liquid which is not separated and purified can reach 231.6U / mL under the preferable fermentation conditions; and (4) the acacia enzyme generated by the Fusarium sp. BYB2 has mild reaction conditions for hydrolyzing acacia, shorter action time and high product yield.

Description

(1) Technical field [0001] The invention relates to a new arabinase-producing strain—Fusarium sp. BYB2, and its application in the preparation of L-arabinose by microbial fermentation. (2) Background technology [0002] L-arabinose belongs to the five-carbon aldose, and it mostly exists in the form of arabinan, arabinoxylan, arabinose galactose and hemifibers of higher plants. L-arabinose is a low-calorie sweetener and an important pharmaceutical intermediate, and it can be used in the preparation of bacterial culture medium in the field of biochemistry and the synthesis of spices, etc., with a wide range of applications. However, due to the complex production process and high production cost of L-arabinose, the price remains high, which hinders its application in the fields of food or medicine. [0003] At present, the preparation methods of L-arabinose include chemical synthesis and acid hydrolysis. The chemical synthesis method has many steps, complex process and expens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/24C12P19/02C12R1/77
Inventor 应国清梅建凤廖青青陈建澍王鸿易喻
Owner ZHEJIANG UNIV OF TECH
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