Paecilomyces thermophila mutant strain and mutation induction method and application thereof

A technique for mutagenizing bacterial strains, Paecilomyces, applied in the field of microorganisms

Inactive Publication Date: 2017-09-26
XUZHOU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the production of gluco

Method used

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  • Paecilomyces thermophila mutant strain and mutation induction method and application thereof
  • Paecilomyces thermophila mutant strain and mutation induction method and application thereof
  • Paecilomyces thermophila mutant strain and mutation induction method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Example 1 Screening of the original strain Paecilomyces thermophilia producing heat-resistant and acid-resistant glucose oxidase

[0043] This embodiment illustrates the screening method of the original bacterial strain Paecilomyces thermophila for producing heat-resistant and acid-resistant glucose oxidase, and the specific steps are as follows:

[0044] Step 1 Sampling and sample processing:

[0045] Using the multi-point mixed soil sample collection method, soil samples were taken from the compost in the countryside of Yunlong District, Xuzhou City, Jiangsu Province, which was undergoing high-temperature fermentation, and the sampling depth was 15-20cm; The depth and quality are uniform. Take 500g of each mixed sample. The sampling season is summer. Weigh 5g of solid soil samples, put them into a triangular flask filled with glass beads and 50mL of sterilized saline, shake at 180r / min for 30min, and prepare to form a suspension, and then put the triangular flask int...

Embodiment 2

[0068] Example 2 Mutation Breeding Using Paecilomyces thermophila Starting Strains

[0069] The original strain XZ36 obtained by screening was used as the starting strain for further compound mutagenesis. Mutagenesis procedure such as figure 1 shown. Include the following steps:

[0070] 1. Put the Paecilomyces thermophila XZ36 screened in Example 1 as the starting strain for mutagenesis to the slant of solid nutrient activation medium, and culture it at 28-34° C. for 3-4 days. The solid nutrient activation medium is: 800ml of potato extract, 20g of sucrose, 5-8g of yeast powder, NaNO 3 0.2g, K 2 HPO 4 0.1, KCl 0.05, MgSO 4 0.05g, agar powder 15g, add deionized water to 1000ml, pH4.0-5.0.

[0071] 2. Scrape the spores from the activated slant and inoculate them into the shake flask culture medium, culture at 28-34°C, 200r / min for 28-32h, in which the shake flask culture medium is: 800ml of potato extract, 20g of sucrose, 5-8g of yeast powder, NaNO 3 0.2g, K 2 HPO...

Embodiment 3

[0102] Example 3 Application of Glucose Oxidase Production in Paecilomyces thermophila Mutagenic Strain XZPT6 in Weaned Piglets

[0103] Select 60 healthy, weaned piglets with a body weight of about 8kg, and divide them into 2 groups with 30 piglets in each group according to the principle of similar body weight and half male and female piglets. Wherein the control group is the basic dietary raw material without adding glucose oxidase, and the test group is to add the glucose oxidase produced by Paecilomyces thermophila mutagenic strain XZPT6 in the basic dietary raw material, and the addition method is as follows: the glucose oxidase is fermented The solution was centrifuged at 8000r / min for 20min, and the supernatant was taken to obtain the glucose oxidase crude enzyme solution. Before feeding piglets, the glucose oxidase crude enzyme solution is diluted 10 times and evenly sprayed into the solid feed, and 1-3ml of the diluted glucose oxidase crude enzyme solution is sprayed...

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Abstract

The invention discloses a Paecilomyces thermophila mutant strain and a mutation induction method and application thereof. The mutation induction method includes: Paecilomyces thermophila screened from high-temperature-fermented compost and used for producing high-temperature-resistant and acid-resistant glucose oxidase is used as the original strain, protoplast ultraviolet and ethyl methane sulfonate compound mutation induction is performed, and the screened strain is subjected to budding spore diethyl sulfate-microwave compound mutation induction to obtain the Paecilomyces thermophila mutant strain. The Paecilomyces thermophila mutant strain is preserved in China General Microbiological Culture Collection Center on January 11th, 2016, and the preservation number of the mutant strain is CGMCC No. 13182. Compared with a conventional mutation induction method, the mutation induction method has the advantages that the method is high in efficiency and simple in fast in screening, the productivity of the mutant strain is high, the yield of the mutant stain is 277.65% of that of the Paecilomyces thermophila original strain, fermentation liquor enzyme activity reaches 185U/ml, production cost is lowered, and the glucose oxidase produced by the mutant strain is good in high-temperature resistance and acid resistance and can be used as feed enzyme applied to feed industry.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to Paecilomyces thermophila with high yield of high-temperature-resistant and acid-resistant glucose oxidase and a screening method thereof. Background technique [0002] Glucose oxidase (GOD for short) is a very important enzyme in the field of enzyme technology. It can use molecular oxygen as an electron acceptor to exclusively catalyze the oxidation of β-D-glucose into gluconic acid and hydrogen peroxide. Because of its close relationship with the vital substances glucose and oxygen, it has been widely used in scientific research, medicine and food industry production. In the food industry, glucose oxidase can be used to remove glucose in food or medicine, thereby preventing browning of food or obtaining oligosaccharides with higher purity; it can remove oxygen from food to improve food quality and prolong food The purpose of shelf life; at the same time, glucose ox...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N15/01C12N13/00C12N9/04A23K20/189A23K50/30C12R1/79
CPCC12N1/14C12N9/0006C12N13/00C12N15/01C12Y101/03004C12N1/145C12R2001/79
Inventor 高兆建张铁柱李同祥刘恩岐曹建冬顾强沈彬彬
Owner XUZHOU UNIV OF TECH
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