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Culture medium and method for producing chitinase by Lecanicillium

A technology of chitinase and culture medium, which is applied in the field of culture medium for the production of chitinase by the fermentation of the lecanococcus fungus. The effect of high yield, short fermentation cycle and mild conditions

Inactive Publication Date: 2013-03-27
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the insect pathogenic fungi such as Coccus laxella, Ascospora, Paecilomyces, Metarhizium anisopliae and Beauveria bassiana have been studied more. The most extensive (Xiao Yan et al., 2001), but the research on the chitinase produced by L.

Method used

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  • Culture medium and method for producing chitinase by Lecanicillium
  • Culture medium and method for producing chitinase by Lecanicillium
  • Culture medium and method for producing chitinase by Lecanicillium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Single factor experiment to determine the optimal composition of culture medium

[0020] Implementation steps:

[0021] (1) Preparation of seed culture and basal medium: 1L of 20% potato juice (boiled from potato slices), 20g of glucose, natural pH. Packed in 250mL Erlenmeyer flask, each Erlenmeyer flask contains 150mL. 1.01MPa, sterilize for 20min. Antibiotics can be added to pre-treat the medium before use. Basal medium (liquid induction medium): powdered chitin 2%, KH 2 PO 4 0.1%, MgSO 4 。 7H 2 O 0.05%, NaCl 0.02%, pH6.5.

[0022] (2) Production of fermented seeds: Inoculate Lecanococcus sp. FJ28 on potato agar medium, culture at 26°C for 5 days, and activate after it produces spores; inoculate the activated strains with spore powder to the seeds with an inoculation needle The culture medium was cultured at 26°C for 4 days at a rotational speed of 160 r / min, which was the fermentation broth.

[0023] (3) Preparation of enzyme liquid: take the fermentat...

Embodiment 2

[0038] Orthogonal experiment to determine optimal fermentation conditions

[0039] (1) Orthogonal experiment to determine the best medium

[0040]On the basis of the culture medium determined by the single-factor experiment, various components and pH values ​​were changed to configure different culture medium, and the method was the same as steps (1) and (2) in Example 1. See Table 1. Inoculate 250mL of the same amount of inoculum (10%) into 250mL of culture solution, and culture at 26°C with a rotation speed of 160 r / min for 4 days. Then measure the enzyme activity according to the steps (3) and (4) of Example 1. Select the experimental group number with the optimal result of the orthogonal experiment and the highest enzyme production, and carry out the verification experiment. In order to avoid being cumbersome, the preparation of the culture medium, the preparation of the crude enzyme solution, and the enzyme activity assay method in the embodiment are all the same as...

Embodiment 3

[0057] Fermentation method of Chitinase produced by Lecanococcus cerevisiae FJ28

[0058] (1) Preparation of fermented seeds

[0059] (A) Strain and culture medium

[0060] Bacteria: Lexococcus FJ28

[0061] Slant medium: PDA medium

[0062] Liquid seed medium: 1L of 20% potato juice, 20g of glucose, natural pH, 1.01MPa, sterilized for 20min.

[0063] (B) Preparation of seed solution

[0064] Transfer the pure species of Lecanococcus spp. FJ28 on the slant to several 250mL Erlenmeyer flasks filled with 100mL of culture medium, and then culture them at 26°C for 11d at the speed of 160 r / min on a reciprocating shaker until the hyphae grow. Robust, when the bacterial liquid is viscous, it means that the seeds have grown well.

[0065] (2) Fermentation culture

[0066] Fermentation medium: optimize the results of orthogonal experiments according to the medium, that is, 1% (w / v) maltose, 1% (w / v) yeast extract, 4×10 -4 mol / L Ca 2+ , 0.01%V B6 , pH 5.5, prepared. ...

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Abstract

The invention relates to a fungal fermentation culture medium and process. More particularly, the invention relates to a culture medium and method for producing chitinase by Lecanicillium fermentation. The formula of the culture medium comprises the following raw materials: 5-15g / L of fructose or maltose, 5-15g / L of yeast extract powder or yeast extract extractum, 2-8*10<-4>mol / L of K<+> or Ca<2+>, 0.05-0.2g / L of Vc or VB6, and the balance of water; and the initial pH of the culture medium is 5.5-7.0. According to the method, a fermented seed solution is inoculated to the culture medium, the liquid containing quantity of a 250 mL triangular flask is 50 mL, the inoculum size is 15mL, and chitinase is cultured at 26 DEG C at the rotational speed of 160 r / min. The fermentation method is short in fermentation period, mild in condition, and easy to control; and the culture medium provided for producing chitinase by Lecanicillium fermentation has the advantages that the chitinase yield is high, the chitinase activity is high, and the like.

Description

technical field [0001] The invention relates to a fungal fermentation medium and a process. More specifically, the present invention relates to a kind of lecanococcus ( Lecanicillium ) culture medium and method for producing chitinase by fermentation. Background technique [0002] Lexococcus ( Lecanicillium ) belongs to the hyphomycetes (Hyphomycetes) , widely distributed in tropical, subtropical and temperate regions, can parasitize scales, aphids, mites and whiteflies, and can also parasitize some pests of Lepidoptera, nematodes and thrips. Since the 1970s, some European countries, the United States, Japan and other countries have paid great attention to the use of Lecanococcus to control greenhouse vegetable pests. At present, the insect pathogenic fungi such as Coccus laxella, Ascospora, Paecilomyces, Metarhizium anisopliae and Beauveria bassiana have been studied more. The most extensive (Xiao Yan et al., 2001), but the research on the chitinase produced by L. lec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12R1/645
Inventor 邱君志苏礼超蔡志雄涂洁宋飞飞邱云锋李小霞何肖云毛丽慧谢小聪
Owner FUJIAN AGRI & FORESTRY UNIV
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