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Myceliophthora thermophilia strain and application thereof in aspect of producing keratinase

A technology of Myceliophthora thermophila and keratinase is applied in the field of Myceliophthora thermophila strains to achieve the effects of reducing the probability of contamination, growing rapidly and increasing the reaction temperature

Inactive Publication Date: 2012-07-25
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on keratinase produced by thermophilic fungi has not been reported yet.
Other fungi of the genus Myceliophthora have not been reported to produce keratinase

Method used

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  • Myceliophthora thermophilia strain and application thereof in aspect of producing keratinase
  • Myceliophthora thermophilia strain and application thereof in aspect of producing keratinase
  • Myceliophthora thermophilia strain and application thereof in aspect of producing keratinase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] (1) Source of material

[0015] Soil samples collected from Pingwu, Sichuan.

[0016] Martin's Medium: KH 2 PO 4 1g, MgSO 4 .7H 2 O 0.5g, peptone 5g, glucose 10g, agar 18g, Bengal red 1 / 300mL. Sterilization conditions: 121°C, 30min. Wait until the temperature is cooled to about 40°C before use, add penicillin 20 / mg, streptomycin 40μg / mg.

[0017] PDA medium: cut 200g of potatoes into pieces, add about 500mL of water, boil for 30min, filter, add 20g of glucose, 20g of agar, make up 1000mL of water, and the pH is natural. Sterilization conditions: 121°C, 30min.

[0018] (2) Isolation and purification of bacterial strains

[0019] Weigh 2g of soil sample into a conical flask filled with 20mL of sterile water and glass beads, shake for about 10min, so that the soil sample is evenly dispersed in the diluent to become a soil suspension.

[0020] Pipette 1mL of soil suspension into 9mL of sterile water, and dilute in turn by 10 times, usually at 10 -1 ~10 -3 .

[...

Embodiment 2

[0027] The method for Myceliophthora thermophila GZUIFR-H94.1 strain producing keratinase, comprises the following steps:

[0028] (1) culture medium

[0029] Shake flask fermentation medium: chicken feather powder 20.10g, urea 0.98g, NaCl 5g, K 2 HPO 4 6g, KH 2 PO 4 1g, water 1000m, initial pH is 7.9.

[0030] (2) Liquid fermentation culture

[0031] Bacterial strain of the present invention is inoculated in the petri dish containing solid PDA medium, cultivated for 3 days at 38°C, punched out with a hole puncher with a diameter of 7mm, and then inoculated the mycelium block into 250mL of 50mL re-screened liquid medium. In the triangular flasks, each bottle was connected with 3 wells of mycelium blocks, and each group was replicated 3 times, placed in a shaker at 38°C, kept at 140rpm for static culture, and the enzyme activity was measured.

[0032] (3) Preparation of crude enzyme solution

[0033] Preparation of spore liquid: take the strains stored in the refrigera...

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PUM

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Abstract

The invention discloses Myceliophthora thermophilia strain and an application thereof in the aspect of producing keratinase, the preservation number of Myceliophthora thermophilia strain GZUIFR-H94.1 is CCTCC NO: M 209140. The method for producing keratinase by the Myceliophthora thermophilia strain GZUIFR-H94.1 takes 20.10g / L of chicken feather powder as carbon source, nitrogen source and inductor, takes 0.98g / L of urea as supplement nitrogen source, and takes 5g / L of NaCl, 6g / L of K2HPO4 and 1g / L of KH2PO4 as enzyme-producing culture medium, wherein, the enzyme-producing temperature is 38 DEG C, the initial pH value is 7.9, and the above materials are continuously cultured for 6 days. The strain can grow fast, is thermophilic, and has stronger keratinase producing ability.

Description

technical field [0001] The present invention relates to Myceliophthora thermophila strain and application thereof. Background technique [0002] Keratinase (keratinase) is a highly specific protease that can specifically decompose keratin, and keratin exists widely in nature, mainly derived from animal ectoderm cells, including skin and skin derivatives, It is its structural protein, which can be divided into two types: α and β. This type of protein has a stable chemical structure and is a hard protein with strong resistance. Its structure is very stable through disulfide bonds, hydrogen bonds, salt bonds and other cross-bonds. It is insoluble in water and difficult to be digested by trypsin and stomach. Degraded by common proteases such as protease, but can be degraded by keratinase. Keratinase has the property of degrading natural keratin, belongs to the family of alkaline serine proteases or metalloproteases, and is considered a class of induced enzymes. Because kerati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/58C12R1/645
Inventor 梁建东韩燕峰梁宗琦张继伟杜文
Owner GUIZHOU UNIV
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