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A cellulase-producing mutant strain Fusarium yl2 and its screening and culture methods

A technology of cellulase and mutagenic strains, applied in the biological field, can solve the problems of low cellulase enzyme activity and low enzymatic hydrolysis efficiency, and achieve the goals of simplifying the screening process, wide application prospects, and good cellulase production ability Effect

Active Publication Date: 2018-02-16
INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cellulase is obtained through the fermentation of cellulase-producing bacteria, but the current problem is that the enzyme activity of cellulase is not high, and the enzymatic hydrolysis efficiency is low

Method used

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  • A cellulase-producing mutant strain Fusarium yl2 and its screening and culture methods
  • A cellulase-producing mutant strain Fusarium yl2 and its screening and culture methods
  • A cellulase-producing mutant strain Fusarium yl2 and its screening and culture methods

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Experimental program
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Embodiment 1

[0037] The present invention provides a cellulase-producing bacterium, and the classification of the cellulase-producing bacterium is named Fusarium YL2, Fusarium sp.YL2, Fusarium ATCC11599 purchased from the American Type Culture Collection ( Fusarium sp.ATCC11599) was a mutant Fusarium strain obtained by combined mutagenesis with UV and NTG. The present invention produces cellulase mutagenic strain Fusarium YL2 ( Fusariumsp.YL2) was deposited at the Chinese Type Culture Collection Center of Wuhan University in Wuhan, China on November 5, 2014, with the deposit number CCTCC M 2014551.

Embodiment 2

[0039] The screening method of the mutagenic strain Fusarium YL2 producing cellulase described in embodiment 1, its steps are:

[0040] (1) Activation of strains

[0041] The starting strain Fusarium ATCC 11599 ( Fusarium sp. ATCC 11599) was transferred to PDA slant medium, cultured at a constant temperature of 28-32°C for 3-4 days; the starting strain Fusarium ATCC 11599, Fusarium sp. ATCC 11599, purchased from the American Type Culture Collection ATCC;

[0042] Select the well-growing activated slant strains and transfer them to the Erlenmeyer flasks with seed medium, and shake them at 28-33°C for 36 hours at 200±20 rpm;

[0043] Draw the cultivated seed culture solution and centrifuge at 10000 rpm for 5 min, discard the supernatant;

[0044] Re-suspend the precipitate with normal saline and transfer it to a conical flask with glass beads, shake and mix well;

[0045] Remove the mycelium by filtering through a sterile funnel with gauze to obtain a spore suspension;

[...

Embodiment 3

[0061] The cultivation method of the mutagenic strain Fusarium YL2 producing cellulase described in embodiment 1, its steps are:

[0062] (1) Strain activation: the mutagenic strain Fusarium YL2 with high cellulase production was transferred to PDA slant medium for culture, and cultured at 30°C for 3-5 days;

[0063] (2) Seed culture: select a well-grown slant as a seed, and pick a piece (about 1cm) with an inoculation needle 3 ) into the seed medium, and cultured at 30°C for 3-4 days under shaking conditions of 200±20 rpm to obtain the seed solution;

[0064] (3) Fermentation culture: Inoculate the obtained seed culture solution in the fermentation medium with an inoculum amount of 4-8% (v / v), and culture at 28-32°C and 200±20 rpm for 7-9 days.

[0065] Fusarium YL2 ( Fusarium sp.) strain characteristics:

[0066] (1) Bacteria morphology: In the liquid culture, several bacteria are connected into a rope-like main hyphae, and gradually form a dead branch with the growth of...

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Abstract

The invention discloses a high-yield cellulase-producing Fusarium sp. YL2 (Fusarium sp.), which is preserved in the China Center for Type Culture Collection (CCTCC for short), with a preservation number of CCTCC M 2014551; fermentation medium used for cellulase-producing Fusarium sp. Composition: cellulose powder 40g / L, peptone 10g / L, KNO36g / L, ammonium sulfate 3g / L, KH2PO418 g / L, KCl1.6g / L, MgSO4·7H2O 1.2g / L, Tween-80 1.2 g / L , FeSO4 7H2O 0.01g / L, MnSO4 6H2O 0.01g / L, CuSO4 7H2O 0.01g / L, pH 4.0~5.0. The mutagenized strain Fusarium YL2 of the present invention can produce high cellulase in the fermentation medium, and its filter paper enzyme activity, carboxymethyl cellulose enzyme activity, microcrystalline cellulase activity are respectively 10.65 IU / mL, 101.38 IU / mL and 3.33 IU / mL. The cellulase high-yielding fungus Fusarium YL2 and the cellulase produced by the invention have broad application prospects in multiple industries such as biofuel, textile, resource regeneration, food fermentation, feed and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a mutagenic strain Fusarium YL2 with high cellulase production. The invention also relates to the screening and culturing method of the Fusarium YL2, which has better filter paper enzyme activity, carboxymethyl cellulose enzyme activity and microcrystalline cellulase activity. Background technique [0002] Cellulose is the main component of plants, accounting for about 33.4-50% of the dry weight of plants. It is estimated that nearly 100 billion tons of plant cellulose substances can be produced on average in the world every year. Cellulose is a linear straight-chain polymeric macromolecular polysaccharide composed of multiple glucose residues linked by β-1,4-glycosidic bonds, which can be hydrolyzed under various hydrolysis conditions (such as acid hydrolysis, alkali hydrolysis, enzymatic hydrolysis, etc.) Continuous bond breaking, depolymerization, and finally decompos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N13/00C12N15/01C12R1/645
CPCC12N1/14C12N13/00C12N15/01C12N1/145C12R2001/77
Inventor 袁林郭建军曾静傅筱冲邱小忠靳亮
Owner INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI
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