Application of Penicillium oxalicum EU2101 in preparation of cellulase preparation and degradation of cellulose

A EU2101, a technology for degrading cellulose, applied in the direction of enzymes, enzymes, hydrolase, etc., can solve problems such as large gaps in industrial applications

Active Publication Date: 2016-10-26
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cellulase activity and yield of Penicillium strains are far from the large-scale industrial application.

Method used

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  • Application of Penicillium oxalicum EU2101 in preparation of cellulase preparation and degradation of cellulose
  • Application of Penicillium oxalicum EU2101 in preparation of cellulase preparation and degradation of cellulose
  • Application of Penicillium oxalicum EU2101 in preparation of cellulase preparation and degradation of cellulose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0156] Embodiment 1, the identification of Penicillium oxalicum EU2101

[0157] Penicillium oxalicum EU2101 is among the original fungal strains in the laboratory of the inventor of the present application. After mutagenesis, the strain with higher cellulase-producing activity was obtained through screening. Using Penicillium oxalicum wild-type strain HP7-1 (CGMCC No.10781) as the starting strain, three rounds of Co 60 - Gamma ray irradiation, obtained a mutant strain TCO7-4 whose filter paper enzyme activity was higher than that of the original strain HP7-1. Then TCO7-4 and its mutant strain EU122 were subjected to combined mutagenesis and screening with ethyl methylsulfonate (EMS) and ultraviolet (ultraviolet, UV) to finally obtain the mutant strain EU2101.

[0158] The specific operation is to carry out Co 60 When γ-rays are irradiated, the spore suspension with a lethal rate of about 85% is selected for large-scale isolation and screening of the mutant strains; while the...

Embodiment 2

[0160] Example 2, Optimization of Cellulase Production Conditions by Penicillium oxalicum EU2101 Solid-state Fermentation

[0161] The experiment was repeated three times, and the specific steps of each repeated experiment were as follows:

[0162] 1. Preparation of spore suspension

[0163] 1. Sterilize the PDA solid medium at 115°C for 30 minutes to obtain a PDA plate.

[0164] 2. Wash the Penicillium oxalicum EU2101 spores cultivated on the PDA plate for 5 days with 0.1% (v / v, volume percentage concentration) Tween 80 aqueous solution, and the preparation concentration is 1×10 8 per mL of spore suspension.

[0165] 2. Optimization of the initial pH of the medium

[0166] 1. Preparation of fermentation media with different pH

[0167] Preparation of basic medium: the basic medium is composed of wheat bran and salt solution, wherein the proportion of wheat bran and salt solution is 10.0g:20mL. Salt solution consists of water, KH 2 PO 4 , (NH 4 ) 2 SO 4 , MgSO 4 ·7H...

Embodiment 3

[0230] Embodiment 3, utilize Penicillium oxalicum EU2101 to prepare cellulase preparation

[0231] 1. Preparation of fermentation medium

[0232] Prepare the optimal culture medium in Step 9 of Example 2.

[0233] 2. Preparation of spore solution

[0234] (1) Sterilize the PDA solid medium at 115° C. for 30 minutes to obtain a PDA plate.

[0235] (2) Penicillium oxalicum EU2101 was inoculated on a PDA solid plate, and placed in a constant temperature incubator at 28° C. for 5 days. The Penicillium oxalicum EU2101 spores cultivated on the PDA plate for 5 days were washed with 0.1% (volume percentage concentration) Tween 80 aqueous solution, and transferred to a 50 mL centrifuge tube containing several glass beads. Under the microscope, count the hemocytometer and adjust the spore concentration to 10 8 cells / mL, inoculated into the optimal medium above at an inoculum size of 10% (v / w).

[0236] 3. Preparation of cellulase preparation

[0237] (1) The fermentation medium in...

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Abstract

The present invention discloses Penicillium oxalicum EU2101 and applications of the Penicillium oxalicum EU2101 in preparation of a cellulase preparation and degradation of cellulose, wherein the Penicillium oxalicum EU2101 has the preservation number of CGMCC No.12769. The invention further provides a cellulase preparation prepared by using the Penicillium oxalicum EU2101, wherein the preparation method comprises: culturing the Penicillium oxalicum EU2101 at a temperature of 28-42 DEG C by using a culture medium, and collecting the fermentation product to obtain the cellulase preparation. According to the present invention, the experiment results prove that after the Penicillium oxalicum EU2101 is subjected to solid state fermentation culture, the extraction liquid has cellulase activities such as filter paper enzyme activity, CMC enzyme activity, Avicel enzyme activity, pNPC enzyme activity and pNPG enzyme activity, and can effectively hydrolyze the manihot esculenta residue being subjected to an alkali pretreatment to produce glucose, such that the application potential is provided in conversion and utilization of the manihot esculenta residue.

Description

technical field [0001] The invention relates to Penicillium oxalicum EU2101 in the field of biotechnology and its application in preparing cellulase preparations and degrading cellulose. Background technique [0002] With the continuous increase of population, the rapid consumption of non-renewable fossil energy such as coal and oil, and the severe environmental problems brought about by the use of fossil energy, people gradually realize that they are looking for a renewable and environmentally friendly new energy. importance. Cellulose-derived fuel ethanol has a higher octane number than gasoline, resulting in less exhaust emissions, making fuel ethanol widely considered as a substitute for or an additive in fossil energy. [0003] Lignocellulosic substances are the most abundant and widely distributed renewable biological resources in the world (Kuhad, R.C., Singh, A., Eriksson, K.E.L. 1997. Microorganisms and enzymes involved in the degradation of plant fiber cell walls[...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12P19/14C12P19/12C12P19/02C12P7/10C12R1/80
CPCC12N1/14C12N9/2437C12N9/2445C12P7/10C12P19/02C12P19/12C12P19/14C12Y302/01004C12Y302/01021C12Y302/01091C12N1/145C12R2001/80Y02E50/10
Inventor 冯家勋赵帅苏临辉蒋随新
Owner GUANGXI UNIV
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