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Cellulose-degradation fungus and preparation of inoculum and application thereof

The technology of spore inoculum and strain is applied in the field of microorganisms, which can solve the problem that microbial degradation is difficult to achieve the expected effect, and achieve the effects of excellent fermentation properties, low cost and simple fermentation process.

Active Publication Date: 2015-04-01
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since silkworm excrement contains a large amount of cellulose that is difficult to degrade, it is difficult to achieve the desired effect simply by relying on microbial degradation in silkworm excrement.

Method used

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  • Cellulose-degradation fungus and preparation of inoculum and application thereof
  • Cellulose-degradation fungus and preparation of inoculum and application thereof
  • Cellulose-degradation fungus and preparation of inoculum and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Enrichment and Isolation of Cellulose Degrading Bacteria

[0043] Take 5g of the humus soil sample collected from the Taishan forestland and add it to 45mL enrichment medium, shake it at 30°C and 180r / min for 3 days, then transfer 5mL culture solution to another triangle filled with fresh enrichment medium Continue to culture in the bottle for 3 days.

[0044] Take the enriched culture solution and dilute it with sterile water to 10 -4 、10 -5 、10 -6 、10 -7 4 concentration gradients. Use a pipette to draw 100 μL of each of the four concentrations of soil dilutions and spread them on the CMC medium plate. Repeat the coating test for each concentration three times, culture at a constant temperature of 28°C for 2 days, and pick a single colony from the medium on the CMC Separation and purification by streaking on the culture medium plate.

[0045] Transfer the strains purified on the CMC medium plate to the cellulose Congo red medium plate. Each strain was replicated ...

Embodiment 2

[0047] 1. Degradation test of filter paper

[0048] Take six 250mL Erlenmeyer flasks, numbered 1-6 respectively; among them, put 1cm×6cm filter paper strips into the 1-3 Erlenmeyer flasks, add 50mL of Hutchinson’s medium to each, and inoculate 1mL The seed solution of cellulose-degrading bacteria C2 was used as the test group; filter paper strips of 1 cm × 6 cm were put into No. As a control group; all six Erlenmeyer flasks were placed in a constant temperature shaker at 30°C, shaken at 200r / min for 5 days, and the filter paper strips in No. 1 to No. 3 Erlenmeyer flasks were visually inspected to form a paste.

[0049] 2. Determination of filter paper cellulase activity

[0050] The present invention uses 3,5-dinitrosalicylic acid (DNS) to react with reducing sugars under alkaline conditions to generate colored compounds, and conduct colorimetric measurement with a spectrophotometer to determine the amount of low-molecular sugars. cellulase activity. The reddish-brown amino...

Embodiment 3

[0065] Identification of Cellulose Degrading Bacteria C2

[0066] 1. Colony characteristics and morphological characteristics of bacteria

[0067] (1) Colony characteristics

[0068] The strain grows rapidly on the PDA plate, the mycelium is dense, the colony is flat, and forms a shell. The mycelium is white at the beginning, and as the cultivation time increases, the mycelium gradually becomes dark green, and the back of the medium is light yellow; The growth on the CMC plate is slow, the mycelium is sparse, the mycelium is also short, the mycelium is flat, and the mycelium is white at the beginning. As the cultivation time increases, the mycelium gradually turns blue-green (see figure 1 ).

[0069] (2) Morphological characteristics of bacteria

[0070] The vegetative mycelium has a septum, the conidiophores grow vertically from the mycelium, and the top grows broom-like interbranches, 7-9 small stalks, (11.5-14.8) μm × (3.0-3.4) μm, conidia Oval, smooth, size (4.8~6.0) μ...

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Abstract

The invention provides a cellulose-degradation fungus, preparation of an inoculum and an application thereof. According to the invention, a strain is obtained from Mount Taishan woodland humus soil by artificial enrichment and screening; the strain is identified as penicillium oxalicum, which is named as penicillium oxalicum C2; the strain has good cellulose degradation capability, and high cellulose-production activity; after shake flask fermentation for 3 days, the filter paper cellulose activity is up to 132.26 U / mL; When used in silkworm excrement compost, the conidium inoculums prepared from the penicillium oxalicum C2 can increase the compost temperature, prolong the high temperature stage, facilitate water content reduction, promote organic matter decomposition and the increase of total nitrogen relative content, decrease the C / N ratio, raise the seed germination index, accelerate the composting process, improve decomposition efficiency and fertilizer quality; the preparation method of the inoculums is simple in process, low in cost, and suitable for industrial production.

Description

technical field [0001] The invention relates to the preparation and application of a cellulose-degrading fungus and its bacterial agent, belonging to the technical field of microorganisms. Background technique [0002] Cellulose is the most abundant renewable energy substance in nature, and it is widely found in crop straw, livestock and poultry manure, municipal solid waste and paper mill wastewater. According to reports, the annual production of cellulose through photosynthesis in the world is as high as 4.0×10 10 tons, of which the output of crop straw is about 2.2 billion tons. As a large agricultural country, my country is rich in crop straw resources, which can reach more than 650 million tons per year, accounting for 20% to 30% of the world's total straw output. However, due to the complex structure of cellulose molecules, the natural degradation process is extremely slow. At present, the utilization efficiency of cellulose resources in my country is very low. Except...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N3/00C05F17/00C12R1/80
CPCY02W30/40
Inventor 刘训理刘鹏姜红霞周保强鲍正宗隋君康赵旭王晓辉
Owner SHANDONG AGRICULTURAL UNIVERSITY
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