Preparation method for compound enzyme system for high-efficiency degradation for lignocellulose and application thereof

A technology of lignocellulose and composite enzyme system, applied in the field of preparation of composite enzyme system, can solve the problems of unsuitability for large-scale production, low enzyme activity, poor operability, etc., and achieve broad application prospects and application potential, reducing sugar content The effect of increasing and reducing production costs

Inactive Publication Date: 2013-03-27
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ventilation effect of solid-state fermentation is not very good, the operability is poor, and it is not suitable for industrial scale-up
The carbon source used for enzyme production under liquid fermentation conditions is microcrystalline cellulose or pulp. Although the enzyme activity is high, the cost is high, the environment is seriously polluted, and it is not suitable for large-scale production; there are also cheap raw materials such as bran, straw and wheat. Straw, but the enzyme activity is low; there is also acid-treated corncob residue as raw material for fermentation, because the residue after acid treatment contains toxic by-products such as furfural, which can inhibit the growth of bacteria, so the enzyme activity is low
None of the existing production methods for cellulase can meet the requirement of producing a relatively complete enzyme system while ensuring high enzyme activity.

Method used

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  • Preparation method for compound enzyme system for high-efficiency degradation for lignocellulose and application thereof
  • Preparation method for compound enzyme system for high-efficiency degradation for lignocellulose and application thereof
  • Preparation method for compound enzyme system for high-efficiency degradation for lignocellulose and application thereof

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preparation example Construction

[0019] The invention provides a preparation method of a composite enzyme system for efficiently degrading lignocellulose, comprising the following steps:

[0020] Step 1, activating Trichoderma reesei strain or its derivative strain;

[0021] Step 2. Prepare the activated Trichoderma reesei strain or its derivative strains into a spore suspension, inoculate it in the seed medium for cultivation, and shake it on a shaking table with a temperature of 28-30°C and a rotational speed of 170-200 rpm Cultivate for 24~36h;

[0022] Step 3: Inoculate the seed culture medium inoculated with the Trichoderma reesei strain or its derivative strains into a fermentation medium with microcrystalline cellulose and corncobs as carbon sources and a mixing ratio of 1:4 for fermenter cultivation.

[0023] In the preparation method of the described compound enzyme system for efficiently degrading lignocellulose, in the step 3, the seed culture medium inoculated with the Trichoderma reesei strain o...

Embodiment 1

[0045] The activated Trichoderma reesei strain or its derivative strain on the PDA slant medium is prepared into a spore suspension, which is inoculated into a fresh seed medium according to the first percentage of inoculation amount for cultivation. Mix microcrystalline cellulose and corncob in different proportions, take 1.5g raw materials with different mixing proportions and add 50ml fermentation medium solution into a 250ml Erlenmeyer flask, sterilize at 120°C for 15min, inoculate Trichoderma reesei strain or its The seed medium solution of the derived strain was fermented at 28-30°C, 150-220 rpm for 5-7 days, and the supernatant was collected by centrifugation to measure the activity of cellulase and xylanase. The results are shown in Table 1. When microcrystalline cellulose and corn cob were mixed at a ratio of 2:8, the cellulase activity was 94.6% of that when pure microcrystalline cellulose was used as carbon source, and the xylanase activity was 94.6% of pure microcry...

Embodiment 2

[0049] According to the implementation method in Example 1, the activated Trichoderma reesei strain and its derivative strains were inoculated in the seed medium for cultivation, then inserted into 3L fermentation medium solution containing 150g / L of mixed raw materials and packed into 5l Fermentation in the fermenter, after the fermentation, the cellulase activity and xylanase activity were measured, the results are shown in figure 1 , the cellulase activity reached 9.7IU / ml after 96h, and the xylanase activity reached 3000IU / ml after 132h.

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Abstract

The invention discloses a preparation method for a compound enzyme system for high-efficiency degradation for lignocellulose and an application thereof. The preparation method comprises the following steps of: step 1, activating Trichoderma reesei strains or the derivative strains thereof; step 2, preparing spore suspension from the activated Trichoderma reesei strains or the derivative strains thereof, inoculating into a seed culture medium, and culturing for 24-36 hours in a shaking manner on a shaking table with a rotational speed of 170-200 r/min and at a temperature of 28-30 DEG C; and step 3, inoculating the seed culture medium inoculated with the Trichoderma reesei strains or the derivative strains thereof into a fermentation culture medium using microcrystalline cellulose and corncobs as a carbon source in a mixing ratio of 1:4, and performing fermentation tank culture. The compound enzyme system is used for treating corncobs, gas-detonating straw and the corn straw pre-treated by NaOH at a normal temperature. Via the method disclosed by the invention, the saccharification and hydrolysis capacities of the compound enzyme system are enhanced, the preparation cost of enzymes is reduced, and the pollution to environment is low.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a preparation method and application of a composite enzyme system for efficiently degrading lignocellulose. Background technique [0002] With the depletion of non-renewable fossil energy, more and more researches are focused on the development and utilization of new energy. Biomass energy is favored due to its abundant sources, low cost and renewable nature. As the most abundant renewable resource on earth, cellulose provides a rich source of biomass energy. According to reports, the total amount of cellulose produced by photosynthesis in the world every year can reach 10 11 Ton. However, cellulose is a linear high-molecular polymer composed of glucose molecules bound by 1,4-glycosidic bonds. Cellulose molecules are difficult to be utilized directly and must be degraded into small molecular sugars before they can be utilized. Cellulose can be degraded directly by acid, alkali ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12P19/14C12R1/885
Inventor 武改红陈树林马立娟张东远李德茂马延和
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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