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Method for extracting cellulase from microorganism fermentation broth

A microbial fermentation liquid and microbial fermentation technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high cost, complicated operation, and short time-consuming cellulase, and achieve short time-consuming and simple operation , the effect of small energy consumption

Inactive Publication Date: 2013-01-16
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for extracting cellulase from microbial fermentation broth against the disadvantages of the general method for extracting cellulase. Cellulase, and use the vacuum freeze-drying method to prepare powdered cellulase products, so that the recovery rate of cellulase can reach 90-95%, and it has the advantages of short time, low energy consumption, simple operation, and high yield of cellulase Advantages, overcome the high cost and complicated operation of the whole lignocellulose ethanol production process in the stage of separation and extraction of cellulase, and are suitable for large-scale industrial production of cellulase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The fermented liquid submerged by Trichoderma reesei for 200 hours was first clarified through a plate and frame filter press equipped with 5 layers of gauze for primary clarification, and then through a 10 μm filter cloth for secondary clarification. .

[0040] Adjust the pH of the clarified fermentation broth to 4.0, and adjust the rotation speed to 200r / min. Weigh 3g / L tannin and fully dissolve it with pH4.0 citric acid buffer solution, then add it to the mixing tank at one time, stir it evenly and let it stand for 1h. The fermented broth after tannin precipitation was collected by press filtration with a 10 μm filter cloth plate frame, and the precipitate was collected by vacuum freeze dryer at a vacuum degree of 11 Pa and at -50°C for 30 hours to obtain a powdered cellulase product. The enzyme activity of the fermentation broth was measured by the international standard method recommended by the International Union of Theoretical and Applied Chemistry, and the yie...

Embodiment 2

[0042]The fermented liquid submerged by Trichoderma reesei for 200 hours was first clarified through a plate and frame filter press equipped with 5 layers of gauze for primary clarification, and then through a 10 μm filter cloth for secondary clarification.

[0043] Adjust the pH of the clarified fermentation broth to 4.5, and adjust the rotation speed to 150 r / min. Weigh 5g / L tannin and fully dissolve it with pH4.5 citric acid buffer solution, then add it to the stirring tank at one time, and keep stirring for 2h. The fermented broth after tannin precipitation was collected by press filtration with a 10 μm filter cloth plate frame, and the precipitate was collected by vacuum freeze dryer at a vacuum degree of 11 Pa and at -50°C for 30 hours to obtain a powdered cellulase product. The enzyme activity of the fermentation broth was measured by the international standard method recommended by the International Union of Theoretical and Applied Chemistry, and the yield of the enzym...

Embodiment 3

[0045] The fermented liquid submerged by Trichoderma reesei for 200 hours was first clarified through a plate and frame filter press equipped with 5 layers of gauze for primary clarification, and then through a 10 μm filter cloth for secondary clarification.

[0046] Adjust the pH of the clarified fermentation broth to 5.0, and adjust the rotation speed to 100 r / min. Weigh 7g / L tannin and fully dissolve it with pH5.0 citric acid buffer, then slowly and evenly add it to the stirring tank, and continue stirring for 4h. The fermented broth after tannin precipitation was collected by press filtration with a 10 μm filter cloth plate frame, and the precipitate was collected by vacuum freeze dryer at a vacuum degree of 11 Pa and at -50°C for 30 hours to obtain a powdered cellulase product. The enzyme activity of the fermentation broth was measured by the international standard method recommended by the International Union of Theoretical and Applied Chemistry, and the yield of the enz...

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Abstract

The invention discloses a method for extracting cellulase from microorganism fermentation broth, which comprises the following steps of: fermentation broth clarification: carrying out filter pressing on fermentation broth which is fermented by microorganism for generating cellulase for about 140-200 hours to obtain fermentation clarifying solution, and transferring the fermentation clarifying solution to a stainless steel agitation tank; tannin precipitation: firstly regulating revolving speed to stir the clarified fermentation broth, meanwhile, regulating pH (potential of hydrogen) to be between 4.0 and 6.0, then fully dissolving a certain quantity of tannin with citrate buffer solution, and slowly and evenly adding the mixture into the agitation tank to be continuously stirred for a certain period of time; and precipitation collection and preparation: carrying out filter pressing, collection and precipitation on the fermentation broth precipitated by the tannin via a plate-and-frame filter press, and carrying out freeze-drying by a vacuum freezer dryer to obtain a powder cellulase product. When being used for extracting the cellulase, the method disclosed by the invention has the advantages of short use time, small energy consumption and simplicity in operation, and the yield of cellulase protein can be 90-95%.

Description

technical field [0001] The invention relates to a method for separating and extracting cellulase in industry, in particular to a method for extracting cellulase from microbial fermentation liquid. Background technique [0002] Cellulose resources are the cheapest and most abundant renewable resources in the world. my country is a large agricultural country, and the reserves of lignocellulose such as wheat straw are even more abundant. In many rural areas, wheat stalks are even burned, which not only causes serious environmental pollution but also causes a lot of waste of lignocellulosic raw materials. Therefore, how to rationally utilize this renewable resource has become an urgent problem, and workers from various countries have done a lot of research on solving this problem. [0003] At present, the production of fuel ethanol from lignocellulose by cellulase is one of the most effective methods for large-scale utilization of lignocellulose, and it is also a hot spot in n...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12R1/885
Inventor 宗志友马立娟赫荣琳李晨贾文娣陈树林
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI