Non-immetsed type liquid fermentation process of lingin catabolic enzymes from Phanerochaete chrysosporium

A technology of lignin-degrading enzymes and Phanerochaete, which is applied in the field of applied microorganisms, can solve the problems that are not conducive to the large-scale production of lignin-degrading enzymes, reduce the feasibility of fermentation, increase the activity of proteases, etc., and achieve fermentation conditions that are easy to control , Reduce fermentation cost, easy to harvest

Inactive Publication Date: 2005-01-12
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although oxygen-enriched conditions are beneficial to the synthesis of lignin-degrading enzymes of Phanerochaete chrysosporium, they may also increase the activity of proteases, resulting in rapid decomposition and instability of lignin-degrading enzymes.
Moreover, the implementation of oxygen-enriched conditions will increase the cost of fermentation and reduce the feasibility of fermentation, which is not conducive to the realization of large-scale production of lignin-degrading enzymes.
At present, there is no mature form of liquid fermentation of Phanerochaete chrysosporium lignin-degrading enzymes that can be effectively carried out under ambient air conditions.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Prepare a carbon-limited (carbon-to-nitrogen ratio 3.8:1) medium containing 5 g / L of glucose, 4.1 g / L of ammonium tartrate, 2.0 g / L of potassium dihydrogen phosphate, 0.5 g / L of magnesium sulfate, and 0.1 g of calcium chloride / L, vitamin B1 1mg / L, and appropriate amount of trace elements. A 0.5cm polyurethane foam block was used as the immobilized culture carrier. After boiling, washing and drying the carrier, put it into a 250ml Erlenmeyer flask (containing 100ml culture medium). The amount of the carrier was 3.0g / 250ml Erlenmeyer flask. The height of the liquid level was 5 times, and the medium and the carrier were sterilized at 113°C for 30 minutes. Aseptically inoculate Phanerochaete chrysosporium BKM-F-1767 spores grown on 30°C potato dextrose agar plate (200g / L potato juice, 20g / L glucose and 20g / L agar) into the culture medium, inoculum size 1×10 5 Spores / ml. The Erlenmeyer flask was placed on a rotary shaker, and the fermentation was carried out in an air en...

Embodiment 2

[0011] Prepare a nitrogen-limited (carbon-to-nitrogen ratio 152.7:1) medium containing 10 g / L of glucose, 0.2 g / L of ammonium tartrate, 2.0 g / L of potassium dihydrogen phosphate, 0.5 g / L of magnesium sulfate, and 0.1 g of calcium chloride / L, vitamin B1 1mg / L, and appropriate amount of trace elements. A 0.5cm polyurethane foam block was used as the immobilized culture carrier, and the carrier was boiled, washed and dried and then put into a 250ml Erlenmeyer flask (containing 100ml culture medium). The amount of the carrier was 2.2g / 250ml Erlenmeyer flask. The height of the liquid level was 3 times, and the medium and the carrier were sterilized together at 113° C. for 30 minutes. Aseptically inoculate Phanerochaete chrysosporium BKM-F-1767 spores grown on 30°C potato dextrose agar plate (200g / L potato juice, 20g / L glucose and 20g / L agar) into the culture medium, inoculum size 1×10 5 Spores / ml. The Erlenmeyer flask was placed on a rotary shaker, and the fermentation was carr...

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PUM

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Abstract

This invention relates to thelephoroid lignin non-immersion liquid fermentation method which inlets the thelephoroid liquid seeds in a liquid culture medium with carrier to be cultivated in oscillation under air atmosphere of 35-39deg.C. Said solid cultivation carrier is either inert or non-inert and a 0.5-2.5cm size block. Stacked height of the carrier is higher than the surface of the liquid culture medium for about 1-5times higher and the carrier is at un-immersion state. Said fermentation culture medium adopts either carbon limit liquid medium or nitrogen limit liquid culture medium PH is controlled at 4.0-6.0 the medium contains reratryl alcohol of 0-2.5mM.

Description

technical field [0001] The invention relates to a fermentation method of Phanerochaete chrysosporium lignin degrading enzyme, belonging to the field of applied microorganisms. Background technique [0002] Phanerochaete chrysosporium can produce and secrete lignin-degrading enzymes extracellularly to degrade lignin and many persistent organic pollutants, which has important application value in environmental pollution control. Phanerochaete chrysosporium lignin-degrading enzyme fermentation is mainly divided into liquid fermentation and solid-state fermentation. Compared with solid-state fermentation, liquid fermentation has good mass transfer effect, easy control of fermentation conditions, easy recovery of fermentation products and suitable for scale-up and other advantages, but the oxygen supply will be greatly affected during the fermentation process, and the enzyme yield will be limited. The expression of lignin-degrading enzymes of Phanerochaete chrysosporium is gener...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24
Inventor 文湘华喻国策熊小平钱易
Owner TSINGHUA UNIV
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