Method for replenishing nitrogen source culture fungus in batch to produce cellulase

A cellulase and production method technology, applied in the field of enzymatic fermentation, can solve the problems of high mass transfer, increased osmotic pressure, unfavorable bacterial growth in the lag phase, etc., and achieves the effects of reducing production cost, facilitating mycelial absorption, and promoting mass synthesis

Inactive Publication Date: 2014-12-03
熊鹏
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, this method adds the nutrients needed in the entire fermentation process at one time in the early stage, which will easily lead to the excessive concentration of the initial medium, resulting in difficulties in mass transfer and heat transfer, and increasing the energy consumption of fermentation; the high concentration of nutrients in the early stage of fermentation will cause the entire The increase of the osmotic pressure of the fermentation medium is not conducive to the growth of the strains in the lag phase; the high concentration of nutrients will also cause a large amount of nutrients in the early stage to be used for the nutrients needed for the growth of the synthetic strains, resulting in a large number of bacterial strains, so that the late fermentation of synthetic fibers Nutrient deficiency inhibits enzyme production

Method used

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  • Method for replenishing nitrogen source culture fungus in batch to produce cellulase
  • Method for replenishing nitrogen source culture fungus in batch to produce cellulase
  • Method for replenishing nitrogen source culture fungus in batch to produce cellulase

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1: take Trichoderma reesei Rutc-30 as fermentation strain

[0027] In a 50L fermenter, pretreated rice straw was used as the only carbon source for enzyme production and a medium was prepared with an enzyme inducer. The medium components were: pretreated straw 30g / L, corn steep liquor (CSL) 3g / L , (NH 4 ) 2 SO 4 1.4 g / L, KH 2 PO 4 2 g / L, MgSO 4 ·7H 2 O 0.3g / L, FeSO 4 ·7H 2 O 5mg / L, ZnSO 4 ·7H 2 O 1.4mg / L, MnSO 4 ·7H 2 O 1.6mg / L, CoCl 2 ·6H 2 O 2mg / L, CaCl 2 0.01g / L; Sterilize at 121°C for 20 minutes, cool down to 30°C, inoculate 2L of cultured Rutc-30 seeds with 5% inoculum in the sterilized fermenter, and ferment at 30°C for 7 days to produce enzymes. Fermentation ends , the highest measured enzyme activity is 2.12u / ml, the growth and enzyme production curve is as follows figure 1 shown.

Embodiment 2

[0028] Embodiment 2: Taking Trichoderma reesei Rutc-30 as fermentation strain

[0029] In a 50L fermenter, pretreated rice straw was used as the only carbon source for enzyme production and a medium was prepared with an enzyme inducer. The medium components were: pretreated straw 30g / L, corn steep liquor (CSL) 1g / L , (NH 4 ) 2 SO 4 1.4 g / L, KH 2 PO 4 2 g / L, MgSO 4 ·7H 2 O 0.3g / L, FeSO 4 ·7H 2 O 5mg / L, ZnSO 4 ·7H 2 O 1.4mg / L, MnSO 4 ·7H 2 O 1.6mg / L, CoCl 2 ·6H 2 O 2mg / L, CaCl 2 0.01g / L; sterilize at 121°C for 20min, cool down to 30°C, inoculate 1L of the cultivated Rutc-30 seeds with 2.5% inoculum in the sterilized fermenter, and ferment on the 3rd and 5th day respectively Add 100ml of 100g / L corn steep liquor solution, and ferment at 30°C for 7 days to produce enzyme; the highest enzyme activity is 3.21u / ml, and the growth and enzyme production curve is as follows: figure 2 shown.

Embodiment 3

[0030] Embodiment 3: take Trichoderma reesei Rutc-30 as fermentation strain

[0031] In a 50L fermenter, pretreated rice straw was used as the only carbon source for enzyme production and a medium was prepared with an enzyme inducer. The medium components were: pretreated straw 30g / L, corn steep liquor (CSL) 3g / L , (NH 4 ) 2 SO 4 0.5 g / L, KH 2 PO 4 2 g / L, MgSO 4 ·7H 2 O 0.3g / L, FeSO 4 ·7H 2 O 5mg / L, ZnSO 4 ·7H 2 O 1.4mg / L, MnSO 4 ·7H 2 O 1.6mg / L, CoCl 2 ·6H 2 O 2mg / L, CaCl 20.01g / L; sterilize at 121°C for 20min, cool down to 28°C, inoculate 400ml of cultured Rutc-30 seeds with 1% inoculum in the sterilized fermenter; Add 100g / L (NH 4 ) 2 SO 4 100ml, fermented at 28°C for 7 days to produce enzymes; the enzyme activity was measured up to 2.85u / ml, and the growth and enzyme production curves were as follows: image 3 shown.

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Abstract

The invention discloses a method for replenishing nitrogen source culture fungus in batch to produce cellulose, which comprises the following steps: sterilizing culture medium in a fermentation tank in 60-80 percent loaded liquid at 121 DEG C for 20 min, and cooling to culture temperature of 28-30 DEG C; inoculating fungus seed solution in an inoculum size of 1-5 percent loaded liquid to ferment in a fermentation tank; and replenishing inorganic nitrogen source and organic nitrogen source of which quality is 0.25-0.5 times of correspondence quality of nitrogen source in initial culture medium at second to third day and fourth to fifth day in fermenting and culturing respectively. The invention has the advantages that osmotic pressure of fermentation liquor is reduced through adding little amount of N source in early days, nutrition of initial culture medium can be reduced, excess reproduction of hypha can be inhibited and abundant nutrition for hypha growth can be avoided; through replenishing nitrogen source in batch at the later period of fermentation, nutrition supply desired to compound cellulose can be ensured while nutrition for hypha growth is limited; the utilization ratio of nutrition can be increased; and higher fermentation cellulose activity can be obtained.

Description

technical field [0001] The invention belongs to the field of enzyme fermentation, and in particular relates to a method for fermenting and producing cellulase by supplementing nitrogen sources in batches during the fermentation process. technical background [0002] The ability of cellulase to hydrolyze lignocellulose into sugar makes it have a wide application prospect. By hydrolyzing lignocellulose such as straw, fermenting it into sugar, and preparing fuel ethanol or other chemical products through fermentation and other methods, This is also one of the key elements in the preparation of energy and chemical products from lignocellulose. The cost and efficiency of cellulase preparation are also one of the decisive factors for whether biomass-based energy and chemical products can gradually replace petroleum energy and chemical products. [0003] The high-activity cellulase fermentation technology can greatly improve the fermentation efficiency and save the cost of enzyme p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12R1/885C12R1/66
Inventor 熊鹏宇文伟刚贺建龙
Owner 熊鹏
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