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Method for simultaneously increasing the yield and improving the activity of phellinus igniarius liquid fermented polysaccharide and laccase

A technology of liquid fermentation and Phellinus linteus is applied in the field of fermentation engineering to achieve the effects of increasing yield and biological activity, high added value of products and improving performance

Active Publication Date: 2014-12-10
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technology only targets single products such as biomass and flavonoids, and does not involve the simultaneous addition of specific compound exogenous stimuli at different fermentation stages according to the characteristics of the synthetic metabolic pathways of medicinal fungal polysaccharides and laccase-specific metabolites. Promoting the production and biological activity of polysaccharides and extracellular laccase in Phellinus Phellinus

Method used

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  • Method for simultaneously increasing the yield and improving the activity of phellinus igniarius liquid fermented polysaccharide and laccase
  • Method for simultaneously increasing the yield and improving the activity of phellinus igniarius liquid fermented polysaccharide and laccase
  • Method for simultaneously increasing the yield and improving the activity of phellinus igniarius liquid fermented polysaccharide and laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] 1. Fermentation culture

[0056] (1) Preparation of exogenous stimuli:

[0057] ①Preparation method of the enzymatic hydrolyzate of Rhododendron japonica leaves: After cleaning and vacuum freeze-drying the picked Rhododendron japonica leaves, weigh 100g and add 300mL of water, grind it into a slurry, sterilize at 95°C for 5min, and inactivate various enzymes; adjust the pH of the slurry to 4. 0, add a compound enzyme consisting of 1g cellulase and 1g hemicellulase, carry out enzymolysis reaction at 45°C for 1.0 hour, and inactivate the enzyme at 95°C for 6 minutes; then adjust the pH of the slurry to 6.0, and add 1.75g ​​of papaya The compound enzyme composed of protease and 1.75g ​​neutral protease was subjected to the enzymolysis reaction at 55°C for 1.5h, and the enzyme was inactivated at 95°C for 5 minutes to obtain the enzymolysis solution; The ultrafiltration membrane is ultrafiltered, and the retentate is vacuum-concentrated and then freeze-dried to obtain the e...

Embodiment 2

[0089] 1. Fermentation culture

[0090] (1) Preparation of exogenous stimuli:

[0091] ①Preparation method of enzymatic hydrolyzate of Rhododendron japonica leaves: After cleaning and vacuum freeze-drying the picked Rhododendron japonica leaves, weigh 100g and add 600mL of water, grind into a slurry, sterilize at 90°C for 8min to inactivate various enzymes; adjust the pH of the slurry to 5. 5. Add a compound enzyme composed of 1.1g cellulase and 2.2g hemicellulase, carry out enzymatic hydrolysis reaction at 40°C for 1.5 hours, and inactivate the enzyme at 90°C for 8 minutes; then adjust the pH of the slurry to 5.0, and add 0.5 Composite enzyme consisting of 1 g of papain and 1.5 g of neutral protease was subjected to enzymolysis reaction at 50°C for 1 hour, and the enzyme was inactivated at 90°C for 8 minutes to obtain the enzymolysis liquid; Ultrafiltration with a 3kDa ultrafiltration membrane, the retentate was concentrated in vacuum and then freeze-dried to obtain the enzy...

Embodiment 3

[0103] 1. Fermentation culture

[0104] (1) Preparation of exogenous stimuli:

[0105] ①Preparation method of enzymatic hydrolyzate of Rhododendron japonica leaves: After cleaning and vacuum freeze-drying the picked Rhododendron japonica leaves, weigh 100g and add 400mL of water, grind into a slurry, sterilize at 92°C for 6 minutes to inactivate various enzymes; adjust the pH of the slurry to 4. 5. Add a compound enzyme composed of 2g cellulase and 0.5g hemicellulase, carry out enzymatic hydrolysis reaction at 50°C for 0.5 hours, and inactivate the enzyme at 93°C for 5 minutes; then adjust the pH of the slurry to 7.0, and add 1.12g of The complex enzyme composed of papain and 0.56g neutral protease was subjected to enzymolysis reaction at 60°C for 1.2h, and the enzyme was inactivated at 92°C for 6 minutes to obtain the enzymolysis solution; Ultrafiltration with a 2kDa ultrafiltration membrane, the retentate was concentrated in vacuum and then freeze-dried to obtain the enzyma...

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Abstract

The invention discloses a method for simultaneously increasing the yield and improving the activity of phellinus igniarius liquid fermented polysaccharide and laccase. The method comprises the following steps: preparing rhododendron ovatum leaf zymolyte by using a biological compound enzyme method; inoculating the rhododendron ovatum leaf zymolyte with a culture medium with cirmamonun camphora through pleurotus tuber-regium; culturing so as to obtain a cirmamonun camphora degradation product; performing the first stage fermentation culture, namely, performing fermentation culture in the phellinus igniarius liquid and adding the rhododendron ovatum leaf zymolyte and glutamine on the first day to the fifth day; performing the second stage fermentation culture, namely, continuously culturing, and adding the rhododendron ovatum leaf zymolyte and alpha-pinene on the second day to the sixth day; and subsequently continuously culturing for 1-5 days, thereby obtaining fermentation broth of the phellinus igniarius liquid. According to the characteristics of a synthesis metabolic pathway of a medical fungus phellinus igniarius liquid fermented polysaccharide and a specific laccase metabolism product, by adding composite exogenous stimuli of different ratios in different fermentation procedures, the synergistic effects of the exogenous stimuli are brought into play, so that the yield and the bioactivity of intercellular polysaccharide and extracellular laccase of phellinus igniarius are greatly improved simultaneously, high product additional values are achieved, and the method has great industrialization prospect.

Description

technical field [0001] The invention belongs to the field of fermentation engineering, and in particular relates to a method for simultaneously increasing the output and activity of polysaccharides and laccases fermented from Phellinus phellifolia. Background technique [0002] Phellinus igniarius belongs to Basidiomycotina, Phellinus class, Rustiaceae, Pinifera. Phellinus extract has significant effects in the clinical application of inhibiting cancer cell metastasis and preventing cancer recurrence after surgery. It is currently a medicinal fungus with the highest efficiency among internationally recognized biological cancer treatment agents. [0003] Due to the particularity and complexity of the physiological state and the constraints of the external environment, Phellinus seldom forms fruiting bodies in nature, especially it takes many years to form usable fruiting bodies. Moreover, artificial cultivation is extremely difficult, the cultivation conditions are harsh, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12N9/02C12R1/645
Inventor 程俊文贺亮胡传久魏海龙付立忠李海波邹景泉
Owner ZHEJIANG FORESTRY ACAD
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