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Wood-rotting basidiomycetes for production of ligninolytic enzymes

A technology for lignin-degrading enzymes and basidiomycetes, which is applied in the direction of using microorganisms, enzymes, enzymes, etc., can solve the problems of unsatisfactory yield or production level of lignin-degrading enzymes, and toxic inducers.

Inactive Publication Date: 2008-06-25
MYCOENZYME
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the lignin-degrading enzyme yields or levels of production in these and other schemes are still not satisfactory for large-scale industrial production to be profitable.
They use very expensive equipment and media components; some inducers used are toxic

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Cerrena unicolor was used as the production strain. First, malt extract agar was inoculated, and then cultured at 27°C for about 10-12 days. Remove 1-2cm from the agar surface 2 The amount of growing mycelium was inoculated in 100ml of nutrient medium in a 500ml Erlenmeyer flask (cultivation time at 27°C was about 5 days). The preculture thus obtained was then homogenized twice for 30 seconds each in a Waring laboratory mixer. Add 50ml of preculture mycelial homogenate per liter of medium in the shake flask, ie add 25ml to a 2L shake flask containing 0.5L of medium containing ethanol production waste. The culture temperature was 27°C; the shaking frequency was 140 rpm. The culture period is 7-10 days. Laccase and manganese peroxidase yields were approximately 50000-150000 IU / L and 500-800 IU / L, respectively.

[0034] The medium composition is as follows:

[0035] a) Pre-medium (g / L):

[0036] Glucose-10.0

[0037] Peptone-0.2

[0038] Yeast Extract - 0.3

[00...

Embodiment 2

[0050] (Repeat the conditions of Example 1; 0.5L main nutrient medium contains 60g / L crushed orange peel).

[0051] Cerrena unicolor was used as the strain. First, malt extract agar was inoculated, and then cultured at 27°C for about 10-12 days. Remove 1-2cm from the agar surface 2 The amount of growing mycelium was inoculated in 100ml of nutrient medium in a 500ml Erlenmeyer flask (cultivation time at 27°C was about 5 days). The preculture thus obtained was then homogenized twice for 30 seconds each in a Waring laboratory mixer. Add 50 ml of preculture mycelial homogenate per liter of medium in the shaker flask, ie add 25 ml to a 2L shaker flask containing 0.5L medium containing crushed orange peel. The culture temperature was 27°C; the shaking frequency was 140 rpm. The culture period is 6-9 days. Laccase and manganese peroxidase yields were approximately 15000-25000 IU / L and 4000-7000 IU / L, respectively.

[0052] The medium composition is as follows:

[0053] a) Pre-...

Embodiment 3

[0068] Trametes versicolor was used as the strain. First, malt extract agar was inoculated, and then cultured at 27°C for about 10-12 days. Remove 1-2cm from the agar surface 2 The amount of growing mycelium was inoculated in 100ml of nutrient medium in a 500ml Erlenmeyer flask (cultivation time at 27°C was about 5 days). The preculture thus obtained was then homogenized twice for 30 seconds each in a Waring laboratory mixer. Add 50 ml of preculture mycelial homogenate per liter of medium in the shaker flask, ie add 25 ml to a 2L shaker flask containing 0.5L medium containing crushed orange peel. The culture temperature was 27°C; the shaking frequency was 140 rpm. The culture period is 4-5 days. Laccase and manganese peroxidase yields were approximately 15000-20000 IU / L and 200-400 IU / L, respectively.

[0069] The medium composition is as follows:

[0070] a) Pre-medium (g / L):

[0071] Glucose-10.0

[0072] Peptone-0.2

[0073] Yeast Extract - 0.3

[0074] K H 2 PO ...

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Abstract

The invention relates to the production of ligninolytic enzymes, laccase and manganese peroxidase, from certain white-rot basidiomycetes fungi, using highly efficient fermentation techniques. The aim of this invention is to create a novel economically and time-effective overall procedure comprising use of specific mushroom strains, fermentation process and the isolation-purification techniques, for producing the aforesaid enzymes. In particular, a submerged fermentation of the specific strains on a variety of lignocellulosic substrates from organic wastes like waste of ethanol production from wheat grain, mandarin peels and bran is developed. Culturing conditions can be selected to modify the laccase / manganese peroxidase ratio in favour of the production of either laccase or manganese peroxidase.

Description

field of invention [0001] The present invention relates to the production of lignin degrading enzymes such as laccase, manganese peroxidase and lignin peroxidase from white rot fungi. Background of the invention [0002] White-rot fungi are characterized by a unique ability to degrade lignin, a recalcitrant woody polymer. The main enzymes associated with the lignin-degrading ability of white-rot fungi are lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (Lac), although the fungi mentioned above do not share the same set of enzymes. Recently, these fungi have been extensively studied with the aim of isolating new microorganisms that increase the secretion of lignin-degrading enzymes and enzymes with properties important for industrial applications: hazardous xenochemical Bioremediation of industrial wastewater streams contaminated by xenobiotics; biobleaching and biopulping; textile and dyeing industries; biotransformation of pharmaceuticals and other intermed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P1/02C12P21/00
CPCC12N1/22C12N9/0065C12N1/14C12N9/0061
Inventor V·埃利萨施维尔M·雷布恩
Owner MYCOENZYME
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