Bacterial strain for producing laccase, method for producing laccase by bacterial strain, produced laccase and application of laccase

A technology of laccase and bacterial strains, which is applied in the field of microbial enzymology, can solve the problems of weak tolerance, hindrance to large-scale commercial application, low activity and yield of laccase, and achieve easy operation, high tolerance and high activity. Effect

Inactive Publication Date: 2015-05-13
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology describes an improved process that produces Lacases from Bacillus brevis or other microorganisms without causing damage during culturing. These processes have higher activities than existing methods but they still require harsher environments like pH 7.5-7.5. They produce less harmful substances compared to traditional chemical treatments. Additionally, these new techniques could potentially improve various applications related to lignin products including use them in paper manufacture. Overall, this innovative technique offers technical benefits over current technologies while reducing costs associated therewith.

Problems solved by technology

Technologies aim at developing new methods or materials to treat contaminated waters containing chemical substances like pesticides, heavy metals, and pharmaceutically active agents called synthetic hydrocresispersive ligands (SL). These technical problem addressed include finding effective ways to prevent damage to aquifera communities, reducing negative impact on wildlife, protecting sensitive areas around us, promoting welfare activities, creating valuable products without causing secondary pollutions, and providing practicality guidelines for applying SL in wider ranges of technologically demandable markets.

Method used

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  • Bacterial strain for producing laccase, method for producing laccase by bacterial strain, produced laccase and application of laccase
  • Bacterial strain for producing laccase, method for producing laccase by bacterial strain, produced laccase and application of laccase
  • Bacterial strain for producing laccase, method for producing laccase by bacterial strain, produced laccase and application of laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 culture medium

[0048] 1. Separation and purification medium

[0049] Glucose 20.0g / L, malt extract powder 20.0g / L, agar 25.0g / L, KH 2 PO 4 3.0g / L, penicillin 0.2g / L, streptomycin 0.2g / L, deionized water 1000mL, pH natural, 1×10 5 Pa autoclave for 30 minutes, add penicillin and streptomycin when the temperature is lowered to ≤40°C, shake well, distribute in plates, and store at 4°C after cooling for later use.

[0050] 2. Preserve slant culture medium

[0051] Glucose 20.0g / L, malt extract powder 20.0g / L, agar 25.0g / L, KH 2 PO 4 3.0g / L, 1000mL deionized water, natural pH; 1×10 5 After Pa autoclaving for 30 minutes, place the test tube rack at an angle to make the culture medium slope, and store it at 4°C after cooling for later use.

[0052] 3. Activation medium (solid plate medium)

[0053] Glucose 20.0g / L, yeast extract powder 5.0g / L, agar powder 20.0g / L, KH 2 PO 4 1.0g / L, MgSO 4 ·7H 2 O0.5g / L, ZnSO 4 ·7H 2 O 50mg / L, vitamin B110mg / L, deionize...

Embodiment 2

[0056] Example 2 Isolation, purification and identification of Trametes pubescens BJFC-C1108

[0057] 1. Isolation and purification of Trametes pubescens BJFC-C1108

[0058] The bacterial strain of the present invention is isolated from fallen logs of Castanopsis chinensis in Chebaling Nature Reserve of Guangdong Province.

[0059] The white rot fungus Trametes tomentosa was collected from the fallen logs of Castanopsis chinensis in Chebaling Nature Reserve, Guangdong Province. The fresh fruiting body was carefully peeled off from the substrate with a collection knife, and impurities irrelevant to the fruiting body were removed as much as possible. workbench ( ), cut the fungus flesh near the base of the fruiting body (the size is about 3.5cm×3.5cm×3.5cm, and the vegetative hyphae at the base of the fruiting body grow the most vigorously) and inoculate it on the separation and purification plate medium, at 28±3 After static culture at constant temperature at ℃ for 7-8 days,...

Embodiment 3

[0068] Example 3 Preparation and purification of laccase Tplac by fermenting Trametes pubescens BJFC-C1108

[0069] 1. Preparation of Trametes villosa bacteria cake

[0070] Inoculate the purified Trametes pubescens BJFC-C1108 strain on the activated solid medium, culture it statically at 28±3°C for 7-8 days, and then pass through a hole puncher with a diameter of 1 cm to make the villous plug hole Bacteria Cake.

[0071] 2. Preparation of fermented seed suspension

[0072] Inoculate the bacteria cake of Trametes versicolor into a 250mL Erlenmeyer flask containing 100mL of liquid medium, and culture it with shaking at 28±3°C and 150rpm. After 6 days, use an in-cut homogenizer at 5000 rpm for 30 seconds to homogenate, and oscillate fully to obtain a suspension of fermented seeds of Trametes villiformis.

[0073] 3. Fermentation culture

[0074] The fermented seed suspension of Trametes villiformis was added to a 250mL Erlenmeyer flask containing 100mL liquid fermentation me...

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Abstract

The invention discloses a bacterial strain for producing laccase, a method for producing laccase by the bacterial strain, the produced laccase and application of the laccase, and relates to the field of microbial enzyme. The bacterial strain for producing laccase is trametes pubescens (Trametes pubescens BJFC-C1108) with a microbial preservation number of CGMCC No. 10308. By utilizing the trametes pubescens (Trametes pubescens BJFC-C1108), the yield of the produced laccase is relatively high; the enzyme activity can be up to 28.687 U/mL; the prepared laccase has relatively high activity, stability and tolerance under the alkali condition (pH 7.0-10.0), has relatively high tolerance for metal ions, can be used for shortening the time required by complete decoloring of azo dye Congo red to a certain extent, has a potential application value, can be applied to more industrial fields and has favorable development and application prospects.

Description

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Claims

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Application Information

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Owner BEIJING FORESTRY UNIVERSITY
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