Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A kind of thermostable Trametes trichos laccase ttlac13 and its application

A thermostability, technology of Trametes trichotis, applied in the field of enzyme engineering and genetic engineering, can solve the problem of low thermal stability of laccase strain Trametes trichosum, achieve good industrial application prospects, good economic benefits and social effects, improve efficiency effect

Active Publication Date: 2018-12-14
KUNMING UNIV OF SCI & TECH
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the existing research, the thermal stability of the laccase from the purified Trametes trichotillotype strain is not high

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of thermostable Trametes trichos laccase ttlac13 and its application
  • A kind of thermostable Trametes trichos laccase ttlac13 and its application
  • A kind of thermostable Trametes trichos laccase ttlac13 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: Laccase produced by Trametes trichotillosum and the purification of laccase TtLac13

[0034] The laccase strains used were screened by our laboratory and identified as T. trogii For typical strains, put this strain in a medium containing glucose 20 g / L, yeast extract 5 g / L, peptone 5 g / L, MgSO 4 .7H 2 O 1 g / L, plus 15 g / L agar on a solid medium plate or a large test tube for bacterial strain activation. After cultivating for 3 days, break the activated bacterial strain with sterilized glass beads, and then take the broken The final bacterial solution was placed in a solution containing 28 g / L of glucose, 10 g / L of yeast extract, 10 g / L of peptone, FeSO 4 0.4 mM, CuSO4 1.65 mM, Tween-80 6 g / L, PEG 4000 45 mg / L in the optimized enzyme-producing liquid medium, make three groups and place them in a shaker at 28°C at 180rpm.

[0035]Collect the above cultured culture medium, centrifuge at 5000 rpm, 4 °C for 5 min to collect the supernatant, which is the cru...

Embodiment 2

[0036] Example 2: Determination of protein concentration

[0037] The protein concentration at each stage of the laccase purification process was determined by the Coomassie Brilliant Blue method. Take 0.1 mL of properly diluted purified laccase and add it to 5 mL of Coomassie Brilliant Blue G-250 reagent, that is, add 95% ethanol to a mortar with 100 mg of Coomassie Brilliant Blue G-250 and grind to dissolve it, and take the upper layer The liquid was placed in a beaker, and 10 mL of 95% ethanol was added and ground in a mortar, repeated 3 times to dissolve the Coomassie brilliant blue, and the mortar was washed with 20 mL of ethanol several times. Add 100 mL of 85% concentrated phosphoric acid to the beaker in portions, and then dilute to 1 L with deionized water. After reacting for 2-5 minutes, measure the light absorption value of the reaction solution at 595 nm on a spectrophotometer. To draw a standard curve, add 1 mg / L standard protein solution containing 100 mg / L bov...

Embodiment 3

[0038] Embodiment 3: enzyme activity assay method

[0039] Using ABTS as the substrate, the increase in the absorbance value of the reaction solution at 420 nm was measured within 3 min. First prepare 100 mM phosphate-citrate buffer, pH 4.0. And use this buffer to prepare 2 mM ABTS solution. Use 1.5 mL reaction system: 1.1 mL phosphate-citrate buffer (100 mM, pH 4.0) containing 2 mM ABTS and 0.4 mL laccase pure enzyme diluted with phosphate-citrate buffer (100 mM, pH 4.0) liquid. Record the change of the absorbance value of the reaction solution at 420 nm within 3 min of reaction. Define the amount of enzyme needed to oxidize 1 μmol ABTS in one minute as 1 enzyme activity unit (U). The pure enzyme solution of laccase after heat inactivation was used as the control, and all experiments were repeated in triplicate.

[0040] Laccase activity calculation formula U(U / L) = N×V 1 ×(A 3 -A 0 ) / (ε×L) / V 2 / t

[0041] N is the dilution factor, V 1 is the total reaction volume,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the field of applied microbiology and the technical field of bioengineering, and particularly discloses trametes trogii laccase isozyme TtLac13. The amino acid sequence of the trametes trogii laccase isozyme TtLac13 is shown in SEQ ID NO:1. When the trametes trogii laccase isozyme TtLac13 is applied to biodegradation of triphenylmethane colored and toxic substances such as peacock green and crystal violet, time is short, decoloration efficiency is high, good industrial application prospects are achieved, and good economic benefits and good social benefits can be generated.

Description

technical field [0001] The present invention relates to a kind of Trametes trichotis ( Trametes trogii ) laccase isoenzyme TtLac13 and its application, the laccase has high thermal stability, high tolerance to acid, alkali, organic solvent and heavy metal ions, and has great effect on various colored and toxic substances such as crystal violet and malachite green Strong degradability, belongs to the field of enzyme engineering and genetic engineering. Background technique [0002] Laccase (EC1.10.3.2, Laccase, lac) is a copper-containing polyphenol oxidase belonging to the blue multicopper oxidase family, which can convert various phenolic and nonphenolic It is an "eco-friendly" enzyme that oxidizes substrates to generate water. Laccase has a wide range of substrates and high catalytic efficiency. It is widely used in the fields of biopulping, food industry, wood processing, biosynthesis, bioenergy, biodetection and bioremediation, such as the treatment of paper lignin, th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02
CPCC12N9/0061C12Y110/03002
Inventor 严金平伍圆圆高升伊日布斯杨恩陈带娣牛杰振
Owner KUNMING UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com