A method for preparing laccase from Candida tropicalis dk2 strain

A technology of Candida tropicalis and strains, applied in the field of biological textiles, can solve the problems of high energy consumption of chemical degumming process, unrecyclable waste water, large equipment loss, etc., and achieves good application prospects, high degumming efficiency, and good quality effects.

Inactive Publication Date: 2018-08-03
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The scouring method commonly used in the hemp industry is mainly the chemical method, that is, the raw hemp or roving is treated with strong acid and alkali. The chemical degumming process consumes a lot of energy and equipment loss, and the discharged wastewater cannot be recycled, causing serious pollution problems.

Method used

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  • A method for preparing laccase from Candida tropicalis dk2 strain
  • A method for preparing laccase from Candida tropicalis dk2 strain
  • A method for preparing laccase from Candida tropicalis dk2 strain

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Effect test

Embodiment 1

[0026] The method for preparing laccase from Candida tropicalis DK2 strain:

[0027] (1) Mycelia suspension: Candida tropicalis DK2 strain was transferred from PDA slant medium into a 300mL Erlenmeyer flask containing 50mL of basal liquid medium, and cultured in a shaker at 180rpm at a constant temperature of 30°C for 3 days, then fermented The liquid was filtered through sterile gauze, the mycelium was washed with sterile water and dispersed in a Waring high-speed crushing cup for 15 seconds, collected in sterile water (equal to the volume of the fermentation broth) to make a suspension of mycelia, and collected at 40 Store at ℃ for later use.

[0028] Basic liquid medium: 200g potato, 20g glucose, 1000mL distilled water, natural pH.

[0029] (2) Enzyme production in shake flasks: add 1% (w / v) natural substrate) and different concentrations of CuSO in the base liquid medium 4 ·5H 2 O (0-4mM) is the enzyme production medium. Put 80mL of enzyme-producing medium into a 300mL...

Embodiment 2

[0033] The process of degumming flax, jute, kenaf or bamboo fiber by preparing laccase crude enzyme solution from Candida tropicalis DK2 is as follows:

[0034] Put 5g of flax, jute, kenaf or bamboo fiber raw stems in a 250ml conical flask, and use flax, jute, kenaf or bamboo fiber raw stems and laccase crude enzyme solution in a mass ratio of 1:20 to pack in conical flasks 100ml liquid, degumming at 40°C, 200rpm shaker for 1 day, 2 days and 4 days respectively; when the degumming time is up, remove the degumming solution, boil in a boiling water bath for 5min, rinse with tap water several times, and Remove bacteria from linen and stop degumming.

[0035] Through the observation of flax, jute, kenaf or bamboo fiber degumming, it was found that after 1 day, the degumming of flax, jute, kenaf or bamboo fiber was obviously dispersed, and the color of the fermentation liquid became dark and turbid; , kenaf or bamboo fibers are dispersed into fibers, and the fermented liquid is ob...

Embodiment 3

[0037] The process of preparing laccase enzyme solution for degumming in flax, jute or kenaf yarn by Candida tropicalis DK2 is as follows:

[0038]Put 5g of flax, jute or kenaf yarn into a 250ml conical flask, and divide flax, jute or kenaf yarn and laccase enzyme solution into the conical flask according to the mass ratio of 1:20, and the liquid volume is 100ml , degumming at 40°C, 200rpm shaker for 1 day, 2 days and 4 days respectively; when the degumming time is up, remove the degumming solution, boil in a boiling water bath for 5min, and rinse with tap water several times to remove flax, jute or kenaf Bacteria on yarn, stop degumming.

[0039] Through the observation of flax, jute or kenaf yarn degumming, it was found that the flax fiber was obviously dispersed after 1 day, and the color of the fermentation liquid became turbid; Days later, the degumming rate of flax reached over 90%.

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Abstract

The invention relates to a method for preparing laccase from candida tropicalis DK2 strains. The method comprises the following steps: (1) transferring the candida tropicalis DK2 strains into a triangular flask containing a basic liquid nutrient medium from a PDA (potato dextrose agar) slant nutrient medium, performing shake culture, filtering fermentation liquor to obtain a mycelium, cleaning the mycelium by using sterilized water, breaking up the mycelium, collecting the mycelium into the sterilized water to prepare a mycelium suspension, and storing for later use; (2) adding an enzyme production nutrient medium into the triangular flask, inoculating the mycelium suspension, performing shake culture, extracting the fermentation liquor regularly during culture, centrifuging, and taking a supernatant as crude enzyme fluid; and (3) centrifuging the crude enzyme fluid, and taking the supernatant to obtain laccase. The method provided by the invention has the characteristics of being simple in process, suitable for large-scale industrial production and the like, and prepared laccase can be used for effectively removing non-cellulose colloids in flax, jute, kenaf or bamboo fiber, has the advantages of high degumming efficiency, short degumming cycle, high fiber dispersion rate, good refined dry flax quality and the like, and has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of biological textiles, in particular to a method for preparing laccase from Candida tropicalis DK2 strain. Background technique [0002] Laccase (Laccase, EC.1.10.3.2) is a copper-containing polyphenol oxidase, which belongs to the same family of ceruloplasmin oxidases as ascorbate oxidase in plants and ceruloplasmin in mammalian plasma. Due to its high redox potential, fungal laccase can act on a wide range of substrates, and can catalyze the oxidation of various phenols and aromatic amines, while reducing molecular oxygen to water. The outstanding catalytic properties of laccase are that its substrates are extensive, the catalytic reaction is complex, and the products produced are environmentally friendly. Therefore, in recent years, the use of fungi to prepare laccase in the degumming of hemp fiber has become more and more extensive. . [0003] There are many kinds of hemp fibers, which are composed of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12R1/74
CPCC12N9/0061C12Y110/03002
Inventor 丁若垚郁崇文张兴群原月梦孔德枝常慧
Owner DONGHUA UNIV
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