Lachnum and method for preparing melanin by liquid fermentation thereof

A technology of Trichodia granulosa and liquid fermentation, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., and can solve the problem of low yield

Inactive Publication Date: 2010-03-17
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

British "Fungus Research" (Mycological Research, 2004, 108 (8): 974-978) once reported that melanin was obtained by culturing in PDB medium for 10 days, and directly participated in the biosynthesis of melanin with L-tyrosine, which was processed by tyrosinase The catalysis of (TYR) makes L-tyrosine hydroxylated to form L-dopa, and oxidized to L-dopaquinone, and L-dopaquinone finally forms melanin throug

Method used

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  • Lachnum and method for preparing melanin by liquid fermentation thereof

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Embodiment 1

[0019] The sporocarp of Lachnum CCTCC M 209193 of the present invention is collected from Huangshan Mountain, Anhui Province, and is separated and purified by a tissue separation method: take fresh Lachnum sporocarp, soak it with 75% alcohol for 10-30s Then soak in 0.1% mercuric chloride solution for 5-8 minutes, and then wash 2-3 times with sterile water, cut the fruiting bodies into 2-4 pieces, inoculate them in PDA medium, and cultivate them at a constant temperature of 23-25°C. After the silk grows, take the bacterial blocks with the same age and shape along the edge of the colony for purification and culture, repeat 4 to 5 times, and obtain the pure culture of Lachnum CCTCC M 209193;

[0020] (1) Liquid Fermentation of Lachnum CCTCC M 209193

[0021] Inoculate the above-mentioned pure culture of Lachnum CCTCC M 209193 on a potato glucose medium (PDA medium) plate for activation. After 7 days of constant temperature cultivation at 25°C, take a piece of bacteria along the e...

Embodiment 2

[0029] The method for obtaining the pure culture of Lachnum (Lachnum) CCTCC M 209193 is the same as in Example 1;

[0030] (1) Liquid Fermentation of Lachnum CCTCC M 209193

[0031] Inoculate the above-mentioned pure culture of Lachnum CCTCC M 209193 on a potato glucose medium plate for activation, and after cultivating at a constant temperature at 23°C for 5 days, use a hole puncher with a diameter of 6 mm to take a piece of bacteria with the same age along the edge of the colony Inoculate in a 250ml Erlenmeyer flask with 50ml potato dextrose liquid fermentation medium, and after cultivating on a shaker at 160r / min for 5 days at 25°C, the seed liquid of Lachnum CCTCC M 209193 was obtained. This Lachnum (Lachnum) CCTCC M 209193 seed solution was inoculated in a 5L automatic control fermenter by 4% of the fermentation medium volume, the fermentation medium filling volume was 3L, and the ventilation rate was 2L / min. After culturing at a rotational speed of 180r / min for 10 days,...

Embodiment 3

[0038] The method for obtaining the pure culture of Lachnum (Lachnum) CCTCC M 209193 is the same as in Example 1;

[0039] (1) Liquid Fermentation of Lachnum CCTCC M 209193

[0040]Inoculate the above-mentioned pure culture of Lachnum CCTCC M 209193 on a potato glucose medium plate for activation, culture at a constant temperature of 25°C for 6 days, and use a hole puncher with a diameter of 6mm to take a piece of bacteria with the same age along the edge of the colony Inoculate in the 250ml Erlenmeyer flask that 60ml potato dextrose liquid fermentation medium is housed, at 23 ℃, with the rotating speed of 200r / min, shaker culture after 3 days, obtain Lachnum (Lachnum) CCTCC M 209193 seed liquid. This Lachnum (Lachnum) CCTCC M 209193 seed solution was inoculated in a 5L automatic control fermenter by 8% of the fermentation medium volume. Rotating speed of 200r / min, after cultivating for 12 days, suction filtration twice, the filtrate is the required fermentation broth. The w...

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Abstract

The invention discloses a lachnum and a method for preparing a melanin by liquid fermentation thereof, and is characterized in that seed liquid which is obtained by the shaking culture after the Lachnum CCTCC M 209193 is activated is inoculated into a fermentation medium to conduct liquid fermentation according to 4-8 percent of the fermentation medium in volume; after the fermentation liquid is extracted with alkaline solution, a precipitation agent is used for precipitation; by the acid hydrolysis of hydrochloric acid, a crude melanin is obtained from a crude extract; then the crude melaninis re-dissolved in ammonia solution; and finally a pure melanin is obtained by using oil ether, chloroform and ethyl acetate for extraction in sequence. Compared with the method which adopts a basic fermentation medium, by adopting the modified fermentation medium and taking the hydrochloric acid methanol solution as a precipitating agent, the yield of the melanin is increased to 3.856g/L from 1.471g/L, the fermentation period is shortened to 8 days from 12 days, and the purity of the crude melanin extract is high, thereby being conducive to the purification and reducing the damage to the structure thereof in the purification process.

Description

technical field [0001] The invention belongs to the technical field of preparing melanin by liquid state fermentation of microorganism Lachnum, and in particular relates to extracting melanin from Lachnum fermentation liquid by using methanolic hydrochloric acid solution as a precipitating agent and adopting an improved fermentation medium to improve Methods of melanin production. Background technique [0002] At present, melanin can be chemically synthesized by oxidation of tyrosine or its derivatives, extracted from animal and plant materials, or synthesized by microorganisms. For example, Chinese Patent Application Nos. 92112247.0 and 91104469.8 respectively disclose a method for preparing melanin by using mammalian animal hair as raw material and black sesame as raw material, leaching with sodium hydroxide solution and using hydrochloric acid as a precipitating agent to precipitate melanin . However, with people's understanding of the safety of melanin synthesized from...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P1/02C09B61/00C12R1/645
Inventor 叶明许庆平陈晓
Owner HEFEI UNIV OF TECH
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