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Method for producing mycelia and exopolysaccharide by marasmius maximus liquid culture

A liquid culture and extracellular polysaccharide technology, applied in the biological field, can solve the problems of small individuals, difficult separation, and destruction of wild resources, and achieve the effects of easy extraction, simple products, and difficult collection

Active Publication Date: 2020-05-22
GUIZHOU ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although large-capped and small-leaf umbrellas are more common in wild conditions, they are not easy to separate due to their small size, and it is difficult to meet market demand only by manual picking. At the same time, excessive picking will also cause damage to wild resources.

Method used

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  • Method for producing mycelia and exopolysaccharide by marasmius maximus liquid culture
  • Method for producing mycelia and exopolysaccharide by marasmius maximus liquid culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Take the triangular flask of 500 milliliters as the fermentation vessel as an example, and carry out the liquid culture of the mycelium of the large cap Pygmycus edulis.

[0031] Inoculate the slant bacteria on the PDA medium, 25 o C for 15 days in the dark, and the grown slant strains were inoculated into the primary liquid medium. The specific formula was as follows: glucose 20 g, peptone 5 g, potassium dihydrogen phosphate 0.5 g, magnesium sulfate 0.5 g, vitamin B1 0.02 g, dissolve the above raw materials in water, adjust the volume to 1L with water, adjust the pH to 6.5 with hydrochloric acid or sodium hydroxide, and pack in 500 mL Erlenmeyer flasks, the volume of each bottle is 250 mL, 25 o Under C, vibrating culture, the shaker speed is 120 rpm, cultivated for 8 days, the mycelial balls are filled with liquid medium, and the average diameter of the mycelial balls is about 2-3 mm, which is used as a first-class strain.

[0032] Inoculate the above-mentioned first-...

Embodiment 2

[0035] Take the triangular flask of 500 milliliters as the fermentation vessel as an example, and carry out the liquid culture of the mycelium of the large cap Pygmycus edulis.

[0036] Inoculate the slant bacteria on the PDA medium, 25 o Cultivate in the dark for 15 days, and inoculate the well-grown slant strains into the primary liquid medium. The formula is as follows: 20 g of glucose, 5 g of peptone, 0.5 g of potassium dihydrogen phosphate, 0.5 g of magnesium sulfate, and 0.02 g of vitamin B1 , the above raw materials were dissolved in water, and the volume was adjusted to 1 L with water, and the pH was adjusted to 6.5 with hydrochloric acid or sodium hydroxide. Packed in 500mL Erlenmeyer flasks, the volume of each bottle is 250mL, 25 o Under C, vibrating culture, the shaker speed is 120 rpm, cultivated for 8 days, the mycelial balls are filled with liquid medium, and the average diameter of the mycelial balls is about 2-3 mm, which is used as a first-class strain.

[0...

Embodiment 3

[0040] Take the triangular flask of 500 milliliters as the fermentation vessel as an example, and carry out the liquid culture of the mycelium of the large cap Pygmycus edulis.

[0041] According to the results of single factor tests such as carbon source and nitrogen source in Example 1 and Example 2, it can be known that cornstarch and peptone are the best carbon source and nitrogen source respectively, which are most conducive to the production of mycelia and have the highest yield. Prepare liquid medium according to these conditions: cornstarch 15 g / L, peptone 5 g / L, vitamin B 1 0.02 g / L, KH 2 PO 4 1 g / L, MgSO 4 0.5 g / L. An orthogonal experiment plan was performed on temperature, shaker speed, and inoculum size of liquid seeds to optimize the best parameters of these three culture conditions. Orthogonal design experiments are shown in Table 1 and Table 2.

[0042] Level temperature °C Speed ​​r / min Inoculation amount % (V / V) 1 20 120 1 2 ...

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Abstract

The invention discloses a method for producing mycelia and exopolysaccharide by marasmius maximus liquid culture. The method mainly comprises the following steps: strain preparation; preparation of aprimary liquid strain; preparation of a secondary liquid strain; production culture medium preparation, inoculation and culture; and collection of mycelia and exopolysaccharide: the mycelia are collected by a centrifugation method or a filtration method, and after reduced pressure concentration of a supernatant obtained through centrifugation or a filtrate obtained by a filtration method, polysaccharide is separated by an alcohol precipitation method and subjected to freeze-drying to prepare polysaccharide freeze-dried powder. The method adopts a marasmius maximus liquid culture production manner and has the advantages of being rapid, efficient and convenient, the product obtained through liquid culture is simpler, easier to extract, separate and purify, the mass production of the marasmius maximus mycelia and exopolysaccharide, so that the purpose of meeting the market requirement is achieved, and the problems that wild resources are difficult to collect and small in quantity are solved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for producing mycelium and exopolysaccharide by liquid culture of large-covered and small-skin umbrellas. Background technique [0002] Microcoria ( Marasmius ) is a class of large fungi with a certain medicinal value, the most famous of which is Anluo Xiaopi umbrella ( Marasmiellus androsaceus ), which is a traditional medicinal fungus and the main raw material of Anluotong. Its mycelium is commonly used among the people in our country to treat bruises, fractures, sciatica, migraine, rheumatic joint pain, etc. It has been confirmed by modern pharmacological experiments that Anluo Xiaopi polysaccharide is one of the important active ingredients of Anluo Xiaopi. It has multiple functions such as improving immune resistance, preventing tumors and eliminating free radicals. The active components such as sugars contained in the mycelium of Anluo Xiaopi umbrella c...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/04C12R1/645
CPCC12N1/14C12P19/04
Inventor 邓春英林群英吴亮亮孙力军姚正颖侯北伟金敬红秦维
Owner GUIZHOU ACAD OF SCI
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