KR20140064514A - Process for mass production of sparassis crispa mycelia by fed-batch culture - Google Patents

Abstract

 The present invention relates to a method for mass production of mycelia of flowering mushroom by cultivating mycelia of Zygomycorrhizae using nutrients of a citrus peel and a non-matured field, which are lung biomass resources, as well as solving environmental pollution by decomposing citrus industry byproducts, Food and health functional food raw materials.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a process for mass production of sparassis crispa mycelia by fed-

The present invention relates to a mass production method using fed-batch culture of mycelia of P. japonica using a citrus industrial by-product as a culture substrate.

Normally, the mushroom (name: Sparassis crispa Wulf. ex Fr.) is a mushroom of mushroom, mushroom, mushroom, mushroom, and mushroom. In English, it is called cauliflower mushroom or crested sparassis, Japanese name is Hanabiratake, and Chinese name is called sphere.

When the fruiting body of mushroom is matured, the whole is 95-225x110-285mm, and it is somewhat round, and small petal-shaped leaves gather to form flower cabbage-seaweed. It is 25-55x 28-46mm in length, short, stout, hard, repeatedly divided upwards to form a short basal number, the basin flattened, and thin, with a petal-shaped cap. The upper surface of the umbrella is smooth, white-pale yellow, but after growing it is ocher. The follicle layer is on the back (bottom) or outside of each small god, smooth, initially light yellow or yellowish when grown and becomes brown when homeless. The tissue is thin, elastic, flexible, fleshy, white.

The results of the analysis of the mushroom component of Zygomycetes mushroom in the immunology classroom of the Department of Pharmacy, Faculty of Pharmaceutical Sciences, Tokyo University of Medicine in March 2001 showed that 43.6% (43.6g in 100g) of beta-glucan was contained in the dried mushroom. The mushroom of this kind can be said to be a beta-glucan mass. The beta-glucan has been found to regulate the immune system of the body as a key ingredient for increasing the immunity of the human body, and to control cancer, hypertension and diabetes.

In addition to the excellent anticancer effect, the above-mentioned Zygomycetes mushroom not only lowers the blood pressure and returns the blood glucose level to normal but also has a hematopoietic effect, but the amount that can be caught in the natural state is extremely small and efforts for artificial cultivation continue come.

Japanese Patent Laid-Open Publication No. 11-56098 discloses a method for cultivating a Japanese mushroom by using a medium supplemented with a nutrient source after extracting hot water from a larch and removing water components. Korean Patent Publication No. 2002-48337, The method of artificial cultivation of mushroom includes adding activated carbon and active calcium to a medium containing a mixture of larch and light leaf and sterilizing a medium containing a commonly used nitrogen source at a high temperature and a high pressure, Has been disclosed as an artificial cultivation method.

In addition to the above artificial cultivation method of the mushroom as described above, Korean Patent Publication No. 2004-0071926 'Method of Culturing Mycelium of Zygomyceta mushroom' is not artificial cultivation for producing fruiting body of mushroom, but a simple method using mycelium of Mushroom A liquid culture method has been disclosed.

 About 20% of Jeju Island's total citrus production is used as mandarin orange. The citrus fruit produced in this citrus processing industry is estimated to be 100,000 tons per year. Currently, it is pointed out that it is one of the most important causes of environmental pollution in Cheju Island, which is the largest tourist spot in Korea, because it is difficult to completely treat citrus peel. Studies on the use of citrus peel for feed and fertilizer have been continuously carried out, but there is a great difficulty in industrialization due to economical efficiency and inefficient production thereof. Currently, most of the citrus peel is marine dumping, but since July 07 when the London Treaty comes into force, the ban on marine dumping of citrus peel is completely banned. Therefore, it is not an exaggeration to say that the establishment of the treatment method of citrus peel is the most influential factor in the development of the citrus processing industry.

The water content of citrus juice, which is a waste of citrus juice processing, is 82%, which is a major constraint factor for the treatment of by - products and useful resources. Citrus juice leaves contain 10-15% of pectin and 1-3% of flavonoids in dry weight. Pectin is a complex polysaccharide mainly present in the middle lamella of a plant, and refers to a substance that binds and binds cells. It is widely used as a gelling agent and has effects such as prevention of hypertension, arteriosclerosis and angina, removal of heavy metals, anti-radiation, lowering of cholesterol in blood, proliferation of useful microorganisms in the intestines, and strengthening of the immune system. There are about 60 kinds of citrus flavonoids, and common flavonoids such as rutin and theosmin, and flavonoids specific to citrus such as neranine and hesperidin, novirretin and benzeretin. It has been found that there are many excellent natural materials with excellent physiological activity among flavonoids. In particular, antioxidant activity, prevention of circulatory diseases, anti-inflammatory, antibacterial, antiviral. Immune enhancement, liver disease improvement effect, anti-cancer function has been revealed various biocontrol functions. Using these characteristics, it is possible to develop functional foods and medicines for prevention and treatment of diseases of adult diseases such as hypertension, arteriosclerosis, angina pectoris, and diabetes.

It is an object of the present invention to provide a method for producing a beta-glucan high-content Zygomycetes mushroom mycelium (hereinafter, referred to as " Zygomycetes ") through a fed-batch culture using a citrus industrial by-product, And a method for producing the same.

In one aspect, the present invention relates to a method for mass production of a mycelium of mushroom mycelia capable of reducing the unit cost of culture and helping immunity by using citrus peel and immature part, which are difficult to dispose of, as a carbon source. As a means of solving the problem, the citrus industrial by-product was used as a carbon source in the cultivation of the mycelia of the mycelial growth of mycelia, and a concentrate medium containing glucose and yeast extracts was supplied intermittently or continuously during the culture, The specific growth rate was investigated to establish the optimal fermentation process.

Using the citrus industrial by-product as a culture substrate, we have developed a process for the oil-price cultivation of mycelia of Zanthoxylum mushroom, and have specifically developed a large amount of mycelia of Zanthoxylum mushroom which can be used not only as a raw material for health functional food, And a method for cultivation for production.

Fig. 1 is a graph showing an increase in mycelia growth of mycelia of leaves from intermittent feeding method
FIG. 2 is a graph showing the growth of mycelium of Mytilus edulifera by continuous feeding method

<Example 1> Liquid culture method of Mycelia

(1) germinating the spores of Zinnia mushroom to obtain mycelium;

Mycelia of Zygomyksis mushroom were obtained by germination of spores obtained from the fruiting body of Zygomy mushroom collected in the wild. Yeast - Potato starch - It is obtained by spreading on glucose agar (3g of yeast extract, 10g of potato starch, 10g of glucose, 15g of agar, 1ℓ of distilled water) for 15 days at 23˚C. On the other hand, the mycelium is sliced on a test tube containing yeast-potato starch-glucose agar medium, stored at 4 ° C, and subcultured every month.

(2) culturing the mycelium in a liquid medium;

Mycelia grown in the slope medium are aseptically collected and inoculated into the liquid medium at 10% (v / v). The mycelium productivity is good when the medium is adjusted to 3% of glucose, 0.5% of yeast extract, 0.2% of malt extract, 1% of potassium monophosphate and 0.04% of ferrous sulfate to pH 4.5. Liquid culture of mycelium is cultured in a fermenter at 23 ° C, stirring speed of 200 rpm, aeration amount of 1 vvm for 7 days.

(3) a step of analyzing the mycelia concentration of P. japonicus;

   In order to compensate for the disadvantage that the cell concentration of the actual production strain can not be measured with a simple dry cell mass due to the citrus peel and immature and insoluble matter derived from the immature cell, the present invention analyzes the content of ergosterol existing only in fungi, Respectively. The conditions for HPLC analysis of ergosterol are as follows.

 Column: ODS-2 column

 Mobile phase: MeOH: Acetonitrile (35:65)

 flow rate: 1.2 ml / min

 UV wavelength: 282 nm

The correlation between the ergosterol and the mycelium of P. japonica is shown in Fig.

<Example 2> Fed-batch culture through supply of carbon source and nitrogen source for mass production of mycelia of Zanthoxylum mushroom

 In order to increase the yield and shorten the cultivation period of mycelium of Zygomyx mushroom, the nutrient culture with the nitrogen source, which is a carbon source, was supplied intermittently and continuously. The initial glucose concentration in the medium was added at 10 g / L and the culture was supplemented with 10 g / L of glucose and 5 g / L of yeast extract in an intermittent and continuous manner according to depletion of carbon source and nitrogen source. In the fed-batch culture using the intermittent feeding method, glucose and yeast extract were fed intermittently at intervals of 24 hours from the third culture day to the fifth culture day as shown in Fig. In the fed-batch culture using the continuous feeding method, as shown in Fig. 2, the glucose and yeast extract were supplied so that the glucose concentration was maintained at 10 g / L in consideration of the growth of the mycelium from the third culture day to the fifth culture day.

 As a result, the final dried mycelium amount was 20 g / L and the final dried mycelial amount was 25 g / L by continuous feeding method. The yield was increased by 42% and 78%, respectively, in case of fed-batch cultivation by intermittent feeding method and continuous feeding method, compared with 14 g / L of dry mycelium obtained by batch culture with initial glucose concentration of 20 g / L .

None.

Claims (2)

A culture medium containing 5 to 100 g / L of glucose, 1 to 20 g / L of peptone and 1 to 20 g / L of yeast extract was added to the fermenter, the temperature was maintained at 25 ° C, The initial concentration of glucose is 10 g / L, and 10 g / L of glucose is intermittently supplied at intervals of 24 hours or the glucose concentration is kept constant at 10 g / L Wherein the mycelial growth of mycelium of P. japonica is continued while being continuously supplied. Wherein the supplemental feed is intermittently or continuously fed with glucose alone or a mixture of 10 g / L of glucose and 5 g / L of yeast extract.

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