CN1151847A - Chinese glossy ganoderma health beverage - Google Patents

Abstract

The present invention is a health glossy ganoderma beverage which is produced by deep fermentation of strain obtained by separating sporophore of Ganoderma sinense, deactivation and dilution of fermented liquor, and addition of supplement. In the present invention, culture medium of cane sugar is used for industrialized prodn, and the fermented product has high contents of glossy ganodrma polysaccharide, amino acid and trace elements. The said beverage can raise body's immunity.

Description

China's glossy ganoderma health beverage

The present invention relates to non-alcoholic beverage, particularly utilize Chinese glossy ganoderma (Ganoderma sinense Zhao, Xu et Zhang) the made beverage of bacterial classification submerged fermentation technology.

China just is considered as glossy ganoderma the thing of luckiness since ancient times, is referred to as celestial grass.Glossy ganoderma is used for clinical with a long history, and such record is arranged in " (Sheng Nong's herbal classic) ": " purple sesame flavor is sweet, warm.Main deaf, god is protected, beneficial vital essence, hard muscles and bones, good color in sharp joint.Clothes are made light of one's life by commiting suicide for a long time, and are not old, prolong life." therefore, glossy ganoderma is long-tested nourishing and fit keeping function, the valuable ingredient of traditional Chinese medicine of strengthening the body resistance to consolidate the constitution.But glossy ganoderma is natural crude drugs, gathers rather difficulty, and limited amount, and as adopting artificial cultivation, its cultivation cycle is long and labour intensity is big, yields poorly and the cost height, is unfavorable for suitability for industrialized production.Simultaneously, the active ingredient GL-B is than its fructification content Senior Two doubly in the Ganoderma lucidum mycelium.Therefore, explore the glossy ganoderma industrialized preparing process and become the important channel of opening up the glossy ganoderma resource.

Chinese invention patent ublic specification of application CN1105208A discloses the lucidum strain fermented glossy ganoderma series health food of making of a kind of usefulness.The bacterial classification that will separate from ganoderma lucidum fruitbody in this invention utilizes submerged fermentation liquid to make Ganoderma lucidum drink or oral liquid through submerged fermentation, and to filter the filter residue that remains in the Ganoderma lucidum drink be food additives with making, and makes glossy ganoderma food.This invention has effect preferably, but in its used submerged fermentation technology following shortcoming is arranged:

1, selected kind fixing kind not, kind is optimized inadequately, so its product quality is difficult to control, thus the health care of its product also just has difference;

2, the ganoderma polyoses content as active principle is low in the tunning of this invention, generally is not higher than 250mg/100ml;

3, in the deep layer fermenting process of this invention, use with rice and soy meal fluid nutrient medium as main nitrogen and carbon source.The prescription of this culture medium is unsuitable for big industrial production, because the rice as culture medium raw material will pass through boiling, defibrination, the production process complexity, consuming time, consumption worker, soy meal also will be subjected to territorial restrictions simultaneously through complicated processing procedure, buys to be difficult for, cost is also high, thereby hinders the big suitability for industrialized production of glossy ganoderma health beverage;

4, in the filter process of fermentation post processing, it is a kind of waste that the Ganoderma lucidum mycelium in the ganoderma lucidum fermented liquid is filtered out, because also contain some active ingredient in the Ganoderma lucidum mycelium, the mycelia fiber has the effect that promotes digestion simultaneously.

The objective of the invention is to: the glossy ganoderma health beverage that provides a kind of bacterial classification to optimize, utilize the kind China lucidum strain that is filtered out to make the quality height and guaranteed product through submerged fermentation; In submerged fermentation, do not use the rice of fabrication schedule complexity or soy meal raw material as culture medium; The fermentation back maximizes the retention active ingredient without filtration; Acquisition contains GL-B amount height and the low Ganoderma lucidum drink of cost.

The object of the present invention is achieved like this: provide a kind of with ganoderma strain capable through submerged fermentation, make Ganoderma lucidum drink with its zymotic fluid, used glossy ganoderma is adopted Chinese glossy ganoderma CGMCC0270.The strain of this bacterial classification is called 60593 (M) Ganoderma, and this bacterial classification is at China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) application culture presevation, preservation date: on August 23rd, 1996, preserving number CGMCC No.0270.China's this bacterial classification of glossy ganoderma is the bacterial classification that shows unique characteristics that preferably comes out after screening, cultivation, research from nearly hundred kinds of lucidum varieties.This kind ganoderma lucidum fruitbody is characterized as: cap semicircular in shape, subcircular or nearly cochlear, and 2.5～9.5 * 2.2～8cm sometimes can be bigger, thick 0.4～1.2cm, surperficial atropurpureus is to nearly black, puce sometimes, paint-like gloss is arranged, obvious or unconspicuous concentric circular groove of tool and vertical wrinkle; Thin edge or blunt normal approximate is cut a shape, and is homochromy with cap, lighter or be yellowish-brown.The even brown of bacterial context arrives dark brown, thick 0.1～0.3cm, the long 0.3～1cm of tube, brown, dark brown or taupe.The dirty white of hole face, filbert to brown; The mouth of pipe is slightly circular, 5～6 every millimeter.Stem adnation, dorsal part are given birth to or wilfully, and be cylindrical or slightly flat, long 7～19cm, and thick 0.5～1cm, homochromy or color is darker with cap, glossy.

The basidiospore oval, the top navel is protruding or truncate slightly, double wall, outer wall clear, colorless, level and smooth, inwall is filbert, the obvious spinule of tool, the basidiospore size is 9～13.8 * 6.5～8.1 μ m.

The China glossy ganoderma is born in the woods on broad leaf tree or the coniferous tree timber, and the place of production is in Chinese Hainan.

The biological property of China's lucidum strain is: pure white, fine hair shape, initial stage white on solid medium, the later stage yellow that partly shoals.On plating medium, can cover with in general 7 days; At microscopically hyphae colorless, very thin and tool branch, diaphragm and clamp connection; Mycelia is thick during than solid culture when Liquid Culture, and plasm is abundant, and diaphragm and clamp connection are many and clear, are milk yellow bacterium ball to suspend in zymotic fluid;-18 ℃ of following mycelia chaps, plasm is assembled, and still has diaphragm and clamp connection, but form changes death in 4 days.

This bacterial classification is adopted following deep layer fermenting process:

(1) strain inclined plane is cultivated, and is to carry out seed static cultivation in inclined-plane in slant medium, and cultivation temperature is 27～28 ℃, and incubation time generally needs 7 days, and used slant medium is a comprehensive PDA culture medium, and its composition is:

Potato 10～20%, glucose 2～4%, potassium dihydrogen phosphate 0.2～0.4%, magnesium sulfate 0.15～0.3%, vitamin B1 0.1～0.2%, agar 1.8～2%

(2) shaking bottle cultivates: make bacteria suspension with the seed that the inclined-plane is cultivated, in shaking bottle culture medium, under temperature 26-28 ℃, cultivate in rotating speed is 180-200 rev/min shaking table, the used bottle culture medium that shakes is: sucrose 1～3%, dusty yeast 1～2%, potassium dihydrogen phosphate 0.2～0.4%, magnesium sulfate 0.15～0.3%, vitaminB10 .1～0.2%, pH value nature.Wherein, one-level is shaken bottle and was cultivated 5～7 days, and secondary shakes bottle and cultivated 4～5 days.

(3) seed tank culture: the secondary shake-flask seed is inoculated in the seed tank culture liquid, and inoculum concentration is 10%, and cultivation temperature is 26～28 ℃, throughput is 1: 0.7～1 (V/V), 200 rev/mins of speeds of agitator, incubation time 40～45 hours, used seed tank culture liquid is with to shake bottle nutrient solution identical.

(4) fermentation tank culture: use the seed of seeding tank to press 10% inoculum concentration access fermentation tank, 26～28 ℃ of cultivation temperature, throughput are 1: 0.7～1 (V/V), and speed of agitator is 200 rev/mins, incubation time 2～3 days, fermentation tank culture liquid is with to shake bottle nutrient solution identical.

(5) zymotic fluid with fermentation tank culture is heated to 80 ℃, cools off after heating deactivation in 10 minutes, and dilution adds flavoring, promptly makes glossy ganoderma health beverage, or without dilution, adds flavoring, promptly makes health oral liquid of glossy ganoderma.

The present invention has good effect, and the outward appearance of this beverage and oral liquid, fragrance, mouthfeel are fine; Contain abundant high 3～5 times GL-B of ratio prior art, reach 1000～1500mg/100ml, and rich in amino acid, trace element; In beverage, contain the cellulose that abundant Ganoderma lucidum mycelium constitutes, digestion is had facilitation; Through experimental verification, to improving the human immunologic function effect is preferably arranged, also AIDS virus HIV there is certain inhibitory action, inhibiting rate reaches 56.4%; Culture medium adopts SM, and sucrose is easy to buy, and is convenient to big suitability for industrialized production.

The drawing explanation of accompanying drawing:

Fig. 1. the front elevation of Chinese ganoderma lucidum fruitbody;

Fig. 2. the back view of Chinese ganoderma lucidum fruitbody;

Fig. 3. Chinese lucidum strain is cultivated 4 days state in plating medium;

Fig. 4. Chinese lucidum strain is cultivated 7 days state in plating medium;

Fig. 5. at the diaphragm 1 of microscopically mycelia and the state of clamp connection 2;

Fig. 6. 5S RNA (rRNA) PCR of mycelium A of the present invention and fructification B (reaction of archaeal dna polymerase amplification in vitro) amplified production restriction analysis collection of illustrative plates.This collection of illustrative plates is used for classifying on molecular level.

Fig. 7. Chinese glossy ganoderma cultivate on the inclined-plane 4 days 2. with cultivate 7 days states 1.;

Below embodiments of the invention are further described in conjunction with the accompanying drawings:

Fig. 1 is Chinese ganoderma lucidum fruitbody full face, and Fig. 2 is a Chinese ganoderma lucidum fruitbody back side photo, and its fructification feature as previously mentioned.

Fig. 3 is that this bacterial classification was cultivated in plating medium after 4 days, and bacterium colony is the form that radioactivity is launched; Fig. 4 is that this bacterial classification is cultivated after 7 days in plating medium, and bacterium colony covers with the state of media surface.Fig. 5 is the diaphragm 1 between hyphal cell and the electron micrograph of clamp connection 2.More than three width of cloth figure biological property of Chinese lucidum strain is described, particular content is as previously mentioned.

Fig. 6 is 5S RNA (rRNA) PCR (reaction of archaeal dna polymerase amplification in vitro) the amplified production restriction analysis collection of illustrative plates of mycelium A of the present invention and fructification B.This collection of illustrative plates is used for classifying wherein that A is the mycelium pcr amplification product on molecular level, B is the fructification pcr amplification product, and the explanation of this collection of illustrative plates: the mycelia of the bacterial classification that adopts in the submerged fermentation technology of the present invention and fructification are to belong to same bacterial strain.

The compound method of culture of strains process and beverage:

(1) cultivation of the inclined-plane of bacterial classification is to carry out seed static cultivation in inclined-plane in slant medium, and cultivation temperature is 27～28 ℃, and incubation time generally needs 7 days, and used slant medium is a comprehensive PDA culture medium, and its composition is:

Potato 10～20%, glucose 2～4%, potassium dihydrogen phosphate 0.2～0.4%, magnesium sulfate 0.15～0.3%, vitamin B1 0.1～0.2%, agar 1.8～2%

To be Chinese lucidum strain on the inclined-plane cultivate Fig. 7 2. reached 7 days states 1. in 4 days, and wherein: after 2. cultivating 4 days, bacterium colony is that the center is radial outside expansion with the inoculation position.1. after cultivating 7 days, bacterium colony covers with media surface.

(2) shaking bottle cultivates: make bacteria suspension with the seed that the inclined-plane is cultivated, in shaking bottle nutrient solution, under 26～28 ℃ of the temperature, cultivate in rotating speed is 180～200 rev/mins shaking table, the used bottle nutrient solution that shakes is: sucrose 1～3%, dusty yeast 1～2%, potassium dihydrogen phosphate 0.2～0.4%, magnesium sulfate 0.15～0.3%, vitaminB10 .1～0.2%, pH value nature.Wherein, one-level is shaken bottle and was cultivated 5～7 days, and secondary shakes bottle and cultivated 4～5 days.The result is as shown in table 1 for 250ml triangular flask shake flask fermentation, as seen wherein contains abundant GL-B.

Table 1 250ml triangular flask shake flask fermentation result

Lot number	1	?2	?3
				Stop pH value	4.0	3.5	?3.8
Final volume (ml)	31	110	?138
				Viscosity (second)	38	18.5	?11.4
GL-B (mg/ml) in the cell	4.2	1.65	?0.54
				Extracellular GL-B (mg/ml)	15.2	18.52	?20.08
Intracellular polyse+exocellular polysaccharide (mg/ml)	19.4	20.17	?20.62

(3) seed tank culture: be inoculated in the seed tank culture liquid with the secondary shake-flask seed liquid, inoculum concentration is 10%, and temperature is 26～28 ℃, throughput is 1: 0.7～1 (V/V), 200 rev/mins of speeds of agitator, incubation time 40～45 hours, used seed tank culture liquid is with to shake bottle nutrient solution identical.

(4) fermentation tank culture: the seed of seeding tank is inoculated in the fermentation tank in 10% ratio, 26～28 ℃ of cultivation temperature, throughput are 1: 0.7～1 (V/V), and speed of agitator is 200 rev/mins, incubation time 2～3 days, fermentation tank culture liquid is with to shake bottle nutrient solution identical.

(5) zymotic fluid with fermentation tank culture is heated to 80 ℃, cools off after heating deactivation in 10 minutes, and dilution adds flavoring, promptly makes glossy ganoderma health beverage.

The compound method of beverage of the present invention is as follows: above-mentioned Chinese ganoderma lucidum fermented liquid is added 4～5 times of water, add 2～5% sucrose or albumen sugar, add 1～2% lactic acid, promptly become Ganoderma lucidum drink after stirring.Original fermentation liquor can not added water adding flavoring yet and make health care oral liquid.

Ganoderma lucidum drink outward appearance of the present invention, fragrance and mouthfeel are all good, contain abundant GL-B, amino acid and trace element; Contain the cellulose that abundant Ganoderma lucidum mycelium constitutes in the beverage, can promote digestion; Through experimental verification, can improve the human immunologic function, the inhibiting rate of AIDS virus HIV is reached 56.4%.Table 2 is the amino acid analysis results that done by the Beijing City Nutrient Source Inst, and the instrument of use is the 835-50 of a Hitachi type chromatograph.Tryptophan is measured with the Hydrolyze method of the NaOH of 4.2N, handles with the performic oxidation method and measures cystine, and all the other amino acid are measured with the HCl hydrolysis of 6N, and last column volume is 50 microlitres, and table 2 explanation amino acid content is abundant; Table 3 is by the just micro-analysis result of being done in Beijing City Nutrient Source Inst: except that table is gone up listed abundant trace element, do not detect lead and arsenic as harmful element, and therefore, useful and harmless to health.In addition, through life science institute of Nankai University bioengineering bacterial classification submerged fermentation product 1025B of the present invention is detected three kinds of methods with plasomidum calibrating, cell survival rate calibrating and Luc vigor detect and confirm: the submerged fermentation product of bacterial classification of the present invention has the effect that certain inhibition plasomidum forms, do over against confirming that according to medicine the submerged fermentation product 1025b of bacterial classification of the present invention has certain inhibitory action to AIDS virus HIV with AZT, inhibiting rate is 56.4%.

Table 2 amino acid analysis result (mg/100mg sample)

Asparatate	0.0250	Threonine	0.0075
				Serine	0.0076	Glutamic acid	0.0409
Glycine	0.0077	Alanine	0.0066
				Valine	0.0047	Methionine	0.0010
Isoleucine	0.0033	Leucine	0.0045
				Tyrosine	0.0023	Phenylalanine	0.0032
Lysine	0.0073	Ammonia	0.0083
				Histidine	0.0023	Arginine	0.0043
Proline	0.0055	Tryptophan	0.0004
				Cystine	0.0029
Summation	0.1453

Table 3 trace element analysis result

Element	Potassium K	Sodium Na	Magnesium Mg	Iron Fe
					Content	????0.18％	????623mg/kg	????317mg/kg	????0.31mg/kg
Element	Copper Cu	Manganese Mn	????Zn
					Content	????0.046ppm	????0.041ppm	????1.3ppm

Claims (1)

1, Chinese glossy ganoderma health beverage through submerged fermentation, is made Ganoderma lucidum drink with its zymotic fluid with ganoderma strain capable, it is characterized in that:

Adopt Chinese glossy ganoderma (Ganoderma sinense Zhao, Xu et Zhang) CGMCC No.0270, the cap semicircular in shape of this bacterial classification fructification, subcircular or nearly cochlear, 2.5～9.5 * 2.2～8cm sometimes can be bigger, thick 0.4～1.2cm, the surface atropurpureus is to nearly black, sometimes puce has paint-like gloss, obvious or unconspicuous concentric circular groove of tool and vertical wrinkle; Thin edge or blunt normal approximate is cut a shape, and is homochromy with cap, lighter or be yellowish-brown; The even brown of bacterial context arrives dark brown, thick 0.1～0.3cm, the long 0.3～1cm of tube, brown, dark brown or taupe; The dirty white of hole face, filbert to brown; The mouth of pipe is slightly circular, 5～6 every millimeter; Stem adnation, dorsal part are given birth to or wilfully, and be cylindrical or slightly flat, long 7～19cm, and thick 0.5～1cm, homochromy or color is darker with cap, glossy; The basidiospore oval, the top navel is protruding or truncate slightly, double wall, outer wall clear, colorless, level and smooth, inwall is filbert, the obvious spinule of tool, the basidiospore size is 9～13.8 * 6.5～8.1 μ m; The China glossy ganoderma is born in the woods on broad leaf tree or the coniferous tree timber, and the place of production is in Chinese Hainan; The biological property of China's lucidum strain is: pure white, fine hair shape, initial stage white on solid medium, the later stage yellow that partly shoals; On plating medium, can cover with in general 7 days; At microscopically hyphae colorless, very thin and tool branch, diaphragm and clamp connection; Mycelia is thick during than solid culture when Liquid Culture, and plasm is abundant, and diaphragm and clamp connection are many and clear, are milk yellow bacterium ball to suspend in zymotic fluid;-18 ℃ of following mycelia chaps, plasm is assembled, and still has diaphragm and clamp connection, but form changes death in 4 days;

This bacterial classification adopts following deep layer fermenting process:

(1) strain inclined plane is cultivated, and is to carry out seed static cultivation in inclined-plane in slant medium, and cultivation temperature is 27～28 ℃, and incubation time generally needs 7 days, and used slant medium is a comprehensive PDA culture medium, and its composition is:

Potato 10～20%, glucose 2～4%, potassium dihydrogen phosphate 0.2～0.4%, magnesium sulfate 0.15～0.3%, vitaminB10 .1～0.2%, agar 1.8～2%

(2) shaking bottle cultivates: make bacteria suspension with the seed that the inclined-plane is cultivated, in shaking bottle nutrient solution, under temperature 26～28C, cultivate in rotating speed is 180～200 rev/mins shaking table, the used bottle nutrient solution that shakes is: sucrose 1～3%, dusty yeast 1～2%, potassium dihydrogen phosphate 0.2～0.4%, magnesium sulfate 0.15～0.3%, vitaminB10 .1～0.2%, pH value nature; Wherein, one-level is shaken bottle and was cultivated 5～7 days, and secondary shakes bottle and cultivated 4～5 days;

(3) seed tank culture: the secondary shake-flask seed is inoculated in the seed tank culture liquid, and inoculum concentration is 10%, and cultivation temperature is 26～28 ℃, throughput is 1: 0.7～1 (V/V), 200 rev/mins of speeds of agitator, incubation time 40～45 hours, used seed tank culture liquid is with to shake bottle nutrient solution identical;

(4) fermentation tank culture: the seed of seeding tank is inserted fermentation tank by 10% inoculum concentration, cultivation temperature 26～28C, throughput are 1: 0.7～1 (V/V), and speed of agitator is 200 rev/mins, incubation time 2～3 days, fermentation tank culture liquid is with to shake bottle nutrient solution identical;

(5) zymotic fluid with fermentation tank culture is heated to 80 ℃, cools off after heating deactivation in 10 minutes, and dilution adds flavoring, promptly makes glossy ganoderma health beverage.

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