CN106342092B - The schizophyllum commune fermentate of resistance to anoxic and fermentation process thereof and culture medium - Google Patents
The schizophyllum commune fermentate of resistance to anoxic and fermentation process thereof and culture mediumInfo
- Publication number
- CN106342092B CN106342092B CN200410028929.1A CN200410028929A CN106342092B CN 106342092 B CN106342092 B CN 106342092B CN 200410028929 A CN200410028929 A CN 200410028929A CN 106342092 B CN106342092 B CN 106342092B
- Authority
- CN
- China
- Prior art keywords
- anoxic
- fermentation
- resistance
- fermentate
- culture medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
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Abstract
The invention discloses a kind of produce taking schizophyllum commune as the starting strain fermentate of resistance to anoxic and fermentation method for producing thereof and culture medium. The primary raw material of culture medium provided by the invention comprises the powder of seed body any in highland barley, barley, wheat and maize cereal section crop, seed body, the powder juice of seed body, sprout, the bud powder of seed body or the bud juice of seed body of seed body; Fermentation process can be liquid aerobic fementation, liquid static fermentation or solid-state static fermentation. The fermentate of resistance to anoxic provided by the present invention can improve body hypoxemia tolerance, be conducive to reduce and alleviate a series of anoxic symptoms such as altitude sickness, be particularly suitable for that radical or resident high parent unit is trained under anaerobic environment, patrol, manoeuvre and do to use wartime, also be suitable for using at the special dimension such as Aeronautics and Astronautics and diving, can also be used for field or the crowds such as sports, highland tour and plateau operation.
Description
Technical field
The present invention relates to a kind of product of resistance to anoxic and production method thereof and culture medium, particularly the resistance to anoxic of a kind of schizophyllum communeFermentate and fermentation method for producing thereof and culture medium.
Background technology
Anoxic is extensively present in the mankind's live and work, is especially present in altitude environment, high-altitude or strenuous exercise's mistakeCheng Zhong. In the time that normal human tissue cell can not get the necessary oxygen of metabolic activity, just can produce air hunger, be called for shortAnoxic (Hypoxia, Anoxia, Oxygen deficiency). This is because the insufficiency of intake of oxygen is to supply with bodyRequired, cause alveolar oxygen partial pressure and blood oxygen saturation to reduce, histocyte can not obtain required oxygen from blood just to carry outNormal oxidative metabolism and the series of symptoms that occurs. These symptoms generally comprise: palpitating speed, dry, lip be blue,Dizzy, headache, further can occur feeling sick, vomiting, appetite decline, abdominal distension, diarrhoea, palpitaition, very short of breathTo oedema so that malaise, insomnia, stupor etc. Therefore military affairs, physical culture, Aero-Space, underwater operation, tourismMore and more pay attention to the research to anoxic etc. each field.
In daily life, work and sports, body energy supply mainly contains Aerobic energy supply and anaerobic energy supply. Body is in playIn strong motion, first energy substance produces pyruvic acid and NADH by glycolysis, then in aerobic metabolism process, passes throughThe energy supply of tricarboxylic acid cycle complete oxidation, each molecule glucose produces 36-38 molecule ATP, and accessory substance is water and dioxyChange carbon. Anaerobic metabolism production capacity process only produces the ATP of 2 molecules, and accessory substance is lactic acid. The H+ that lactic acid dissociates makesThe pH value of muscle declines, and along with the increase of lactic acid content in muscle, causes fatigue to increase the weight of.
In the low environment of the oxygen saturation such as plateau, high-altitude, anoxic is a very general problem. General of short duration and journeySpend light anoxic and can cause the light symptoms of some human bodies, as dizziness, headache etc.; If for a long time or sharply anoxic,Because body oxidative metabolism is obstructed, power generation deficiency is even exhausted, can cause serious dysfunction or pathologicChange, occur the disease of many correlations, (for example pulmonary edema, encephaledema, HAHD, erythrocyte increaseDisease, high altitude hypotension disease, high altitude hypertension disease etc.), and may finally cause the termination of vital movement.
Anoxic can exert an influence to every physiological function of body, is embodied in: (1) nervous system: nerveous systemUniting, particularly cerebral cortex is the most responsive to anoxic. The impulsion that when slight hypoxia, chemocepter stimulates anoxic can be madeA series of adaptability compensatory responses (palpitate quickly, blood flow accelerate); When malhypoxia, nervous excitation process weakens,The regulating action of some organs relevant with organism adaptation is strengthened; When severe depletion of oxygen, excitement and conditioned reflex weaken gradually,Cause physiological function insufficiency of accommodation. (2) digestive system: under anoxia condition, animal or human reduces the picked-up of food,Appetite declines, feels sick, the secretory volume of vomiting, all kinds of glandula digestives all declines to some extent, digestive function weakness, stomach emptyingTime lengthening, the activity of intestines is suppressed, and tension force weakens, and peristaltic velocity and amplitude reduce, and have indigestion more. (3)Respiratory system: when anoxic starts, respiratory rate is accelerated, and lung capacity strengthens, and pulmonary ventilation volume increases, partial pressure of oxygen in alveolarIncrease, these changes are conducive to organizing oxygen supply. When malhypoxia, pulmonary surfactant reduces, elastical retraction powerDecline. Excessive expiration can make carbon dioxide discharge too much, and blood pH raises, and occurs alkalemia. (4) cyclic systemSystem: when anoxic, first occur palpitating speed, it adapts to meaning is to try hard to increase cardiac output, is beneficial to and organizes oxygen supply;When malhypoxia, the rhythm of the heart increases but stroke output is the decline of carrying out property gradually; When severe depletion of oxygen, the rhythm of the heart slows down; The utmost pointWhen degree anoxic, the rhythm of the heart can increase again, or with cardiac arrhythmia, extremely slows down subsequently, until pulse stops. In urgencyProperty when anoxic, the CBF of brain increases to improve blood fortune oxygen ability, but continuation increase may cause encephaledema to occur.(5) urinary system: when anaerobic condition, renal blood flow, renal plasma flow all reduce, and anoxic causes after pituitaryThe secretion of leaf antidiuretin increases, therefore there is oliguresis phenomenon, the time is longer, and physical load is larger, occurs the chance of albumenMore. (6) internal system: at the anoxic initial stage, sympathicoadrenal system excitability strengthens, and shows as adrenalineIncrease with noradrenaline concentration, in urine, catecholamine is discharged and is increased, and is tending towards gradually normal later.
Anoxic also can exert an influence to the metabolism of body, concrete manifestation in the following areas: (1) carbohydrateMetabolism: when anoxic, digestive function declines, and stomach tension force reduces, and emptying time is long, and digestive juice secretion amount reduces, appetiteDecline with thirsty sense, can not meet the needs on organism physiology. Meanwhile, glucose absorption slows down, and blood sugar reduces, sugarFormer decomposition is strengthened, and synzyme vigor declines, and gluconeogenesis is obstructed, and glycogen storage level reduces. In addition, under aerobic metabolismFall, anerobic glycolysis is strengthened, and blood lactic acid content raises and accumulation in vivo. (2) protein metabolism: when anoxic,Reduce containing nitrogen compound intake, protein and amino acid decompose to be strengthened, and the discharge rate of nitrogen increases; Protein is syntheticRate declines; In serum, essential amino acid/nonessential amino acid ratio declines. In the time of acute anoxia, high-protein dietCan reduce the endurance to anoxic, and higher nervous activity is had to destruction. (3) lipid metabolism: when anoxic, bloodSlurry free fatty, triglycerides, cholesterol, phosphatide etc. all increase, and may be because lipoprotein lipase vigor subtractsWeak and hormone-sensitive lipase vigor strengthens, and lipolysis is greater than synthetic; Fat reserves reduce and fitting acute anoxiaShould be ineffective, may be because fat covers erythrocyte surface, affect the combination of hemoglobin and oxygen and carry, cause movingThe arteries and veins oxygen content of blood and blood oxygen concentration decline. (4) power and water solution matter metabolism: when acute anoxia, water metabolism presents negative flatWeighing apparatus, electrolyte metabolism gets muddled, and body fluid enters in cell from extracellular, and extracellular fluid reduces, and intracellular fluid increases,Edema. Due to the reduction of blood oxygen dividing potential drop and partial pressure of carbon dioxide, cause haemoconcentration to rise and alkaline reserve simultaneouslyReduce. (5) metabolism of trace elements: under anoxia condition, trace element in vivo be distributed with obvious change, canThe trace element of employing is released into blood, forms recovery organization and breathes and eliminate the necessary enzyme of metabolic disorder productMolecule and other metalloprotein compounds. So trace such as Cu, Zn, Fe in blood, brain, kidney, liver and cardiac muscleElement increases, and the activity of the metalloenzyme such as cytochrome oxidase, catalase raises. But in body trace element notIn the situation of disconnected consumption and supply deficiency, metabolism negative balance will cause these constituent contents in blood and tissue to reduce.
In addition, anoxic also has some impacts to the metabolism of vitamin and nucleotides. Therefore, countries in the world antagonism anoxicResearch attach great importance to, wherein with the tool representative of American and Britain, day, seal, Russia and China.
At present, the major measure of anti-anoxic is pharmaceutical means, and anti-anoxic medicine can be divided into Chinese medicine and the large class of Western medicine two. (1)Chinese medicine: China's traditional Chinese medicine circle is according to the theory of traditional Chinese medical science organic conception and diagnosis and treatment, from supplementing qi and nourishing yin, blood circulation promoting and enriching,The angle of the principles of reatment such as purging the lung of pathogenic fire Li Shui is set out, and carry out the screening operation of resistance to anoxic medicine, and what filter out has a resistance to anoxicThe cmm of simple prescription of effect has: ginseng, Radix Codonopsis, wilsonii, Radix Angelicae Sinensis, cultivated land, the root of straight ladybell, rhodiola root, Poria cocos etc.; MultipleFang You: compound codonopsis tablet, add taste Shengmai San etc. (2) Western medicine: Western medicine kind has amphetamine, chlorpromazine, An ShuLead to etc.
Improve the anoxia tolerance of body by food nutrition measure, particularly prepare resistance to anoxic with microbial fermentation processesThe research report of food seldom. Compared with pharmaceutical means, prepare anoxia-resistant food with microbial fermentation processes and have manyAdvantage, it not only have raw material sources extensively, stable, and can overcome the cost that may exist in chemical synthesis high andTo the pollution problem of environment, avoid Chinese herbal medicine in artificial cultivation process, to be subject to soil, weather, moisture, disease and pest, artificial barThe impact of part etc. Meanwhile, can cultivate screening by microorganism and obtain high-yield strains, fermentation condition is easily controlled, product qualityGuaranteed, be easy to the advantages such as large-scale industrial production. Therefore, anoxia-resistant food has safety, raw material sources are extensive, rawProduce the advantages such as cost is low, production technology is simple.
The fermentation of microorganism has the method such as solid state fermentation, liquid fermentation method. Liquid fermentation method has and comprises liquid static trainingSupport and liquid culture under aeration.
The culture medium of solid state fermentation is solid-state, and sweat adopts static cultivation, and the following process of fermentation ends is simple, GuState fermentation production efficiency is high, and technique is simple, operate extensive, less energy consumption, waste liquid is few, product separation is easier to, produceCost is lower, has good economy and environment benefit.
It is that bacterial classification is received on aseptic liquid culture medium that liquid ventilation is cultivated, and keeps uniform temperature, in incubationVentilate and stir. Liquid ventilation is cultivated has the automation of being convenient to control and large-scale culture, and labour intensity is little, butTo production equipment require high.
Liquid static culture is that bacterial classification is received on aseptic fluid nutrient medium, leaves standstill at a certain temperature cultivationMicrobial culture method. Under many circumstances, aerobic bacteria thalline can form one or more microorganism on liquid surfaceFilm (bunch). According to the difference of thalline and product characteristics, after certain hour is cultivated, the metabolite that thalline producesBe diffused in culture medium, or be trapped in microbial cell, or both have. The liquid layer of product output and culture mediumBetween thickness and surface area, be certain dependency relation. It is large that liquid static culture method has labour intensity, and floor space is largeEtc. shortcoming, but its advantage is simple, and small investment is conducive to shorten fermentation time, do not need to stir and ventilate,Production equipment is required to the advantages such as low, the saving energy.
Schizophyllum is in mycota (Fungi), Basidiomycota (Basidiomycophyta), Hymenomycetes(Hymenomycetes), Holobasidiomycetidae ((Holobasidiomycetidae), Aphyllophorales(Aphyllophorales), schizophyllum commune section (Schizophyllaceae), Schizophyllum (Schizophyllum),Schizophyllum commune (Schizophyllum commune). In this mushroom entity and zymotic fluid thereof, all contain abundant immunityPolysaccharide, protein, biologically active L-type amino acid, necessary 8 seed amino acids of human body, and human body institute is essentialTrace elements of Ca, Co, Cr, Fe, Cu, Mn, Zn etc. When mycelia submerged fermentation, produce organic acid, celluloseEnzyme, malic acid, regulator heteroauxin. Can be widely used in food industry, medical and health, biochemistry aspect.
Summary of the invention
The object of the present invention is to provide a kind of fermentate of resistance to anoxic culture medium of producing taking schizophyllum commune as starting strain.
This culture medium, with any sprout, seed body in the cereal section crops such as highland barley, barley, wheat and maizeBud powder or the bud juice of seed body be primary raw material. Also comprise nitrogenous source, nitrogenous source be peptone, yeast extract, ammonium chloride,The combination of one or more in sulfuric acid amine and ammonium nitrate.
In this culture medium, as the preferably highland barley of cereal section crop of main former section; Be preferably 10-14 ° of Brix highland barley budJuice and 0-0.5%NH4Cl。
Another object of the present invention is to provide a kind of fermentate of resistance to anoxic.
This fermentate of resistance to anoxic, obtains by schizophyllum commune (Schizophyllum commune) fermentation mediumTunning. Described tunning is liquid state fermentation product or product by solid-state fermentation, and wherein liquid state fermentation product is fermentationIn liquid, remove the liquid product after thalline, its content of reducing sugar 15-40g/L, total sugar content 30-70g/L or thalline are rawThing amount 10-30g/L; Described product by solid-state fermentation is that the liquid product of removing in zymotic fluid after thalline obtains through post processingThe solid product that obtains of solid product, schizophyllum commune fermentation solid medium or the powder of this solid product.
Still a further object of the present invention is to provide a kind of method of producing the fermentate of resistance to anoxic taking schizophyllum commune as starting strain.
The method comprises taking schizophyllum commune (Schizophyllum commune) as starting strain, fermentation medium,To the step of the fermentate of resistance to anoxic, comprise successively slant strains cultivation, level liquid seed culture, secondary liquid seedsThe process of cultivation and fermented and cultured, wherein: secondary liquid seed culture medium is by 12 ° of Brix highland barley bud juice and 0.5%NH4ClComposition. Wherein said fermentation is liquid aerobic fementation, and culture medium is 10-14 ° of Brix highland barley bud juice and 0-0.5%NH4Cl，Condition of culture is fermentation temperature 15-35 DEG C, air blow and agitation when fermentation, and fermentation time is 3-6D; Described zymotic fluid can beLiquid static fermentation, culture medium is highland barley flour juice or highland barley bud juice, condition of culture is fermentation temperature 15-35 DEG C, fermentationTime zymotic fluid leave standstill, fermentation time is 3-6D; Described fermentation also can be solid-state static fermentation, and culture medium is highland barley seedBody, highland barley flour, highland barley bud powder, condition of culture is fermentation temperature 15-35 DEG C, leaves standstill and cultivates, fermentation time when fermentationFor 3-6D.
In the method, also comprise the step that fermentate post processing is formed to solid product or solid product is become to powder productSuddenly.
The present invention also provides the application of schizophyllum commune in the preparation fermentate of resistance to anoxic.
The present invention has found the new application in the preparation fermentate of resistance to anoxic of schizophyllum commune this bacterial classification, with this bacterial classification for setting outBacterial classification is produced the culture medium of the fermentate of resistance to anoxic, and composition is simple, and raw material sources are extensive, and cost is low, is easy to preparation; MovingThing experiment showed, that the fermentate of resistance to anoxic of the present invention has with the product of resistance to anoxic confirming the validity and has identical resistance to lackingOxygen effect; The fermentate of resistance to anoxic of the present invention, after continuously edible certain hour, can improve the tolerance of body to hypoxemiaPower, is conducive to alleviate and alleviate a series of anoxic symptoms such as altitude sickness; The present invention produces the method for the fermentate of resistance to anoxicTechnique is simple, low to production equipment requirement, saves the energy, and cost is low, is easy to large-scale industrial production, has wideWealthy prospects for commercial application.
Brief description of the drawings
Fig. 1 is schizophyllum commune (Schizophyllum commune) shaking flask cultivation and fermentation conditional curve
Fig. 2 is schizophyllum commune (Schizophyllum commune) fermentation tank culture sweat curve
Fig. 3 is that schizophyllum commune (Schizophyllum commune) is 1#Static culture sweat curve in plate
Fig. 4 is that schizophyllum commune (Schizophyllum commune) is 2#Static culture sweat curve in plate
Fig. 5 A-5G is that schizophyllum commune (Schizophyllum commune) is 1#In plate, the growing state of 1-7 days shinesSheet
Fig. 6 A-6G is that schizophyllum commune (Schizophyllum commune) is 2#In plate, the growing state of 1-7 days shinesSheet
Detailed description of the invention
In the present invention, comprise in highland barley, barley, wheat and maize etc. as the cereal section crop of culture medium primary raw materialAny, refer to the powder of its seed body, seed body, the powder juice of seed body, the sprout of seed body, the bud of seed bodyThe bud juice of powder and seed body. Wherein highland barley, from Qinghai-Tibet Platean, refers to highland barley seed body, highland barley flour (highland barley seedBody grinds), highland barley flour juice, highland barley bud, highland barley bud powder or highland barley bud juice.
Nitrogenous source refers to peptone, yeast extract, ammonium chloride, sulfuric acid amine or ammonium nitrate.
Schizophyllum commune (Schizophyllum commune) can be AS 5.53, AS 5.113, and AS 5.115, AS 5.116,AS 5.117, AS 5.118, AS 5.119, AS 5.120, AS 5.131 and AS 5.386 (Chinese Academy of Sciences microorganismCulture presevation DSMZ) in any one or its any combination.
The preparation of embodiment 1, schizophyllum commune culture medium
1, the preparation of highland barley bud, highland barley bud powder, highland barley bud juice
Highland barley seed body is removed to excessive moisture after 17 DEG C of immersion 4h, make it keep moistening 20h, 17 DEG C of germinationsAfter 2 days, obtain highland barley bud. Highland barley bud, 60 DEG C of oven dry, is removed to root, pulverized after 40 mesh sieves, obtain green grass or young cropsHighland barley bud powder. Highland barley bud powder adds 60 DEG C of about 4h of insulation of water (every 0.5h stirs once) of 4 times of volumes, with the inspection of iodine liquidObtain highland barley bud juice without starch, every 1 part of highland barley seed body is produced 3.75 parts of highland barley bud juice.
2, the preparation of highland barley flour, highland barley flour juice
Highland barley seed body was pulverized to 40 mesh sieves and obtained highland barley flour.
200g highland barley flour is added to 1L water boil 30min, and four layers of filtered through gauze, get juice, are settled to 1L and obtain highland barley flourJuice, for preparing culture medium.
While adopting other cereal section crop, prepare corresponding culture medium by identical method.
3, schizophyllum commune culture medium
(1) slant medium: PDA culture medium (g/L): potato 200.0, glucose 20.0, MgSO4·7H201.5，
KH2PO43.0, Cobastab10.01, agar 20, adds 20% murphy juice to 1L;
(2) first order seed culture medium (g/L): glucose 30.0, analysis for soybean powder 10.0, yeast extract 1.0, MgSO4·7H2O
0.5，KH2PO41.0, Cobastab10.01，pH6.0；
Secondary seed medium: 12 ° of Brix highland barley bud juice, 0.5%NH4Cl；
(3) fermentation medium
There is following several fermentation medium:
Fermentation medium 1:12 ° of Brix highland barley bud juice, 0.5%NH4HCl。
Fermentation medium 2: highland barley flour juice, 0.1% peptone.
Fermentation medium 3: highland barley flour, 0.5%NH4SO4, 1 times to the water of highland barley flour quality.
Fermentation medium 4: wheat flour, 0.5%NH4NO3, 2 times to the water of flour quality.
Fermentation medium 5:10 ° of Brix fructus hordei germinatus juice, 0.5%NH4Cl. Wherein, 10 ° of Brix fructus hordei germinatus juice sides routinelyLegal system is standby.
Fermentation medium 6: corn flour, 0.5% yeast extract, 3 times are to the water of corn flour quality.
Fermentation medium 7: highland barley bud powder, 0.5%NH4SO4, 1 times to the water of highland barley bud opaque amount.
Concentration described here (%) refers to the mass percent of solute (as nitrogenous source) in culture medium solution.
Brix is the unit of soluble solid in highland barley bud juice (mainly referring to sugar) content.
Above-mentioned three class medium pH natures.
Embodiment 2, liquid culture under aeration are prepared the fermentate of resistance to anoxic
The production technology that liquid culture under aeration is prepared the fermentate of resistance to anoxic specifically can comprise the following steps:
(1) slant strains is cultivated
Schizophyllum commune (Schizophyllum commune) AS 5.53 use inoculation hooks are cut to 1cm from inclined-plane2Band culture mediumInclined-plane mycelia proceed to slant medium in embodiment 1 middle part, 26 DEG C cultivate 6d (my god).
(2) level liquid seed culture
From step (1), in cultured inclined-plane, cut 1cm with inoculating hook2Inclined-plane mycelia access with culture medium is equipped with oneIn the 500mL triangular flask (liquid amount is 30%) of level seed culture medium, at 26 DEG C, 150rpm, cultivation 3D (my god)Obtain liquid seeds.
(3) secondary liquid seeds is cultivated
Cultured level liquid seed is equipped with to the fermentation tank (dress of secondary seed medium according to 10% inoculum concentration accessLiquid measure is 70%) in, 26 DEG C, 150rpm, cultivates 3D.
(4) fermentation tank culture
In 7L fermentation tank, pack 5L fermentation medium 1 into. By cultured liquid seeds in step (3) according to culture mediumIn the inoculum concentration access fermentation tank of charge weight 10% (0.5L), dissolved oxygen control is 30%, at 26 DEG C, and speed of agitator200rpm, throughput is 150L/h, fermentation 5D.
(5) extract the fermentate of resistance to anoxic
Stop, after fermentation, taking out zymotic fluid, process after filtration, by thalline and separation of fermentative broth, remove thalline, filtrateBe the fermentate of resistance to anoxic.
(6) post processing of the fermentate of resistance to anoxic
1. the concentrated fermentate of resistance to anoxic of Vacuum film evaporation device, controls temperature below 70 DEG C, treats the dry of zymotic fluidConcentration reaches 50% termination concentration.
2. batching, granulation
By the raw material such as zymotic fluid and converted starch, natural sweetener, acid and natural essence after concentrated at batch mixerMiddle wet-mixing, mixed material passes through granulator granulation.
3. dry
The granule materials of making is passed through to fluidized bed drying.
Embodiment 3, liquid static culture legal system are for the fermentate of resistance to anoxic
Its production technology specifically can comprise the following steps:
(1) slant strains cultivation, (2) level liquid seed culture and (3) secondary liquid seeds are cultivated, methodWith embodiment 2;
(4) static fermentation is cultivated
Cultured liquid seeds in step (3) is accessed in culture vessel according to the inoculum concentration of 10% (volume ratio),
1m2Culture vessel dress 1.94L fermentation medium 2 or fermentation medium 5, media surface area 3.44 (m2/m3),26 DEG C of static cultivation 7D (my god).
(5) extract the fermentate of resistance to anoxic
Stop, after fermentation, taking out zymotic fluid, after filtration by thalline and separation of fermentative broth, remove after thalline, filtrate isThe fermentate of resistance to anoxic.
(6) post processing of the fermentate of resistance to anoxic, identical with embodiment 2.
Embodiment 4, solid medium cultivation are prepared the fermentate of resistance to anoxic
Specifically comprise the following steps:
(1) slant strains cultivation, (2) level liquid seed culture and secondary liquid seeds are cultivated with embodiment 2,Wherein selecting bacterial strain is AS 5.386;
(3) solid state fermentation is cultivated
By cultured liquid seeds in step (2) according to 10% (seed volume ml/ dry matter content inoculation g)Amount evenly point is received the solid medium in culture vessel, and container dress 1856g fermentation medium 3,4 or 6 or 7 exists30 DEG C cultivate 7D (my god), the solid-state culture (comprising culture medium and mycelium) obtaining is the fermentate of resistance to anoxic.
(4) post processing of the fermentate of resistance to anoxic
1. dry
Stop, after fermentation, the fermentate of resistance to anoxic being taken out, 60 DEG C of oven dry.
2. powder process
The product of drying is pulverized, sieved and obtain powder product.
Embodiment 5, schizophyllum commune (Schizophyllum commune) sweat analysis
1,500mL triangular flask shaking flask is cultivated
Cultured secondary liquid seeds in embodiment 2 is equipped with to fermentation training according to 10% inoculum concentration (volume ratio) accessSupport in the 500mL triangular flask (liquid amount is 10%) of base 1,26 DEG C, 150rpm, gets sample one time every 12h, surveysDetermine total sugar content, polyoses content, content of reducing sugar, amino nitrogen content, thalline biomass and pH.
Its sweat curve as shown in Figure 1, shows:
In whole sweat, pH remains on 5.0 left and right substantially, and the later stage decreases a little, according to existing reportAnalyze, this is because schizophyllum commune can produce a small amount of malic acid during the fermentation.
The variation more complicated of amino nitrogen content. 0-24h declines, and 24-36h rises to some extent, from 36h to fermentation terminationAmino nitrogen content is along with the growth of thalline declines gradually. This is due at earlier fermentation, and thalline utilizes in zymotic fluid easierThe ammonia nitrogen utilizing, therefore ammonia nitrogen slightly declines; Along with the growth of thalline, mycelia can secrete a small amount of protease,Thereby the proteins and peptides in decomposing and fermenting liquid produces amino acid, thereby amino nitrogen content rises; To mid-term mycelia fastSpeed increases, and amino nitrogen content is also consumed fast, to synthesize thalline; Along with the consumption of nutriment, thalline starts certainlyMolten, discharge a small amount of ammonia nitrogen.
Changes of Reducing Sugar Content general trend is to reduce gradually with incubation time, and the speed of decline is slower. Total sugar contentVariation tendency is similar to reduced sugar.
Polyoses content reaches to the highest at 4D, is 0.82g/L.
2,7L fermentation tank batch culture
Cultured secondary liquid seeds in embodiment 2 is equipped with to fermentation according to 10% inoculum concentration (volume ratio) accessIn the 7L fermentation tank (liquid amount 75%) of culture medium 1, dissolved oxygen control is 30%, at 26 DEG C, and speed of agitator 200rpm,Throughput is 150L/h, gets sample one time every 12h, measures total sugar content, polyoses content, content of reducing sugar, aminoNitrogen content, thalline biomass and pH.
The change curve of its sweat as shown in Figure 2, shows:
The final pH of fermentation tank is 4.2 left and right, compared with the final pH of shaking flask low (final pH of shaking flask is 4.8 left and right).Content of reducing sugar, variation tendency and the shaking flask of total sugar content are basically identical. Amino nitrogen content is wanting than shaking flask on the wholeHigh. Thalline biomass, polyoses content reach to the highest at 4d, and thalline biomass is 18.45g/L, and polyoses content is1.182g/L, polyoses content is cultivated and will be exceeded 0.362g/L than shaking flask.
3, static liquid cultivation
Cultured secondary liquid seeds in embodiment 3 is equipped with to fermentation according to the inoculum concentration access of 10% (volume ratio)1 of culture medium 2#(diameter is 155mm) or 2#(diameter is 128mm), in 26 DEG C, static culture 7d, every 24hGet sample one time, measure total sugar content, polyoses content, content of reducing sugar, amino nitrogen content, thalline biomass and pH.
1#The plate static fermentation thalline growing state of 1-7 days, as shown in Fig. 5 A-5G, shows the 1st day, cultivates base tableFace has had fragmentary white hypha bead; 2-3 days, fragmentary white hypha ball is grown up gradually; 4-6 days,The area of mycelium pellet and highly further increase, form large stretch of mycelium gradually; Nearly cover while termination to fermentationThe surface of whole zymotic fluid.
2#The plate static fermentation thalline growing state of 1-7 days as shown in Fig. 6 A-6G, show its growing state basic with
1#Plate is identical.
As shown in Figures 3 and 4, Fig. 3 shows 1 to the result of variations of its sweat#The dynamic fermentation of plate and air blow and agitationCompare: content of reducing sugar, the trend that total sugar content reduces is slower than dynamic agitation aerlbic culture; PH, amino nitrogen contentChange curve is substantially similar; When thalline biomass reached at the 5th day when the highest, polyoses content also reaches to the highest, and this saysBright exocellular polysaccharide is accompanied by thalli growth and produces; Polyoses content reaches the highest at 5d. Fig. 4 shows 2#PlateThalline biomass reaches to maximum at 5D. Polyoses content reaches to maximum at 6D, than 1#Polysaccharide yield improves. This saysBright in the time that other condition is identical, select 2#Condition of culture is than 1#More be conducive to the generation of polysaccharide.
Static culture is compared with fermentation tank culture with shaking flask: static culture polyoses content is the highest, be respectively shaking flask cultivate and4.01 times and 2.78 times of fermentation tank culture; Static culture obtains maximum thalline biomass at 5d, is 19.00g/L
(2#Plate) and 25.20g/L (1#Plate); Shaking flask is cultivated and is obtained maximum thalline biomass at 4D, for15.100g/L; Fermentation tank culture also obtains maximum thalline biomass at 4d, is 18.448g/L; Static cultureTime is longer, and 1#Plate reaches to maximum at 5D polyoses content, is 2.050g/L; 2#Plate contains at 6D polysaccharideAmount reaches to maximum, is 3.290g/L; Shaking flask is cultivated the schizophyllan content obtaining and is reached to maximum at 4D,For 0.820g/L; Fermentation tank culture also reaches to maximum at 4D, is 1.182g/L.
Embodiment 6, zoopery
Airtight anoxia zoopery material
250mL ground wide-mouth bottle, syringe (syringe needle is oblate, can avoid making animal in gavage process injured),Stopwatch, vaseline, soda lime (or equivalent NaOH and calcium carbonate, the moisture absorption); (Chinese Academy of Sciences is moving for small white mouseThing institute), male, body weight 20 ± 2g, 20 ± 5 DEG C of cultivation temperature.
Every group of 12 mouse, every mouse feeds the 0.5ml sample (dry matter content in 2#, 3#, 5# sample at every turnBe 50%; ), feed 1 every day. Take syringe gavage mode to feed, 21 days nursing time. For the last timeAfter feeding 30min, start to test mouse hypoxia-bearing capability. Mouse is put into the wide-mouth bottle that fills in advance 15g soda lime,Be coated with after vaseline at bottleneck, cover bottle cap, start timing, write down the time-to-live of mouse in airtight wide-mouth bottle.Wherein, the wide-mouth Bottle and bottle cap size homogeneous of selecting, to ensure that in wide-mouth bottle, the content of oxygen is consistent. Wide-mouth bottle is airtightObturage, be strictly on guard against gas leakage.
Water, 981 dietary supplements, Hericium erinaceus attenuate liquid, schizophyllum commune solid state fermentation thing (adopt highland barley bud powder,Embodiment 4 makes) and schizophyllum commune highland barley bud juice zymotic fluid (adopt highland barley bud juice, embodiment 2 makes) lack mouse is airtightThe impact of oxygen time-to-live is as shown in table 1:
Table 1.
* for significant difference, * * are difference highly significant
1#, 2# are control sample (the negative control sample of 1#; Positive control sample-981 of 2# sample dietary supplements, is derived from forceChinese defense economy institute. It is that the various plants such as a kind of employing ginseng, matrimony vine, rhizoma Gastrodiae are raw material, makes by lixiviateThe plant extracts with anti-anoxic effect, Zeng Huo army scientific and technological progress second prize. ).
Table 1 shows that 981 dietary supplements and water ratio have significant difference (T0.01＞T＞T0.05), illustrate that this fermentate hasSignificant anoxia tolerance function; Highland barley bud juice schizophyllum commune fermentate (prepared by embodiment 2) has very aobvious with water ratioWork difference (T > T0.01), illustrate that this fermentate has the anoxia tolerance function of highly significant; Schizophyllum commune solid state fermentation thing is (realExecute prepared by example 4) there is highly significant difference (T > T with water ratio0.01), it is poor not have compared with 981 dietary supplementsDifferent (T < T0.05), illustrate that this fermentate has the anoxia tolerance function of highly significant; Hericium erinaceus zymotic fluid compares all not tool with waterThere were significant differences (T < T0.05), illustrate that Hericium erinaceus zymotic fluid does not have anoxia tolerance function.
Embodiment 7, zoopery
Every group of 12 mouse, every mouse feeds the 0.5ml sample (dry matter content in 2#, 3# and 4# sample at every turnBe 25%; 4# is zymotic fluid prepared by the embodiment 3 schizophyllum commune tunning when being concentrated into dry matter content and being 25%),Feed 1 every day. Take syringe gavage mode to feed, 21 days nursing time, the equal well-grown of all mouse, notThere is death and side effect. After last feeding 30min, start to test mouse hypoxia-bearing capability. Mouse is put in advanceFirst fill in the wide-mouth bottle of 15g soda lime, be coated with after vaseline at bottleneck, cover bottle cap, start timing, write down mouseTime-to-live in airtight wide-mouth bottle. Wherein, the wide-mouth Bottle and bottle cap size homogeneous of selecting, to ensure wide-mouth bottleThe content of middle oxygen is consistent. Wide-mouth bottle is airtight to be obturaged, and is strictly on guard against gas leakage.
Water, 981 dietary supplements, Fermentation Condition of Monascus spp thing and schizophyllum commune fructus hordei germinatus juice zymotic fluid are to the mouse survival timeAffect as table 2:
Table 2: water, 981 dietary supplements, Fermentation Condition of Monascus spp thingAnd the impact of fructus hordei germinatus juice schizophyllary fermentating liquid on the mouse airtight anoxia time-to-live
Note: 1#, 2# are control sample (the negative control sample of 1#; Positive control sample-981 of 2# sample dietary supplements, sourceFrom Wuhan defense economy institute. It is that the various plants such as a kind of employing ginseng, matrimony vine, rhizoma Gastrodiae are raw material, by lixiviateThe plant extracts with anti-anoxic effect of making, Zeng Huo army scientific and technological progress second prize. );
* for significant difference, * * are difference highly significant
Table 2 shows that 981 dietary supplements and water ratio have highly significant difference (T > T0.01), 981 meals benefits are describedFill the anoxia tolerance function that agent also has highly significant; Fructus hordei germinatus juice schizophyllum commune fermentate (zymotic fluid prepared by embodiment 3Schizophyllum commune tunning when being concentrated into dry matter content and being 25%) there is highly significant difference (T > T with water ratio0.01)，Do not there is difference (T < T compared with 981 dietary supplements0.05), illustrate that this fermentate has the anoxia tolerance function of highly significant;Fermentation Condition of Monascus spp thing with water than not thering is highly significant difference (T < T0.03), illustrate that Fermentation Condition of Monascus spp thing does not have resistance to lackingOxygen function.
With method is identical, use highland barley flour, little to using in embodiment 3 in the fermentation of highland barley flour juice, embodiment 4 aboveThe tunning that flour, fermentation of corn starch obtain is fed the mouse survival time and is measured, and has verified equally these fermentationsProduct has anoxia tolerance function.
Claims (16)
1. produce the fermentate of a resistance to anoxic culture medium taking schizophyllum commune as starting strain, with highland barley, barley, wheat andIn corn cereal section crop, the sprout of any seed body, the bud powder of seed body or the bud juice of seed body are for mainly formerMaterial.
2. culture medium according to claim 1, is characterized in that: described culture medium also comprises nitrogenous source, nitrogenous sourceFor one or more the combination in peptone, yeast extract, ammonium chloride, sulfuric acid amine and ammonium nitrate.
3. culture medium according to claim 1 and 2, is characterized in that: described cereal section crop is highland barley.
4. culture medium according to claim 3, is characterized in that: described culture medium is 10-14 ° of Brix highland barleyBud juice and 0-0.5%NH4Cl。
5. produce a method for the fermentate of resistance to anoxic, comprise taking schizophyllum commune (Schizophyllum commune) asStarting strain, fermentation claim 1 to 4 arbitrary described culture medium, obtains the step of the fermentate of resistance to anoxic; Bag successivelyDraw together the process of slant strains cultivation, level liquid seed culture, the cultivation of secondary liquid seeds and fermented and cultured, wherein:Secondary liquid seed culture medium is by 12 ° of Brix highland barley bud juice and 0.5%NH4Cl composition; Described fermented and cultured is liquid logicalGas fermentation, culture medium is 10-14 ° of Brix highland barley bud juice and 0-0.5%NH4Cl, condition of culture is fermentation temperature 15-35DEG C, air blow and agitation when fermentation, fermentation time is 3-6D.
6. the method for producing according to claim 5 the fermentate of resistance to anoxic, is characterized in that: also comprise fermentationThing post processing forms the step of solid product.
7. the method for producing according to claim 5 the fermentate of resistance to anoxic, is characterized in that: also comprise fermentationThing post processing forms the step of powder product.
8. produce a method for the fermentate of resistance to anoxic, comprise taking schizophyllum commune (Schizophyllum commune) asStarting strain, fermentation claim 1 to 4 arbitrary described culture medium, obtains the step of the fermentate of resistance to anoxic; Bag successivelyDraw together the process of slant strains cultivation, level liquid seed culture, the cultivation of secondary liquid seeds and fermented and cultured, wherein:Secondary liquid seed culture medium is by 12 ° of Brix highland barley bud juice and 0.5%NH4Cl composition; Described fermented and cultured is liquid quietState fermentation, described fermentation medium is 10-14 ° of Brix highland barley bud juice and 0-0.5%NH4Cl; 26 DEG C of described fermentation temperatures,Described fermentation time 4D.
9. the method for producing according to claim 8 the fermentate of resistance to anoxic, is characterized in that: also comprise fermentationThing post processing forms the step of solid product.
10. the method for producing according to claim 8 the fermentate of resistance to anoxic, is characterized in that: also comprise fermentationThing post processing forms the step of powder product.
Produce the method for the fermentate of resistance to anoxic for 11. 1 kinds, comprise with schizophyllum commune (Schizophyllum commune)For starting strain, fermentation claim 1 to 4 arbitrary described culture medium, obtains the step of the fermentate of resistance to anoxic; SuccessivelyComprise that slant strains cultivation, level liquid seed culture, secondary liquid seeds are cultivated and the process of fermented and cultured, wherein:Secondary liquid seed culture medium is by 12 ° of Brix highland barley bud juice and 0.5%NH4Cl composition; Described fermented and cultured is solid-state quietState fermentation, culture medium is by highland barley bud powder, 0.5% (NH4)2SO4, 1 times of composition of the water to highland barley bud opaque amount, cultivateCondition is fermentation temperature 15-35 DEG C, when fermentation, leaves standstill and cultivates, and fermentation time is 3-6D.
12. according to the method for producing the fermentate of resistance to anoxic described in claim 11, it is characterized in that: also comprise and will send outThe post processing of ferment thing forms the step of powder product.
13. 1 kinds of fermentates of resistance to anoxic are tunnings that method obtains described in claim 5 or 8; Described fermentationProduct is in zymotic fluid, to remove the liquid product after thalline, wherein content of reducing sugar 15-40g/L, total sugar content30-70g/L, thalline biomass 10-30g/L.
14. 1 kinds of fermentates of resistance to anoxic are solid products that method obtains described in claim 6 or 9.
15. 1 kinds of fermentates of resistance to anoxic are solid products that method obtains described in claim 11.
16. 1 kinds of fermentates of resistance to anoxic are powders that method obtains described in claim 7 or 10 or 12.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828664A (en) * | 2017-10-25 | 2018-03-23 | 湖南省农业生物技术研究中心 | Schizophyllum commune XT 1 and its cultural method and application |
| CN109220524A (en) * | 2018-09-29 | 2019-01-18 | 云南菌视界生物科技有限公司 | A kind of super short period cultural method of schizophyllum commune bacterial strain and its fructification |
-
2004
- 2004-03-15 CN CN200410028929.1A patent/CN106342092B/en not_active Expired - Lifetime
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828664A (en) * | 2017-10-25 | 2018-03-23 | 湖南省农业生物技术研究中心 | Schizophyllum commune XT 1 and its cultural method and application |
| CN107828664B (en) * | 2017-10-25 | 2020-10-23 | 湖南省农业生物技术研究中心 | Schizophyllum commune XT-1 and cultivation method and application thereof |
| CN109220524A (en) * | 2018-09-29 | 2019-01-18 | 云南菌视界生物科技有限公司 | A kind of super short period cultural method of schizophyllum commune bacterial strain and its fructification |
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