CN107267398A - A kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum cultural method - Google Patents

A kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum cultural method Download PDF

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Publication number
CN107267398A
CN107267398A CN201710454702.0A CN201710454702A CN107267398A CN 107267398 A CN107267398 A CN 107267398A CN 201710454702 A CN201710454702 A CN 201710454702A CN 107267398 A CN107267398 A CN 107267398A
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ganoderma lucidum
liquid strain
liquid
culture medium
strain
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CN107267398B (en
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张国民
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National Breeding Farm Of Jiangyin Mayor Jing River
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National Breeding Farm Of Jiangyin Mayor Jing River
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor

Abstract

The invention discloses a kind of Liquid Strain of Ganoderma Lucidum culture medium prescription, including：Peptone 6g, glucose 15g, magnesium sulfate 0.5g, potassium dihydrogen phosphate 0.5g, farina 1g, corn, tapioca 3g, vitamin B110mg, agar 1g, pure water 1000ml.And Liquid Strain of Ganoderma Lucidum cultural method：Based on Liquid Strain of Ganoderma Lucidum culture medium prescription, through high-temperature sterilization, it is inoculated with into parent species, then through shaking table culture, obtains Liquid Strain of Ganoderma Lucidum.Such a Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum reduce incubation step, shorten mycelium growing period, shorten that growth cycle, contamination rate be low, cost is low, and yield is high, it is adaptable to scale, factorial praluction.

Description

A kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum cultural method
Technical field
The present invention relates to fungus growing technique field, more particularly to a kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and ganoderma lucidum Strain cultivation method.
Background technology
The training method of current lucidum strain is mainly on this traditional planting type of solid culture, using traditional cultivation The ganoderma lucidum that training mode is produced has been difficult to the demand for meeting domestic and international market in terms of quality and yield.Solid spawn productive culture Ganoderma lucidum have that bacterium germination is slow and cell age is inconsistent, premunition is weak, and vitality is not strong, is vulnerable to the shortcomings of miscellaneous bacteria infects.
The content of the invention
The present invention need to provide a kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum cultural method, solve solid The ganoderma lucidum of strain productive culture has that bacterium germination is slow and cell age is inconsistent, and premunition is weak, and vitality is not strong, is vulnerable to miscellaneous bacteria infection etc. Technical problem.
In order to solve the above technical problems, the present invention provides a kind of Liquid Strain of Ganoderma Lucidum culture medium prescription, including：Peptone 6g, glucose 15g, magnesium sulfate 0.5g, potassium dihydrogen phosphate 0.5g, farina 1g, corn, tapioca 3g, vitamin B110mg, agar 1g, pure water 1000ml.
The present invention provides another Liquid Strain of Ganoderma Lucidum culture medium prescription, peptone 250g, glucose 1000g, magnesium sulfate 40g, potassium dihydrogen phosphate 20g, farina 100g, corn, tapioca 200g, vitamin B1600mg, agar 100g, disappear Infusion 5g, pure water 60L.
The present invention also provides a kind of Liquid Strain of Ganoderma Lucidum cultural method, comprises the following steps：
(1) prepare the triangular flask of multiple 500ml capacity, 200ml/ bottles are respectively charged into multiple triangular flasks by claim Multiple triangular flasks are placed on high steam pot sterilizing by culture medium made from the Liquid Strain of Ganoderma Lucidum culture medium prescription described in 1 or 2, Then take out cooling；
(2) ganoderma lucidum slant tube parent species one are taken, 1 piece of picking broad bean size or 2 ferfas kinds are accessed in each triangular flask, so It is quiet to 24 hours afterwards；
After (3) 24 hours, multiple triangular flasks are put into rotary shaker, cultivated on shaking table, shaking table culture 6 days, are made Vial of liquid strain.
It is high described in step (1) as a kind of a kind of preferred scheme of Liquid Strain of Ganoderma Lucidum cultural method of the present invention It is specially high-temperature sterilization 30 minutes under the conditions of 120 DEG C -126 DEG C to press steam copper sterilizing.
As a kind of a kind of preferred scheme of Liquid Strain of Ganoderma Lucidum cultural method of the present invention, step is shaken described in (3) The rotary speed of bed is in 120-130r/min, and temperature is 25 DEG C-30 DEG C.
As a kind of a kind of preferred scheme of Liquid Strain of Ganoderma Lucidum cultural method of the present invention, step also includes after (3) Following steps：
(4) prepare the triangular flask of Large Copacity, vial of liquid strain made from step (3) is accessed into big triangular flask, loading amount 4 Liter/bottle liquid spawn culture medium, rotary shaker is put into by the triangular flask of the Large Copacity, is cultivated 36 hours on shaking table；
(5) liter/bottle liquid spawn of loading amount 4 is accessed into 60 liters of stainless steel air blow tank, in the air blow tank Culture 36 hours, is made Liquid Strain of Ganoderma Lucidum；
(6) by the Liquid Strain of Ganoderma Lucidum in fermentation tank, the bag for accessing ganoderma lucidum solid cultivating in bag strain is injected with syringe Oral area position, carries out large-scale production.
As a kind of a kind of preferred scheme of Liquid Strain of Ganoderma Lucidum cultural method of the present invention, in step (5), supply is logical The air of gas fermentation tank is to pass through the filtrated air after a series of purified treatments of air sterilization and filtering system.
The beneficial effect of a kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum cultural method of the present invention Fruit is：Liquid Strain of Ganoderma Lucidum efficiently, conveniently, safe and pollution-free, lucidum strain steady quality, with short production cycle, bacterium germination is fast, Production is not influenceed by season, goes out sesame soon, premunition is strong, yield is high, can carry out scale, factorial praluction.According to ganoderma lucidum Optimal growth conditions, using optimal environmental Kuznets Curves pattern, batch production carries out extensive high-volume cultivation production, make ganoderma lucidum not by Season and the influence of environment, realize annual controllable, stable efficient productive target.
Embodiment
In order to facilitate the understanding of the purposes, features and advantages of the present invention, with reference to embodiment The present invention is further detailed explanation.
Embodiment 1
Make 1000ml liquid spawns formula：Peptone 6g, glucose 15g, magnesium sulfate 0.5g, potassium dihydrogen phosphate 0.5g, Farina 1g, corn, tapioca 3g, vitamin B11 (10 milligrams), agar 1g, pure water 1000ml.
Vial of liquid Spawn incubation method：
Prepare the triangular flask of 500ml capacity, triangular flask is distributed into 200ml/ bottles of liquid spawn culture mediums, triangular flask is put Sterilized in high steam pot, in 120 DEG C -126 DEG C high-temperature sterilizations 30 minutes, then take out cooling, take ganoderma lucidum slant tube parent species One, 1 piece or 2 pieces of picking broad bean size is accessed in triangular flask, quiet to 24 hours.After 24 hours, triangular flask is put into rotatably Shaking table, is cultivated on shaking table, and the characteristic of shaking table can influence the matter of oxygen to pass effect, and then influence mycelial growth, therefore control Rotary speed, control is in 120-130r/min, and temperature is 25 DEG C-30 DEG C optimums, shaking table culture 6 days.
Embodiment 2
Make 60 liters of liquid spawn formulas：Peptone 250g, glucose 1000g, magnesium sulfate 40g, potassium dihydrogen phosphate 20g, Farina 100g, corn, tapioca 200g, vitamin B160, agar 100g, 1 spoonful of defoamer (5 grams), pure water 60L。
Access the cultural method of fermentation tank：
1. preparing the triangular flask of 500ml capacity, triangular flask is distributed into 200ml/ bottles of liquid spawn culture mediums, by triangular flask High steam pot sterilizing is placed on, in 120 DEG C -126 DEG C high-temperature sterilizations 30 minutes, cooling is then taken out, takes ganoderma lucidum slant tube mother Plant in one, 1 piece or 2 pieces access triangular flask of picking broad bean size, it is quiet to 24 hours.After 24 hours, triangular flask is put into rotation Formula shaking table, is cultivated on shaking table, and the characteristic of shaking table can influence the matter of oxygen to pass effect, and then influence mycelial growth, therefore control Rotational speed processed, control is in 120-130r/min, and temperature is 25 DEG C-30 DEG C optimums, shaking table culture 6 days.
2. preparing the triangular flask of Large Copacity, vial of liquid strain is accessed into big triangular flask, the training of liter/bottle liquid spawn of loading amount 4 Base is supported, triangular flask is put into rotary shaker, is cultivated 36 hours on shaking table.
3. liter/bottle the liquid spawn of loading amount 4 to be accessed to 60 liters of stainless steel fermentation tank, 36 are cultivated in air blow tank small When, in whole incubation, it is desirable to which the air of supply is sterile, by a series of purified treatments of air sterilization and filtering system, Air cleaning enters in fermentation tank into sterile air.Effect of the filtrated air in fermented and cultured is to provide oxygen, promotes training Nutrient solution is stirred, and mycelium is constantly mixed in the medium, improve to absorption of nutrient ingredients with effect, maintenance metabolism product it is uniform Diffusion, prevents microbiological contamination in fermentation process.In incubation, observation temperature, pressure, mixing speed, gas and fluid flow, liquid Body viscosity, turbidity, ventilation exhaust, mythic fungus bacterium ball state, note the outward appearance of Ganoderma lucidum by submerged culture liquid, such as clarification, muddy, wadding at any time Shape etc., while be also noted that color, smell etc., as Liquid Strain of Ganoderma Lucidum has special fragrant, mould has vinosity, bacterium acid Taste, peculiar smell or tasteless.
4. by the Liquid Strain of Ganoderma Lucidum in fermentation tank, the sack for accessing ganoderma lucidum solid cultivating in bag strain is injected with syringe Position, carries out large-scale production.
In the above-described embodiments, grown in the mycelia liquid medium within of ganoderma lucidum, absorb in liquid medium within moisture and Nutrition, turns out mycelium, and mycelium is secondary mycelia, and it is more sturdy than the mycelia that spore germination is formed, fast growth. The mycelia that observation Liquid Culture goes out in appearance has two kinds of forms：One kind is common filamentous, and another is orbicule.Due to bacterium Silk growth be to be extended radially to surrounding, in liquid medium within ventilation or stirring action under, mycelia is in original culture It is grown into base, or grows branch and crisscross, mutual winding and subsphaeroidal, referred to as mycelium pellet.Mycelium pellet form of diverse, It is general radial, with jagged, in 4-8MM or so more than diameter.Some bacterium ball surfaces are smooth, and some rough surface have seemingly Seta ball.Culture medium (liquid) be mycelia depend on for existence, the place of mass exchange and metabolite, the composition of culture medium is for mycelia Growth and breeding, metabolite synthesis all have important influence, be related to the yield and quality of fermented product.Want to obtain Good ganoderma lucidum harvest, the breeding of its mycelium pellet is still important, and every forming a large amount of mycelium pellets, its nutrient solution is more clear, conversely not Form mycelium pellet or a small amount of forming mycelium pellet, be then that nutrient solution is more muddy.It follows that nutrient solution is muddy, more not Be conducive to the growth of mycelia.
The mycelium pellet form of ganoderma lucidum is also influenceed by medium component and PH acid-base values and its condition of culture：
First, culture medium contains starch component, and its viscosity is bigger, and bacterium ball is smaller, and seta is shorter, and Ganoderma lucidum mycelium is in Ma Ling Most fast, other then poor, although in existing technology, some sections are grown in the soluble starches such as sweet potato starch, corn tapioca Unit is ground to try to be used as the source of culture medium starch, still, its training being made by the use of the crop products such as potato and corn Foster base is very muddy, and medium component is uneven, hinders the growth of mycelium pellet, and culture medium is also easily denatured.
Secondly, under different PH, mycelium can be thread or spherical；Culture medium prescription in this programme PH value in 5.0-6.0, the PH scopes such as phosphate concn in the medium for being the growth of Ganoderma lucidum mycelium optimum is 0.1%, raw Length is slower, and bacterium ball is big, if concentration is reduced, bacterium ball is just small.
Furthermore, inoculum concentration and throughput size in incubation, mixing speed speed are also produced to thalli morphology change Influence.When inoculum concentration is big, it can generally be grown in decentralized, and inoculum concentration hour, then be conducive to the formation of bacterium ball.Generally it is inoculated with Amount is bigger, and bacterium bulb diameter is just smaller, and inoculum concentration is smaller, and bacterium bulb diameter is bigger.During general shaking table use, utilization The method of vibration, and it has the disadvantage to spin upside down, reach above and below it is uniform, but do not reach the uniform of same level, so, In the technical program, shaking table is made one and improved, an agitating shaft is added, the agitation of level is carried out to it, mixing speed is fast or ventilates When measuring big, bacterium bulb diameter is just small, and mixing speed is slow or throughput hour, and bacterium bulb diameter is just big.So, the mixing speed control of shaking table System, in 120-130r/min rotating speeds, is bacterium solution so on shaking table liquid dynamic cultivation upper strata, lower floor is mycelium pellet, is more beneficial for The growth of strain.
In addition, when concentration sugared in culture medium is generally sugar excess in 2%, culture medium, the dissolving supported in air is often hindered, Be unfavorable for mycelial growth of sinking, therefore, in the technical program, control emphatically sugar concentration, some existing laboratories, The sugared part content made is higher than 2%, and the experiment culture that can only really apply under laboratory condition is unsuitable for the agriculture of mass production Sparetime university's large-scale production, also without practical operation prospect.
Further, the growth of Ganoderma lucidum mycelium needs the effect of some micro growth factors, and growth factor is one kind in mycelia Itself can not be synthesized in growth course, it is necessary to the material of the beneficial mycelia growth and breeding of a class obtained from the external world, in practice it has proved that, Vitamin B1 is added in Ganoderma lucidum by submerged culture base can remarkably promote the growth of mycelia.
Finally, contain protein substance in Ganoderma lucidum by submerged culture base, foam is easily formed under ventilation and stirring, training is caused Liquid rising is educated, if not being controlled by that the substantial amounts of chance escaped liquid, add microbiological contamination can be caused.Simultaneously as foam is excessive, can also What influence was ventilated and stirred is normally carried out, so as to influence the successful incubation of Liquid Strain of Ganoderma Lucidum, therefore, only add froth breaking Agent, just can guarantee that cultivation.
With the plantation of the technical program, with scale, factorial praluction, fermenter volume often increases by 10 times, life Production cost will decline 37%-60%.
It should be noted that, the fermentation tank in the technical program is further equipped with air sterilization and filtering system (air cleaning system System), for aerobic liquid lucidum strain culture pure-blood ferment, air is that requirement is cleaned into sterile condition, nature air Contain countless dusts and microorganism, it is necessary to pass through a series of purified treatments, could enter fermentation tank as filtrated air.Nothing Effect of the bacterium air in fermenting and producing：One is to provide oxygen, and two be to promote to cultivate liquid to stir, and mycelium is in the medium not Disconnected mixing, improves the absorption and operation to nutriment, three be the uniform diffusion of maintenance metabolism product, and four are to maintain fermentation process Positive pressure operation, prevent microbiological contamination.
The air handled by air sterilization and filtering system reaches：Sterile, free from dust, free from admixture, anhydrous, oil-free, pressure Etc. several indexs.
Through experimental study, using solid medium and Liquid Strain of Ganoderma Lucidum culture medium prescription (embodiment 1-2) of the present invention Using effect is shown in Table 1.
In summary, The present invention reduces incubation step, shorten mycelium growing period, shorten growth cycle, contamination rate it is low, Cost is low, and yield is high.
It should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention and it is unrestricted, although with reference to preferable The present invention is described in detail embodiment, it will be understood by those within the art that, can be to technology of the invention Scheme is modified or equivalent substitution, and without departing from the spirit and scope of technical solution of the present invention, it all should cover in this hair Among bright right.

Claims (7)

1. a kind of Liquid Strain of Ganoderma Lucidum culture medium prescription, it is characterised in that including：Peptone 6g, glucose 15g, magnesium sulfate 0.5g, potassium dihydrogen phosphate 0.5g, farina 1g, corn, tapioca 3g, vitamin B110mg, agar 1g, pure water 1000mL。
2. a kind of Liquid Strain of Ganoderma Lucidum culture medium prescription, it is characterised in that including：Peptone 250g, glucose 1000g, sulfuric acid Magnesium 40g, potassium dihydrogen phosphate 20g, farina 100g, corn, tapioca 200g, vitamin B1600mg, agar 100g, Defoamer 5g, pure water 60L.
3. a kind of Liquid Strain of Ganoderma Lucidum cultural method, it is characterised in that comprise the following steps：
(1) prepare the triangular flask of multiple 500ml capacity, 200ml/ bottles are respectively charged into multiple triangular flasks by claim 1 or 2 Multiple triangular flasks are placed on high steam pot sterilizing, then by culture medium made from described Liquid Strain of Ganoderma Lucidum culture medium prescription Take out cooling；
(2) ganoderma lucidum slant tube parent species one are taken, 1 piece of picking broad bean size or 2 ferfas kinds are accessed in each triangular flask, Ran Houjing To 24 hours；
After (3) 24 hours, multiple triangular flasks are put into rotary shaker, cultivated on shaking table, shaking table culture 6 days, bottle is made Liquid spawn.
4. Liquid Strain of Ganoderma Lucidum cultural method as claimed in claim 3, it is characterised in that：High steam described in step (1) Pot sterilizing is specially high-temperature sterilization 30 minutes under the conditions of 120 DEG C -126 DEG C.
5. Liquid Strain of Ganoderma Lucidum cultural method as claimed in claim 3, it is characterised in that：The rotation of shaking table described in step (3) Rotary speed is in 120-130r/min, and temperature is 25 DEG C-30 DEG C.
6. Liquid Strain of Ganoderma Lucidum cultural method as claimed in claim 3, it is characterised in that：Also include following step after step (3) Suddenly：
(4) prepare the triangular flask of Large Copacity, vial of liquid strain made from step (3) is accessed into big triangular flask, liter/bottle of loading amount 4 Liquid spawn culture medium, rotary shaker is put into by the triangular flask of the Large Copacity, is cultivated 36 hours on shaking table；
(5) liter/bottle liquid spawn of loading amount 4 is accessed into 60 liters of stainless steel air blow tank, cultivated in the air blow tank 36 hours, Liquid Strain of Ganoderma Lucidum is made；
(6) by the Liquid Strain of Ganoderma Lucidum in fermentation tank, the sack portion for accessing ganoderma lucidum solid cultivating in bag strain is injected with syringe Position, carries out large-scale production.
7. Liquid Strain of Ganoderma Lucidum cultural method as claimed in claim 6, it is characterised in that：In step (5), aerobic fementation is supplied The air of tank is to pass through the filtrated air after a series of purified treatments of air sterilization and filtering system.
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