CN104017841B - The technique that a kind of truffles liquid fermentation produces truffle polysaccharides - Google Patents
The technique that a kind of truffles liquid fermentation produces truffle polysaccharides Download PDFInfo
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- CN104017841B CN104017841B CN201410263701.4A CN201410263701A CN104017841B CN 104017841 B CN104017841 B CN 104017841B CN 201410263701 A CN201410263701 A CN 201410263701A CN 104017841 B CN104017841 B CN 104017841B
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Abstract
The present invention relates to a kind of edible fungi truffles liquid fermentation process, belong to technical field of bioengineering.Described technique solves artificial culture truffle cycle length, high in cost of production problem, its concrete steps: truffles strain is inoculated in PDA culture medium and cultivates 5 10 days at 22 27 DEG C；Then transferring into the triangular flask equipped with fluid medium, be placed in the shaking table of rotating speed 100 180r/min cultivation and obtain liquid spawn in 58 days, temperature is 22 27 DEG C；By liquid-spawn inoculation to obtaining truffles mycelium equipped with the fermentation tank of fermentation medium carries out fermentation culture, fermentation condition: mixing speed 200r/min, dissolved oxygen 50%, pH value 6.0 8.0, temperature 22 27 DEG C, 58 days time, the extracted prepared truffle polysaccharides of truffles mycelium.Described technique reduces production cost, by specific culture medium, significantly improves outside truffles born of the same parents and the yield of intracellular polysaccharide, beneficially industrial fermentation produce.
Description
Technical field
The present invention relates to a kind of edible fungi truffles liquid fermentation process, belong to biological engineering skill
Art field.
Background technology
Truffles and caviar, goose liver catsup, be referred to as three big treasures America and Europe, in Europe state
Family leader is popular food in international association, is universally acknowledged top food.
The ascocarp of truffles is spherical in shape, hemispherical or bulk, diameter typically at 1.5-12 centimetre,
Being brown, bronzing to dark brown after maturation, base portion often caves in, and surface cloth is with size not
The little wart of one.Truffles rich in 17 kinds of aminoacid, 8 kinds of vitamin, appropriate protein,
And androsterone, sterol, sphingolipid, fatty acid, aminoacid and trace element etc. more than 50 plant
Physiologically active ingredient, it was found that truffle polysaccharides is as a new fungus polysaccharide, toxicity
Low, good water solubility, function of tumor inhibition are obvious, are the medicines of antitumor immunotherapy.
Truffles is as a kind of medicine-food two-purpose fungus being grown on underground, requirement in nature
The environment be grown on basic soil, having a moderate climate, simultaneously with the seeds such as Oak Tree, robur
Root Symbiont, harsh growth conditions directly results in its natural yield, and supply falls short of demand.
Many scholars are had to carry out semi-artificial simulation cultivation in order to make up the deficiency of the natural yield of truffles
Research, but to there is the cycle long for artificial culture, it is generally required to 7-9, needs to expend
Substantial amounts of manpower and physics, cost is high, and big by environmental influence, to a certain degree
On limit to its develop, and use liquid fermentation technology be current deep development food
With the effective ways of bacterium active component, and success is at the rare edible medicinal such as Ganoderma, Lentinus Edodes
Bacterium is accomplished scale production, but the research report of truffles liquid fermentation is less at present.
Summary of the invention
Present invention aims to the deficiencies in the prior art, it is provided that Yi Zhongfa
Ferment low cost, the technique that the truffles liquid fermentation that mycelia production efficiency is high produces truffle polysaccharides.
The above-mentioned purpose of the present invention is achieved through the following technical solutions: a kind of truffles liquid is sent out
Ferment produces the technique of truffle polysaccharides, and described technique comprises the steps:
S1, slant strains activation culture: truffles strain is inoculated in PDA culture medium,
Carry out the cultivation of slant strains, cultivation temperature 22-27 DEG C, incubation time 5-10 days；
S2, the preparation of liquid spawn: the slant strains activated in picking step S1,
Transferred equipped with in the triangular flask of fluid medium, be placed in shaking table cultivation and prepare liquid
Body strain, condition of culture is: temperature is 22-27 DEG C, and shaking speed is 100-180r/min,
Incubation time is 5-8 days；The constituent of described fluid medium is: glucose: 15-25
G/L, yeast: 1.5-2.5g/L, Fructus Hordei Germinatus extract: 2.5-7.5g/L, KH2PO4: 0.5-1.5
G/L, MgSO4.7H2O:0.25-0.75g/L, vitamin B1: 100 μ g/L, surplus
For distilled water, pH value is 6.0-8.0；
S3, liquid fermentation and culture: by the liquid-spawn inoculation prepared in step S2 to filling
Have in the fermentation tank of fermentation medium and carry out fermentation culture and obtain truffles mycelium, extracted
Prepare truffle polysaccharides；Wherein, fermentation condition is: mixing speed is 200r/min, dissolved oxygen
Being 50%, pH value is 6.0-8.0, temperature 22-27 DEG C, 5-8 days time；Described fermentation
The constituent of culture medium is: glucose: 15-25g/L, Fructus Hordei Germinatus extract: 2.5-7.5
G/L, KH2PO4: 0.5-1.5g/L, MgSO4.7H2O:0.25-0.75g/L, hydrolysis breast
Albumen: 25-35g/L, vitamin B1: 100 μ g/L, surplus is distilled water.
If shaking in truffles liquid fermentation of the present invention produces the processing step S2 of truffle polysaccharides
Bed rotating speed is too slow, and the dissolved oxygen in culture medium is the most not enough, will soon affect mycelial growth.
If incubation time is the shortest, mycelia also no growth, the yield of truffle polysaccharides is extremely low, and cultivates
Time oversize culture medium nutrition is inadequate, and the easy self-dissolving of mycelia affects final truffles many on the contrary
The yield of sugar.And the highest or the lowest growth that all can suppress mycelia of the temperature cultivated, pH,
Thus affect the yield of truffle polysaccharides.
Dissolved oxygen in step S3, pH value, fermentation temperature, fermentation time all directly affect
The growth of truffles, the too high or too low yield that all can reduce end product, therefore,
By the fermentation in step S3 in the technique of truffles liquid fermentation of the present invention production truffle polysaccharides
Condition strictly controls as mixing speed 200r/min, dissolved oxygen 50%, pH value 6.0-8.0,
Temperature 22-27 DEG C, 5-8 days time.
As preferably, in above-mentioned truffles liquid fermentation produces the technique of truffle polysaccharides, step
Truffles strain described in rapid S1 is that tuber indicum (TuberIndicum), Taiwan are primary
Truffles (TuberFormosanum), Italy truffles (TuberMagnatum) in vain,
France black truffle (TuberMelanosporum), false depression truffles (Tuber
Pseudoexcavatum) one or more in.These truffles strains all can pass through business
Approach is commercially available.
As preferably, in above-mentioned truffles liquid fermentation produces the technique of truffle polysaccharides, step
The constituent of the fluid medium described in rapid S2 is: glucose: 20g/L, yeast:
2.0g/L, Fructus Hordei Germinatus extract: 5.0g/L, KH2PO4: 1.0g/L, MgSO4.7H2O:
0.5g/L, vitamin B1: 100 μ g/L, surplus is distilled water.
As preferably, in above-mentioned truffles liquid fermentation produces the technique of truffle polysaccharides, step
The constituent of the fermentation medium described in rapid S3 is: glucose: 20g/L, Fructus Hordei Germinatus
Extract: 5.0g/L, KH2PO4: 1.0g/L, MgSO4.7H2O:0.5g/L, dimension
Raw element B1:100 μ g/L, lactoalbumin hydrolysate: 30g/L, surplus is distilled water.
As preferably, in above-mentioned truffles liquid fermentation produces the technique of truffle polysaccharides, step
Inoculum concentration described in rapid S3 is 200-800mg/L.
As reference: described PDA agar culture medium is: MgSO4.7H2O:0.15
G/L, KH2PO4: 0.30g/L, glucose: 20g/L, vitamin B1: 0.05g/L,
Rhizoma Solani tuber osi: 200g/L.Described PDA agar culture medium can be commercially available by commercial sources.
Compared with prior art, present invention have the advantage that
(1), present invention, avoiding extraction polysaccharide mistake from expensive kames
The unfavorable factor such as in journey raw material is limited, greatly reduces production cost.
(2), the present invention by specific culture medium, effectively facilitate cell growth, improve
Biomass, significantly improves outside truffles born of the same parents and the yield of intracellular polysaccharide, up to 2.5g/L, has
It is beneficial to industrial fermentation produce.
Detailed description of the invention
The following is the specific embodiment of the present invention, technical scheme is made further
Description, but the present invention is not limited to these embodiments.
Embodiment 1
Slant strains activation culture: truffles strain is inoculated in PDA culture medium, carries out
The cultivation of slant strains, cultivation temperature 23 DEG C, incubation time 7 days；
The preparation of liquid spawn: the slant strains that the above-mentioned activation of picking is good, transferred into
Equipped with in the triangular flask of fluid medium, it is placed in shaking table cultivation and prepares liquid spawn, training
Foster condition is: temperature is 23 DEG C, and shaking speed is 120r/min, and incubation time is 6
My god；The constituent of described fluid medium is: glucose: 20g/L, yeast: 2.0g/L,
Fructus Hordei Germinatus extract: 5.0g/L, KH2PO4: 1.0g/L, MgSO4.7H2O:0.50g/L,
Vitamin B1: 100 μ g/L, surplus is distilled water, and pH value is 6.0-8.0；
Liquid fermentation and culture: by above-mentioned prepared liquid-spawn inoculation to equipped with fermentation culture
Carrying out fermentation culture in the fermentation tank of base and obtain truffles mycelium, initial inoculum is 500
Mg/L, fermentation condition is: mixing speed is 200r/min, and dissolved oxygen is 50%, initially
PH value is 6.0, temperature 23 DEG C, 7 days time；The constituent of described fermentation medium
For: glucose: 20g/L, Fructus Hordei Germinatus extract: 5.0g/L, KH2PO4: 1.0g/L,
MgSO4.7H2O:0.50g/L, lactoalbumin hydrolysate: 30g/L, vitamin B1: 100 μ g/L,
Surplus is distilled water；
Above-mentioned prepared truffles mycelium is dried to constant weight at 50 DEG C, pulverizes
End, takes 100mg, extracts 30min with the ethanol of 15mL85% at 60 DEG C, weight
Multiple twice, remove the interference component such as monosaccharide and disaccharide, oligosaccharide, soak in boiling water bath
Carry 1h, 5000r/min and be centrifuged 15min, take supernatant and be settled to 100mL, dense sulfur
It is 2.6g/L that acid-phynol method measures polyoses content.
Embodiment 2
Slant strains activation culture: truffles strain is inoculated in PDA culture medium, carries out
The cultivation of slant strains, cultivation temperature 25 DEG C, incubation time 6 days；
The preparation of liquid spawn: the slant strains that the above-mentioned activation of picking is good, transferred into
Equipped with in the triangular flask of fluid medium, it is placed in shaking table cultivation and prepares liquid spawn, training
Foster condition is: temperature is 24 DEG C, and shaking speed is 120r/min, and incubation time is 5
My god；The constituent of described fluid medium is: glucose: 25g/L, yeast: 1.8g/L,
Fructus Hordei Germinatus extract: 3.0g/L, KH2PO4: 1.2g/L, MgSO4.7H2O:0.60g/L,
Vitamin B1: 100 μ g/L, surplus is distilled water, and pH value is 6.0-8.0；
Liquid fermentation and culture: by above-mentioned prepared liquid-spawn inoculation to equipped with fermentation culture
Carrying out fermentation culture in the fermentation tank of base and obtain truffles mycelium, initial inoculum is 500
Mg/L, fermentation condition is: mixing speed is 200r/min, and dissolved oxygen is 50%, initially
PH value is 6.0, temperature 24 DEG C, 5 days time；The constituent of described fermentation medium
For: glucose: 25g/L, Fructus Hordei Germinatus extract: 3.0g/L, KH2PO4: 1.2g/L,
MgSO4.7H2O:0.60g/L, lactoalbumin hydrolysate: 28g/L, vitamin B1: 100 μ g/L,
Surplus is distilled water；
Above-mentioned prepared truffles mycelium is dried to constant weight at 50 DEG C, pulverizes
End, takes 100mg, extracts 30min with the ethanol of 15mL85% at 60 DEG C, repeats
Twice, remove the interference component such as monosaccharide and disaccharide, oligosaccharide, extract in boiling water bath
1h, 5000r/min are centrifuged 15min, take supernatant and are settled to 100mL, and concentrated sulphuric acid-
It is 2.4g/L that phynol method measures polyoses content.
Embodiment 3
Slant strains activation culture: truffles strain is inoculated in PDA culture medium, carries out
The cultivation of slant strains, cultivation temperature 22 DEG C, incubation time 8 days；
The preparation of liquid spawn: the slant strains that the above-mentioned activation of picking is good, transferred into
Equipped with in the triangular flask of fluid medium, it is placed in shaking table cultivation and prepares liquid spawn, training
Foster condition is: temperature is 26 DEG C, and shaking speed is 170r/min, and incubation time is 7
My god；The constituent of described fluid medium is: glucose: 18g/L, yeast: 2.3g/L,
Fructus Hordei Germinatus extract: 6.0g/L, KH2PO4: 0.7g/L, MgSO4.7H2O:0.30g/L,
Vitamin B1: 100 μ g/L, surplus is distilled water, and pH value is 6.0-8.0；
Liquid fermentation and culture: by above-mentioned prepared liquid-spawn inoculation to equipped with fermentation culture
Carrying out fermentation culture in the fermentation tank of base and obtain truffles mycelium, initial inoculum is 500
Mg/L, fermentation condition is: mixing speed is 200r/min, and dissolved oxygen is 50%, initially
PH value is 7.0, temperature 26 DEG C, 6 days time；The constituent of described fermentation medium
For: glucose: 15g/L, Fructus Hordei Germinatus extract: 5.5g/L, KH2PO4: 0.8g/L,
MgSO4.7H2O:0.30g/L, lactoalbumin hydrolysate: 32g/L, vitamin B1: 100 μ g/L,
Surplus is distilled water；
Above-mentioned prepared truffles mycelium is dried to constant weight at 50 DEG C, pulverizes
End, takes 100mg, extracts 30min with the ethanol of 15mL85% at 60 DEG C, weight
Multiple twice, remove the interference component such as monosaccharide and disaccharide, oligosaccharide, soak in boiling water bath
Carry 1h, 5000r/min and be centrifuged 15min, take supernatant and be settled to 100mL, dense sulfur
It is 2.5g/L that acid-phynol method measures polyoses content.
In prior art, the preparation method of liquid deep fermentation for producing truffle polysaccharide prepares many
Sugar content is generally 0.91-1.60g/L, it is seen that produced by truffles liquid fermentation of the present invention
The technique of truffle polysaccharides significantly improves the yield of polysaccharide, and greatly reduces production cost,
Industrial fermentation is conducive to produce.
Specific embodiment described herein is only to present invention spirit theory for example
Bright.Those skilled in the art can be to described specific embodiment
Make various amendment or supplement or use similar mode to substitute, but without departing from the present invention
Spirit or surmount scope defined in appended claims.
It is embodied as although the present invention having been made a detailed description and having quoted some as proof
Example, but to those skilled in the art, as long as without departing from the spirit of the present invention
It is obvious for can making various changes with scope or revise.
Claims (4)
1. the technique that a truffles liquid fermentation produces truffle polysaccharides, it is characterised in that institute
State technique to comprise the steps:
S1, slant strains activation culture: truffles strain is inoculated in PDA culture medium,
Carry out the cultivation of slant strains, cultivation temperature 22-27 DEG C, incubation time 5-10 days；Institute
The truffles strain stated is tuber indicum, the primary truffles in Taiwan, Italy white truffles, France
One or more in black truffle, false depression truffles；
S2, the preparation of liquid spawn: the slant strains activated in picking step S1,
Transferred equipped with in the triangular flask of fluid medium, be placed in shaking table cultivation and prepare liquid
Body strain, condition of culture is: temperature is 22-27 DEG C, and shaking speed is 100-180r/min,
Incubation time is 5-8 days；The constituent of described fluid medium is: glucose:
15-25g/L, yeast: 1.5-2.5g/L, Fructus Hordei Germinatus extract: 2.5-7.5g/L, KH2PO4:
0.5-1.5g/L, MgSO4.7H2O:0.25-0.75g/L, vitamin B1: 100 μ g/L,
Surplus is distilled water, and pH value is 6.0-8.0；
S3, liquid fermentation and culture: by the liquid-spawn inoculation prepared in step S2 to filling
Have in the fermentation tank of fermentation medium and carry out fermentation culture and obtain truffles mycelium, extracted
Prepare truffle polysaccharides；Wherein, fermentation condition is: mixing speed is 200r/min, dissolved oxygen
Being 50%, pH value is 6.0-8.0, temperature 22-27 DEG C, 5-8 days time；Described fermentation
The constituent of culture medium is: glucose: 15-25g/L, Fructus Hordei Germinatus extract: 2.5-7.5g/L,
KH2PO4: 0.5-1.5g/L, MgSO4.7H2O:0.25-0.75g/L, lactoalbumin hydrolysate:
25-35g/L, vitamin B1: 100 μ g/L, surplus is distilled water.
Truffles liquid fermentation the most according to claim 1 produces the work of truffle polysaccharides
Skill, it is characterised in that the constituent of the fluid medium described in step S2 is:
Glucose: 20g/L, yeast: 2.0g/L, Fructus Hordei Germinatus extract: 5.0g/L, KH2PO4:
1.0g/L, MgSO4.7H2O:0.5g/L, vitamin B1: 100 μ g/L, surplus is for steaming
Distilled water.
Truffles liquid fermentation the most according to claim 1 produces the work of truffle polysaccharides
Skill, it is characterised in that the constituent of the fermentation medium described in step S3 is:
Glucose: 20g/L, Fructus Hordei Germinatus extract: 5.0g/L, KH2PO4: 1.0g/L,
MgSO4.7H2O:0.5g/L, vitamin B1: 100 μ g/L, lactoalbumin hydrolysate: 30g/L,
Surplus is distilled water.
Truffles liquid fermentation the most according to claim 1 produces the work of truffle polysaccharides
Skill, it is characterised in that the inoculum concentration described in step S3 is 200-800mg/L.
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| CN1995322A (en) * | 2006-12-27 | 2007-07-11 | 湖北工业大学 | Culture medium for liquid deep fermentation for producing truffle polysaccharide |
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| CN1995322A (en) * | 2006-12-27 | 2007-07-11 | 湖北工业大学 | Culture medium for liquid deep fermentation for producing truffle polysaccharide |
Non-Patent Citations (2)
| Title |
|---|
| 中国块菌深层发酵生产块菌多糖的初步探索;朱伶俐;《中国优秀硕士学位论文全文数据库（农业科技辑）》;20090115(第1期);第6-7页第2.1.3节，第6页第2.1.2节，第1页最后一段，第48页第4.1节 * |
| 粒毛盘菌YM218胞外多糖深层发酵及抗肿瘤活性研究;马双双;《中国优秀硕士学位论文全文数据库（基础科学辑）》;20130715(第7期);第20页最后一段 * |
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