CN104017841A - Technique for producing truffle polysaccharide by truffle liquid fermentation - Google Patents
Technique for producing truffle polysaccharide by truffle liquid fermentation Download PDFInfo
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- CN104017841A CN104017841A CN201410263701.4A CN201410263701A CN104017841A CN 104017841 A CN104017841 A CN 104017841A CN 201410263701 A CN201410263701 A CN 201410263701A CN 104017841 A CN104017841 A CN 104017841A
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Abstract
The invention relates to an edible fungus truffle liquid fermentation technique, belonging to the technical field of bioengineering. The technique solves the problems of long period, high cost and the like in the artificial truffle culture process. The technique comprises the following steps: inoculating a truffle strain into a PDA (potato dextrose agar) culture medium to culture at 22-27 DEG C for 5-10 days; transferring into a triangular flask filled with a liquid culture medium, and culturing in an oscillating table at the rotation speed of 100-180 r/min at 22-27 DEG C for 5-8 days to obtain a liquid strain; inoculating the liquid strain into a fermentation tank filled with a fermentation culture medium, and carrying out fermentation culture at the mixing speed of 200 r/min at 22-27 DEG C under the conditions of dissolved oxygen 50% and pH value 6.0-8.0 for 5-8 days to obtain truffle mycelia, and extracting the truffle mycelia to obtain the truffle polysaccharide. The technique lowers the production cost, obviously enhances the truffle extracellular and intracellular polysaccharide yield by using the specific culture medium, and is beneficial to industrialized fermentation production.
Description
Technical field
The present invention relates to a kind of edible mushrooms ferfas liquid fermentation process, belong to technical field of bioengineering.
Background technology
Ferfas and caviare, pate de foie gras, be called as three large treasures America and Europe, the leader of European countries, is popular food in international connection, is universally acknowledged top food.The ascoma of ferfas is spherical in shape, semisphere or bulk, and diameter is generally at 1.5-12 centimetre, after ripe be brown, sorrel to Vandyke brown, base portion often caves in, surperficial cloth is with little wart not of uniform size.Ferfas is rich in 17 seed amino acids, 8 kinds of VITAMIN, appropriate protein and androsterone, sterol, sphingolipid, lipid acid, amino acid and trace elements etc. more than 50 are planted physiologically active ingredient, according to the study, truffle polysaccharides is as a new fungus polysaccharide, toxicity is low, good water solubility, function of tumor inhibition are obvious, is the medicine of antitumor immunotherapy.
Ferfas grows in underground medicine-food two-purpose fungi as a kind of, at occurring in nature, require the environment that grows in alkaline soil, has a moderate climate, simultaneously, with the Root Symbionts of the seeds such as Oak Tree, robur, harsh growth conditions has directly caused its natural output, and supply falls short of demand.In order to make up the deficiency of the natural output of ferfas, there are many scholars to carry out the research of semi-artificial simulation cultivation; but artificial culture exists the cycle long; generally need 7-9; need to expend a large amount of manpowers and physics; cost is high; and be subject to environmental influence large; limited to a certain extent its exploitation; and adopt liquid fermentation technology, be the effective ways of current deep development edible fungus active composition; and success has been accomplished scale production at rare edible medicinal fungus such as glossy ganoderma, mushrooms, but the research of ferfas liquid fermenting report is less at present.
Summary of the invention
The object of the invention is to for the deficiencies in the prior art, provide a kind of fermentation costs low, the technique that the ferfas liquid fermenting that mycelia production efficiency is high is produced truffle polysaccharides.
The above-mentioned object of the present invention is achieved through the following technical solutions: a kind of ferfas liquid fermenting is produced the technique of truffle polysaccharides, and described technique comprises the steps:
S1, slant strains activation culture: ferfas bacterial classification is inoculated on PDA substratum, carries out the cultivation of slant strains, culture temperature 22-27 ℃, incubation time 5-10 days;
The preparation of S2, liquid spawn: the slant strains having activated in picking step S1, transferred and be equipped with in the triangular flask of liquid nutrient medium, be placed in shaking table cultivation and make liquid spawn, culture condition is: temperature is 22-27 ℃, shaking speed is 100-180r/min, and incubation time is 5-8 days; The moiety of described liquid nutrient medium is: glucose: 15-25g/L, yeast: 1.5-2.5g/L, malt extract: 2.5-7.5g/L, KH
2pO
4: 0.5-1.5g/L, MgSO
4.7H
2o:0.25-0.75g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water, pH value is 6.0-8.0;
S3, liquid fermentation and culture: the liquid-spawn inoculation making in step S2 is carried out to fermentation culture in the fermentor tank of fermention medium and obtains ferfas mycelium to being equipped with, through extracting, make truffle polysaccharides; Wherein, fermentation condition is: stirring velocity is 200r/min, and dissolved oxygen is that 50%, pH value is 6.0-8.0, temperature 22-27 ℃, time 5-8 days; The moiety of described fermention medium is: glucose: 15-25g/L, malt extract: 2.5-7.5g/L, KH
2pO
4: 0.5-1.5g/L, MgSO
4.7H
2o:0.25-0.75g/L, lactoalbumin hydrolysate: 25-35g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water.
If shaking speed is too slow in the processing step S2 of ferfas liquid fermenting production truffle polysaccharides of the present invention, the dissolved oxygen in substratum is just not enough, will soon affect mycelial growth.If incubation time is too short, mycelia is no growth also, and the output of truffle polysaccharides is extremely low, and the oversize substratum nutrition of incubation time is inadequate, and the easy self-dissolving of mycelia affects the yield of final truffle polysaccharides on the contrary.And the too high or too low growth that all can suppress mycelia of temperature, pH of cultivating, thereby affect the yield of truffle polysaccharides.
Dissolved oxygen in step S3, pH value, leavening temperature, fermentation time all directly affect the growth of ferfas, too high or the too low yield that all can reduce final product, therefore, in the technique of ferfas liquid fermenting production truffle polysaccharides of the present invention, by strict control of the fermentation condition in step S3, be stirring velocity 200r/min, dissolved oxygen 50%, pH value 6.0-8.0, temperature 22-27 ℃, time 5-8 days.
As preferably, at above-mentioned ferfas liquid fermenting, produce in the technique of truffle polysaccharides, the ferfas bacterial classification described in step S1 is one or more in India truffle (TuberIndicum), the primary ferfas in Taiwan (TuberFormosanum), the white ferfas (TuberMagnatum) of Italy, French black truffle (TuberMelanosporum), false depression ferfas (Tuber Pseudoexcavatum).These ferfas bacterial classifications all can be bought and be obtained by commercial sources.
As preferably, at above-mentioned ferfas liquid fermenting, to produce in the technique of truffle polysaccharides, the moiety of the liquid nutrient medium described in step S2 is: glucose: 20g/L, yeast: 2.0g/L, malt extract: 5.0g/L, KH
2pO
4: 1.0g/L, MgSO
4.7H
2o:0.5g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water.
As preferably, at above-mentioned ferfas liquid fermenting, to produce in the technique of truffle polysaccharides, the moiety of the fermention medium described in step S3 is: glucose: 20g/L, malt extract: 5.0g/L, KH
2pO
4: 1.0g/L, MgSO
4.7H
2o:0.5g/L, VITMAIN B1: 100 μ g/L, lactoalbumin hydrolysate: 30g/L, surplus is distilled water.
As preferably, at above-mentioned ferfas liquid fermenting, to produce in the technique of truffle polysaccharides, the inoculum size described in step S3 is 200-800mg/L.
As a reference: described PDA nutrient agar is: MgSO
4.7H
2o:0.15g/L, KH
2pO
4: 0.30g/L, glucose: 20g/L, VITMAIN B1: 0.05g/L, potato: 200g/L.Described PDA nutrient agar can be bought and be obtained by commercial sources.
Compared with prior art, tool of the present invention has the following advantages:
(1), the present invention avoided extracting the unfavorable factor such as polysaccharide process Raw is limited from expensive kames, greatly reduces production cost.
(2), the present invention is by specific substratum, effectively Promote cell's growth, improves biomass, significantly improves the output of the outer and intracellular polyse of ferfas born of the same parents, can reach 2.5g/L, is conducive to industrial fermentation production.
Embodiment
Be below specific embodiments of the invention, technical scheme of the present invention is further described, but the present invention is not limited to these embodiment.
Embodiment 1
Slant strains activation culture: ferfas bacterial classification is inoculated on PDA substratum, carries out the cultivation of slant strains, 23 ℃ of culture temperature, incubation time 7 days;
The preparation of liquid spawn: the good slant strains of the above-mentioned activation of picking, transferred and be equipped with in the triangular flask of liquid nutrient medium, be placed in shaking table cultivation and make liquid spawn, culture condition is: temperature is 23 ℃, shaking speed is 120r/min, and incubation time is 6 days; The moiety of described liquid nutrient medium is: glucose: 20g/L, yeast: 2.0g/L, malt extract: 5.0g/L, KH
2pO
4: 1.0g/L, MgSO
4.7H
2o:0.50g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water, pH value is 6.0-8.0;
Liquid fermentation and culture: the above-mentioned liquid-spawn inoculation making is carried out to fermentation culture in the fermentor tank of fermention medium and obtains ferfas mycelium to being equipped with, initial inoculum is 500mg/L, fermentation condition is: stirring velocity is 200r/min, dissolved oxygen is 50%, initial pH value is 6.0,23 ℃ of temperature, 7 days time; The moiety of described fermention medium is: glucose: 20g/L, malt extract: 5.0g/L, KH
2pO
4: 1.0g/L, MgSO
4.7H
2o:0.50g/L, lactoalbumin hydrolysate: 30g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water;
The above-mentioned ferfas mycelium making is dried to constant weight at 50 ℃, grind into powder, get 100mg, with ethanol lixiviate 30min at 60 ℃ of 15mL85%, repeat twice, remove the interference components such as monosaccharide and disaccharide, oligose, lixiviate 1h in boiling water bath again, the centrifugal 15min of 5000r/min, gets supernatant liquor and is settled to 100mL, and it is 2.6g/L that the vitriol oil-phynol method is measured polysaccharide content.
Embodiment 2
Slant strains activation culture: ferfas bacterial classification is inoculated on PDA substratum, carries out the cultivation of slant strains, 25 ℃ of culture temperature, incubation time 6 days;
The preparation of liquid spawn: the good slant strains of the above-mentioned activation of picking, transferred and be equipped with in the triangular flask of liquid nutrient medium, be placed in shaking table cultivation and make liquid spawn, culture condition is: temperature is 24 ℃, shaking speed is 120r/min, and incubation time is 5 days; The moiety of described liquid nutrient medium is: glucose: 25g/L, yeast: 1.8g/L, malt extract: 3.0g/L, KH
2pO
4: 1.2g/L, MgSO
4.7H
2o:0.60g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water, pH value is 6.0-8.0;
Liquid fermentation and culture: the above-mentioned liquid-spawn inoculation making is carried out to fermentation culture in the fermentor tank of fermention medium and obtains ferfas mycelium to being equipped with, initial inoculum is 500mg/L, fermentation condition is: stirring velocity is 200r/min, dissolved oxygen is 50%, initial pH value is 6.0,24 ℃ of temperature, 5 days time; The moiety of described fermention medium is: glucose: 25g/L, malt extract: 3.0g/L, KH
2pO
4: 1.2g/L, MgSO
4.7H
2o:0.60g/L, lactoalbumin hydrolysate: 28g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water;
The above-mentioned ferfas mycelium making is dried to constant weight at 50 ℃, grind into powder, get 100mg, with ethanol lixiviate 30min at 60 ℃ of 15mL85%, repeat twice, remove the interference components such as monosaccharide and disaccharide, oligose, lixiviate 1h in boiling water bath again, the centrifugal 15min of 5000r/min, gets supernatant liquor and is settled to 100mL, and it is 2.4g/L that the vitriol oil-phynol method is measured polysaccharide content.
Embodiment 3
Slant strains activation culture: ferfas bacterial classification is inoculated on PDA substratum, carries out the cultivation of slant strains, 22 ℃ of culture temperature, incubation time 8 days;
The preparation of liquid spawn: the good slant strains of the above-mentioned activation of picking, transferred and be equipped with in the triangular flask of liquid nutrient medium, be placed in shaking table cultivation and make liquid spawn, culture condition is: temperature is 26 ℃, shaking speed is 170r/min, and incubation time is 7 days; The moiety of described liquid nutrient medium is: glucose: 18g/L, yeast: 2.3g/L, malt extract: 6.0g/L, KH
2pO
4: 0.7g/L, MgSO
4.7H
2o:0.30g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water, pH value is 6.0-8.0;
Liquid fermentation and culture: the above-mentioned liquid-spawn inoculation making is carried out to fermentation culture in the fermentor tank of fermention medium and obtains ferfas mycelium to being equipped with, initial inoculum is 500mg/L, fermentation condition is: stirring velocity is 200r/min, dissolved oxygen is 50%, initial pH value is 7.0,26 ℃ of temperature, 6 days time; The moiety of described fermention medium is: glucose: 15g/L, malt extract: 5.5g/L, KH
2pO
4: 0.8g/L, MgSO
4.7H
2o:0.30g/L, lactoalbumin hydrolysate: 32g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water;
The above-mentioned ferfas mycelium making is dried to constant weight at 50 ℃, grind into powder, get 100mg, with ethanol lixiviate 30min at 60 ℃ of 15mL85%, repeat twice, remove the interference components such as monosaccharide and disaccharide, oligose, lixiviate 1h in boiling water bath again, the centrifugal 15min of 5000r/min, gets supernatant liquor and is settled to 100mL, and it is 2.5g/L that the vitriol oil-phynol method is measured polysaccharide content.
The polysaccharide content that in prior art, the preparation method of liquid deep fermentation for producing truffle polysaccharide makes is generally 0.91-1.60g/L, the visible technique by ferfas liquid fermenting production truffle polysaccharides of the present invention has obviously improved the output of polysaccharide, and greatly reduce production cost, be conducive to industrial fermentation and produce.
Specific embodiment described herein is only to the explanation for example of the present invention's spirit.Those skilled in the art can make various modifications or supplement or adopt similar mode to substitute described specific embodiment, but can't depart from spirit of the present invention or surmount the defined scope of appended claims.
Although the present invention has been made a detailed description and has quoted as proof some specific embodiments, to those skilled in the art, only otherwise it is obvious leaving that the spirit and scope of the present invention can make various changes or revise.
Claims (5)
1. ferfas liquid fermenting is produced a technique for truffle polysaccharides, it is characterized in that, described technique comprises the steps:
S1, slant strains activation culture: ferfas bacterial classification is inoculated on PDA substratum, carries out the cultivation of slant strains, culture temperature 22-27 ℃, incubation time 5-10 days;
The preparation of S2, liquid spawn: the slant strains having activated in picking step S1, transferred and be equipped with in the triangular flask of liquid nutrient medium, be placed in shaking table cultivation and make liquid spawn, culture condition is: temperature is 22-27 ℃, shaking speed is 100-180r/min, and incubation time is 5-8 days; The moiety of described liquid nutrient medium is: glucose: 15-25g/L, yeast: 1.5-2.5g/L, malt extract: 2.5-7.5g/L, KH
2pO
4: 0.5-1.5g/L, MgSO
4.7H
2o:0.25-0.75g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water, pH value is 6.0-8.0;
S3, liquid fermentation and culture: the liquid-spawn inoculation making in step S2 is carried out to fermentation culture in the fermentor tank of fermention medium and obtains ferfas mycelium to being equipped with, through extracting, make truffle polysaccharides; Wherein, fermentation condition is: stirring velocity is 200r/min, and dissolved oxygen is that 50%, pH value is 6.0-8.0, temperature 22-27 ℃, time 5-8 days; The moiety of described fermention medium is: glucose: 15-25g/L, malt extract: 2.5-7.5g/L, KH
2pO
4: 0.5-1.5g/L, MgSO
4.7H
2o:0.25-0.75g/L, lactoalbumin hydrolysate: 25-35g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water.
2. ferfas liquid fermenting according to claim 1 is produced the technique of truffle polysaccharides, it is characterized in that, the ferfas bacterial classification described in step S1 is one or more in India truffle (Tuber Indicum), the primary ferfas in Taiwan (TuberFormosanum), the white ferfas (TuberMagnatum) of Italy, French black truffle (TuberMelanosporum), false depression ferfas (TuberPseudoexcavatum).
3. ferfas liquid fermenting according to claim 1 is produced the technique of truffle polysaccharides, it is characterized in that, the moiety of the liquid nutrient medium described in step S2 is: glucose: 20g/L, yeast: 2.0g/L, malt extract: 5.0g/L, KH
2pO
4: 1.0g/L, MgSO
4.7H
2o:0.5g/L, VITMAIN B1: 100 μ g/L, surplus is distilled water.
4. ferfas liquid fermenting according to claim 1 is produced the technique of truffle polysaccharides, it is characterized in that, the moiety of the fermention medium described in step S3 is: glucose: 20g/L, malt extract: 5.0g/L, KH
2pO
4: 1.0g/L, MgSO
4.7H
2o:0.5g/L, VITMAIN B1: 100 μ g/L, lactoalbumin hydrolysate: 30g/L, surplus is distilled water.
5. ferfas liquid fermenting according to claim 1 is produced the technique of truffle polysaccharides, it is characterized in that, the inoculum size described in step S3 is 200-800mg/L.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104824768A (en) * | 2015-04-14 | 2015-08-12 | 天津市林业果树研究所 | Edible fungus beverage preparation method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
|---|
| 朱伶俐: "中国块菌深层发酵生产块菌多糖的初步探索", 《中国优秀硕士学位论文全文数据库（农业科技辑）》 * |
| 马双双: "粒毛盘菌YM218胞外多糖深层发酵及抗肿瘤活性研究", 《中国优秀硕士学位论文全文数据库（基础科学辑）》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104824768A (en) * | 2015-04-14 | 2015-08-12 | 天津市林业果树研究所 | Edible fungus beverage preparation method |
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