CN104498367A - Medium for cordyceps militaris liquid deep fermentation and use method thereof - Google Patents
Medium for cordyceps militaris liquid deep fermentation and use method thereof Download PDFInfo
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Abstract
The invention discloses a medium for cordyceps militaris liquid deep fermentation. The medium comprises 10-30g of cane sugar, 1-5g of corn steep liquor, 4-100g of fermenting mash protein residue, 0-4g of peptone, 0-4g of beef extract, 0.5-2.0g of MgSO4.7H2O, 0.2-1.0g of KH2PO4, 1-5mg of vitamin B1 and 1000ml of water. The medium contains waste fermenting mash protein residue obtained by vitamin C fermentation, the fermenting mash protein residue contains mycoprotein of bacillus megatherium and ketogulonigenium vulgare, and the mycoprotein is similar to silkworm pupa protein and thus the mycoprotein can completely or partially replace beef extract and peptone in the medium. The medium can improve a cordyceps militaris mycelium fermentation yield, realize resource utilization of the waste fermenting mash protein residue obtained by a vitamin C fermentation industry and effectively reduce a production cost. The invention also provides a use method of the medium for cordyceps militaris liquid deep fermentation.
Description
Technical field
The present invention relates to edible mushrooms and cultivate field, particularly relate to a kind of substratum for Cordyceps militaris submerged fermentation and using method thereof.
Background technology
Cordyceps militaris (L.) Link. formal name used at school Cordyceps militaris (L.) Link. (
cordyceps militaris), it and Cordyceps of Qinghai Province belong to Mycophyta Ascomycotina Cordyceps, therefore, are called belonging to together " brother " of Cordyceps sinensis by expert.The perfect stage of Cordyceps militaris (L.) Link. life history comprises mycelial growth and sporophore growth.The insect larvaes such as lepidopteran, Coleoptera, Diptera or pupa infect the spore of Paecilomyces varioti, and when envrionment conditions is suitable for, spore just sprouts formation mycelium, and the nutrition in mycelium absorption worm (pupa) body is as its material grown and the energy.After mycelia decomposes the various tissue in worm (pupa) body and organ, hyphostoma development transfers reproductive growth to by nourishing and growing, mycelium twist together gradually be differentiated to form sclerotium and pass that the external top becoming safran or orange is slightly expanded, in bar-shaped stroma (sporophore), the many heads from polypide of stroma, chest, caudad etc. stretch out.
Wild Cordyceps militaris (L.) Link. is similar to traditional Cordyceps sinensis, very rare at occurring in nature.Most research is thought, Cordyceps militaris (L.) Link. is close with natural cordyceps on effective component, and its medicinal and edibleness can match in excellence or beauty with natural Cordyceps sinensis.The Cordyceps militaris (L.) Link. cellulose content of factorial praluction is higher than natural cordyceps, and the content of its protein, amino acid, VITAMIN and trace element has has all met or exceeded the level of natural cordyceps.Plant in Cordyceps reported more than 350, Cordyceps militaris (L.) Link. is the rare Cordyceps that can form a large amount of cordycepin.Because with Cordyceps sinensis sibship recently, effective constituent is quite even higher, and can artificial culture, Cordyceps militaris (L.) Link. becomes the Ideal Substitute of natural cordyceps.
The cultivation of Cordyceps militaris (L.) Link. mainly contains two kinds of modes, i.e. solid culture and liquid fermenting.People are devoted to the substitute of liquid submerged fermentation cultivation Cordyceps mycelium as Cordyceps sinensis always for a long time.The substratum of traditional Chinese caterpillar fungus liquid submerged fermentation mainly contains following several: 1. potato 200g(liquor), sucrose 20g, potassium primary phosphate 1g, magnesium sulfate 0.5g, peptone 5g, water 1000ml; 2. glucose 30g, lactoalbumin hydrolysate 5g, milk 100ml, beef soup 500ml, yeast extract paste 1g, potassium primary phosphate 1g, magnesium sulfate 0.2g, vitamin B complexes 1, water 1000ml; 3. glucose 20g, peptone 4g, extractum carnis 4g, potassium primary phosphate 1g, magnesium sulfate 0.5g, VITMAIN B1 2mg, water 1000ml.
Wild cordyceps militaris is at the silkworm chrysalis tumor growth containing a large amount of insect protein, but because reasons such as cost is high, operation difficulties, be difficult to adopt silkworm chrysalis or its component to do substratum under culture conditions, thus cause the problems such as cordyceps militaris plantation output and effective constituent all decline to a great extent.Therefore, a kind of novel culture medium for Cordyceps militaris submerged fermentation of exploitation is needed badly, with the demand of satisfied reality.
On the other hand, ascorbic industrial production adopts bio-fermentation process, gives off a large amount of waste fermented mash albumen slag.This waste eggshell white slag is beige, slightly thick, COD value 7-10 ten thousand mg/L, the macromolecular substance of fermented liquid in subsequent extracted step after ultra-filtration membrane retains more than remaining 100,000 dalton of vitamins C second step fermentation, main component is that the thalline of bacillus megaterium and raw ketone group 2-KLG bacterium, bacterial chip, residue substratum are as corn steep liquor, urea etc., wherein dry matter content is 10-15%, and crude protein content accounts for the 60-65% of dry-matter, therefore also known as mash albumen slag.Have that thalline breaks, thalline content overflows in the vitamin C bio fermentation later stage and be dissolved in the phenomenon in fermented liquid.
The invention provides a kind of substratum for Cordyceps militaris submerged fermentation and using method thereof, vitamin c fermenting waste eggshell white slag is added in Cordyceps militaris submerged fermentation culture medium, containing the tropina of bacillus megaterium with raw ketone group 2-KLG bacterium in this mash albumen slag, be similar to pupa albumen, therefore can extractum carnis and peptone in all or part of substitutive medium, both cordyceps mycelium fermentation production rate can have been made to obtain improve, the discarded mash albumen slag of vitamin c fermenting industry is given recycling again simultaneously, effectively reduce production cost.
Summary of the invention
The object of the present invention is to provide a kind of substratum for Cordyceps militaris submerged fermentation and using method thereof, this substratum effectively can improve the fermentation production rate of cordyceps mycelium, and effectively reduces production cost.
For realizing above-mentioned purpose of the present invention, the invention provides a kind of substratum for Cordyceps militaris submerged fermentation, being made up of following raw material: sucrose 10-30g, corn steep liquor 1-5g, mash albumen slag 4-100g, peptone 0-4g, extractum carnis 0-4g, MgSO
47H
2o 0.5-2.0g, KH
2pO
40.2-1.0g, VITMAIN B1 1-5mg, water 1000mL.
The present invention also provides a kind of using method of the substratum for Cordyceps militaris submerged fermentation, and concrete steps are as follows.
Step 1, substratum and sterilizing thereof and inoculation: carry out sterilizing to slack tank, breather line, air filter and discharge port by steam generator, pressure 0.1-0.15MPa, temperature 121-124 DEG C, continue sterilizing 40min.
Ferment tank is cultivated and shake-flask seed cultivates liquid nutrient medium used: sucrose 10-30g, corn steep liquor 1-5g, mash albumen slag 4-100g, peptone 0-4g, extractum carnis 0-4g, MgSO
47H
2o 0.5-2.0g, KH
2pO
40.2-1.0g, VITMAIN B1 1-5mg, water 1000mL.
Load after substratum preparation in fermentor tank, Intake Quantity is the 60-70% of fermentor tank capacity, then at pressure 0.12-0.15MPa, sterilizing 40min under temperature 121-126 DEG C of condition, start cold water circulating system to lower the temperature, when temperature drops to 25 DEG C inoculation liquid Cordyceps militaris spawn (this liquid spawn by Cordyceps militaris (L.) Link. (
cordyceps militaris) fermentation).
Step 2, culture condition control.
Fermentation tank pressure: when liquid fermentation and culture, if fermentor tank external and internal pressure is identical, extraneous miscellaneous bacteria can enter in fermentor tank and cause bacterium liquid inductance to contaminate; And the pressure of fermentor tank is too large, then affect dissolved oxygen and mycelial growth growth.In culturing process of the present invention, fermentor tank pressure-controlled is at 0.01-0.02MPa.
The control of foam: in liquid fermentation and culture mid-term, thalline enters vigorous period, and respiratory metabolism is vigorous, there will be foam, for preventing CO
2poisoning and miscellaneous bacteria infects, and will carry out froth breaking, its method can add a small amount of defoamer before sterilization in nutrient solution, and consumption is 0.05-0.1%.Also can adopt Trisun Oil R 80 or edible oil, to reduce surface tension, reduce formation of foam.
Air flow controls: when other factors is determined, air flow is directly proportional to dissolved oxygen coefficient, and different growth periods requires different to dissolved oxygen.In liquid fermentation and culture process of the present invention, the ventilation in fermentor tank controls at 1:0.2-1:0.4 m
3/ m
3/ min.
Stirring velocity: when liquid fermentation and culture, constantly stirs and various nutritive ingredient in liquid nutrient medium can be made to mix and unlikely precipitation, promotes gas and liquid comes into contact and exchange simultaneously; Stirring velocity is too high or too low is all unfavorable for that mycelial growth is grown.When the present invention cultivates, stirring velocity controls at 110-120rpm/min.
PH value controls: fermenting process adopts two-stage control pH strategy, and when namely starting, control ph is constant in 6.2, and after inoculation Cordyceps militaris bacterial classification, pH value constant control is 5.6 by 50h again.
Leavening temperature and cycle: leavening temperature controls at 18-24 DEG C, every grade of fermentation culture cycle within the scope of 96-120h, when every grade of fermentation culture cycle higher than 120h time, slowly or stop growing, nutrient solution becomes clarification to mycelial growth.
Step 3, mycelia detect: for guaranteeing fermented quality, grasp hypha fermentation and cultivate situation, must carry out interassay.Open discharge port, by rotary sample gently after in bacterium liquid access sterile chamber, leave standstill 5 minutes, if mycelium suspended power is good, then show that vitality is strong; If very easily precipitate, illustrate that mycelia is aging, vigor reduces.Then by the film-making of bacterium liquid, check mycelia purity under the microscope, if when detecting afterwards without miscellaneous bacteria, can continue to cultivate; When conidium almost comes off parent sporophore entirely, when sporophore quantity increases not obvious, illustrate that mycelium culture terminates, can mycelia collection be carried out.
Step 4, mycelia are collected: after liquid fermentation and culture terminates, open discharge port, take out nutrient solution, collect mycelia, then the mycelia of collection is placed on air seasoning place and dries in the shade by centrifuging.
Compared with prior art beneficial effect of the present invention.
The invention provides a kind of substratum for Cordyceps militaris submerged fermentation and using method thereof, add the discarded mash albumen slag of vitamin c fermenting in the medium, containing the tropina of bacillus megaterium with raw ketone group 2-KLG bacterium in this mash albumen slag, be similar to pupa albumen, therefore can extractum carnis and peptone in all or part of substitutive medium, both cordyceps mycelium fermentation production rate can have been made to obtain improve, the discarded mash albumen slag of vitamin c fermenting industry is given recycling again simultaneously, effectively reduce production cost.
Embodiment
The present invention is further described below in conjunction with specific embodiment.
Embodiment 1.
The invention provides a kind of substratum for Cordyceps militaris submerged fermentation, be made up of following raw material: sucrose 20g, corn steep liquor 2g, mash albumen slag 8g, peptone 2g, extractum carnis 1.5g, MgSO
47H
2o 0.8g, KH
2pO
40.5g, VITMAIN B1 2mg, water 1000mL.
The invention provides a kind of using method of the substratum for Cordyceps militaris submerged fermentation, concrete technology parameter is as follows.
In fermentor tank, substratum loading amount is 65%, then sterilizing 40min under temperature 124 DEG C of conditions, when substratum temperature drops to 25 DEG C, inoculate liquid Cordyceps militaris spawn.
In liquid fermentation and culture process, in fermentor tank, temperature controls at 22 DEG C, and the ventilation in fermentor tank controls at 1:0.3 m
3/ m
3/ min, stirring velocity controls at 110rpm/min, and earlier fermentation pH value controls 6.2, and after inoculation Cordyceps militaris bacterial classification, pH value constant control is 5.6 by 50h again, carries out mycelia collection during fermentation 96h.
The mycelia yield that substratum of the present invention ferments is adopted to reach 0.40%, 29% is improve than mycelia yield that conventional medium obtains (0.31%), the mycelial cordycepin content of substratum of the present invention is 0.165%, obtains mycelial cordycepin content (0.125%) improve 24.2% than conventional medium.Note: mycelia yield (%)=mycelium dry weight (kg) ÷ puts tank secondary fermentation liquid long-pending × 100%.
Embodiment 2.
The invention provides a kind of substratum for Cordyceps militaris submerged fermentation, be made up of following raw material: sucrose 20g, corn steep liquor 2g, mash albumen slag 20g, extractum carnis 0.5g, MgSO
47H
2o 0.8g, KH
2pO
40.5g, VITMAIN B1 2mg, water 1000mL.
The invention provides a kind of using method of the substratum for Cordyceps militaris submerged fermentation, concrete technology parameter is as follows.
In fermentor tank, substratum loading amount is 60%, then sterilizing 40min under temperature 122 DEG C of conditions, when substratum temperature drops to 25 DEG C, inoculate liquid Cordyceps militaris spawn.
In liquid fermentation and culture process, in fermentor tank, temperature controls at 22 DEG C, and the ventilation in fermentor tank controls at 1:0.3 m
3/ m
3/ min, stirring velocity controls at 110rpm/min, and earlier fermentation pH value controls 6.2, and after inoculation Cordyceps militaris bacterial classification, pH value constant control is 5.6 by 50h again, carries out mycelia collection during fermentation 96h.
The mycelia yield that substratum of the present invention ferments is adopted to reach 0.37%, 19.3% is improve than mycelia yield that conventional medium obtains (0.31%), the mycelial cordycepin content of substratum of the present invention is 0.147%, obtains mycelial cordycepin content (0.125%) improve 17.6% than conventional medium.
Claims (4)
1. for a substratum for Cordyceps militaris submerged fermentation, be made up of following raw material: sucrose 10-30g, corn steep liquor 1-5g, mash albumen slag 4-100g, peptone 0-4g, extractum carnis 0-4g, MgSO
47H
2o 0.5-2.0g, KH
2pO
40.2-1.0g, VITMAIN B1 1-5mg, water 1000.
2., as claimed in claim 1 for the substratum of Cordyceps militaris submerged fermentation, it is characterized in that, be made up of following raw material: sucrose 20g, corn steep liquor 2g, mash albumen slag 8g, peptone 2g, extractum carnis 1.5g, MgSO
47H
2o 0.8g, KH
2pO
40.5g, VITMAIN B1 2mg, water 800mL.
3., as claimed in claim 2 for the substratum of Cordyceps militaris submerged fermentation, it is characterized in that, be made up of following raw material: sucrose 20g, corn steep liquor 2g, mash albumen slag 20g, extractum carnis 0.5g, MgSO
47H
2o 0.8g, KH
2pO
40.5g, VITMAIN B1 2mg, water 1000mL.
4., as claimed in claim 1 for the using method of the substratum of Cordyceps militaris submerged fermentation, it is characterized in that, concrete steps are:
Step 1, substratum and sterilizing thereof and inoculation: sterilizing is carried out to slack tank, breather line, air filter and discharge port by steam generator, pressure 0.1-0.15MPa, temperature 121-124 DEG C, continue sterilizing 40min;
Ferment tank cultivation and shake-flask seed are cultivated liquid nutrient medium used and are: sucrose 10-30g, corn steep liquor 1-5g, mash albumen slag 4-100g, peptone 0-4g, extractum carnis 0-4g, MgSO
47H
2o 0.5-2.0g, KH
2pO
40.2-1.0g, VITMAIN B1 1-5mg, water 1000mL;
Load in fermentor tank after described substratum preparation, Intake Quantity is the 60-70% of fermentor tank capacity, then sterilizing 40min under pressure 0.12-0.15MPa, temperature 121-126 DEG C condition, start cold water circulating system to lower the temperature, when temperature drops to 25 DEG C, inoculate liquid Cordyceps militaris spawn;
Step 2, culture condition control:
Fermentation tank pressure: in culturing process, fermentor tank pressure-controlled is at 0.01-0.02MPa;
The control of foam: add the one in defoamer, Trisun Oil R 80, edible oil before sterilization in nutrient solution, to reduce formation of foam, described defoamer consumption is 0.05-0.1%;
Air flow controls: the ventilation in fermentor tank controls at 1:0.2-1:0.4 m
3/ m
3/ min;
Stirring velocity: during cultivation, stirring velocity controls at 110-120rpm/min;
PH value controls: fermenting process adopts two-stage control pH strategy, and when namely starting, control ph is constant in 6.2, and after inoculation Cordyceps militaris bacterial classification, pH value constant control is 5.6 by 50h again;
Leavening temperature and cycle: leavening temperature controls at 18-24 DEG C, every grade of fermentation culture cycle is at 96-120h;
Step 3, mycelia detect;
Step 4, mycelia are collected.
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Cited By (1)
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| CN104756767A (en) * | 2015-04-29 | 2015-07-08 | 天津农学院 | High-density pleurotus eryngii liquid strain and fermentation method thereof |
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2014
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Patent Citations (6)
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| AU2013248822A1 (en) * | 2012-04-16 | 2014-06-12 | Institute Of Agricultural Resources And Environment, Shandong Academy Of Agricultural Sciences | Method for producing plasmin by liquid fermentation of Cordyceps militaris |
| CN103858673A (en) * | 2014-03-18 | 2014-06-18 | 鲁东大学 | Method for planting cordyceps militaris by beer residue |
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