CN101648029A - Method for curing and sterilizing culture media for large-scale cultivation of cordyceps militaris - Google Patents
Method for curing and sterilizing culture media for large-scale cultivation of cordyceps militaris Download PDFInfo
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- CN101648029A CN101648029A CN200810041692A CN200810041692A CN101648029A CN 101648029 A CN101648029 A CN 101648029A CN 200810041692 A CN200810041692 A CN 200810041692A CN 200810041692 A CN200810041692 A CN 200810041692A CN 101648029 A CN101648029 A CN 101648029A
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Abstract
The invention relates to a method for curing and sterilizing culture media, in particular to a method for curing and sterilizing culture media for large-scale cultivation of cordyceps militaris. The method for curing and sterilizing culture media for large-scale cultivation of cordyceps militaris is characterized in that the steps of curing and sterilizing are performed in combination; the temperature of curing and sterilization is between 100 and 135 DEG C; the time for curing and sterilization is between 40 and 50 minutes; curing and sterilization are synchronously performed in a double-doortunnel-type pressure steam sterilizer with a front door and a back door; the back door of the double-door tunnel-type pressure steam sterilizer is automatically locked when the front door is fed; andthe front door of the double-door tunnel-type pressure steam sterilizer is automatically locked when materials are discharged from the back door. The method solves the problem that the prior curing-sterilizing technology is incapable of treating culture media in large capacity and easy to cause bacterial infection resulting in large-scale mould in the process of treating culture matrixes.
Description
Technical field
The present invention relates to a kind of ripening and sterilizing methods of culture medium, the ripening and the sterilizing methods of particularly a kind of Cordyceps militaris (L.) Link. (Cordyceps Militaris) culture medium.
Background technology
Batch production large-scale production Cordyceps militaris (L.) Link. needs a large amount of culture medium.The culture medium of tissue culture's Cordyceps militaris (L.) Link. mainly is made up of rice, analysis for soybean powder, each proteinoid and organic substance institute compatibility.The very easily rotten and mouldy fermentation of the culture medium of tissue culture's Cordyceps militaris (L.) Link., so it must under aseptic state, just can enter into the operation of inoculation through after maturation process and the high-temperature pressurizing sterilization.The ripening of culture medium and the key technology of having sterilized into large-scale industrialized cultivation Cordyceps militaris (L.) Link..
Cultivate Cordyceps militaris (L.) Link. at present at home and abroad, owing to be small-scale test manufacture generally, and do not have quality standard, more do not have the mature experience of large-scale industrialized production (promptly producing Cordyceps militaris (L.) Link. more than 20 tons per year), so more employing horizontal circular steam sterilization pot carries out ripening and sterilization in the production process.This mode of production can not satisfy the requirement of large-scale industrialized production Cordyceps militaris (L.) Link. fully, and annual production can only reach about 5 tons at the most.Some scientific research institutions and production unit, no mass production techniques and working condition have adopted steam cure and autoclaving production method step by step, do not reach the requirement of thorough ripening of culture medium and sterilization, material pass on and seeded process in easier microbiological contamination, tend to cause large batch of going mouldy.
Summary of the invention
The objective of the invention is to propose a kind of ripening and sterilizing methods of culture medium of large-scale cultivation of cordyceps militaris; can't high capacity handle the problem of culture medium to solve present ripening and sterilizing methods; and culture medium microbiological contamination easily in ripening and sterilization process, cause large batch of problem of going mouldy.
For solving the problems of the technologies described above, the present invention adopts following technical scheme:
The ripening of the culture medium of large-scale cultivation of cordyceps militaris and sterilizing methods is characterized in that: maturation stage and sterilization steps merge carries out, and the temperature of ripening and sterilization is 100 ℃～135 ℃, and the temperature retention time of ripening and sterilization is 40min～50min.
Further, described ripening and sterilization are carried out in the double door tunnel type pressure steam sterilizer that two of front and back are arranged synchronously; The back door Qianmen double door tunnel type pressure steam sterilizer of locking voluntarily when locking, back door discharging voluntarily when described double door tunnel type pressure steam sterilizer is the Qianmen charging.
Double door tunnel type pressure steam sterilizer in the method for the invention, back door locking voluntarily when maximum characteristics are the Qianmen charging, Qianmen locking voluntarily during the discharging of back door, after having avoided material ripening sterilization, open simultaneously for two, form circulation of air, cause cross-contamination.Steriliser is the horizontal pressure steam sterilizer of SVW 105 types that is provided by Shanghai Huaxian Medical Nuclear Instruments Co., Ltd., and portion uses mechanical interlocking principle of the prior art within it, makes it reach a unlatching, and another is the result of locking voluntarily.
Further, after ripening and the sterilization, with culture medium send to that back door with described double door tunnel type pressure steam sterilizer is right after, have 100,000 grades of relief areas that purify clean control criterions.
The back door that double door tunnel type pressure steam sterilizer of the present invention carries out discharging is right after has 100,000 grades of relief areas that purify clean control criterion (concrete standard sees the following form), the material of energy strict guarantee after the ripening sterilization can enter into strong cold house and aseptic inoculation chamber in clean control hierarchy standard after discharging like this.The temperature and pressure regulation and control of double door tunnel type pressure steam sterilizer of the present invention, all operation sequence and the control criterion of being set by computer carried out, and be highly sensitive, and chronoscope is pressed accurately, the automaticity height, and the capacity of steriliser can be set according to the scale of batch production fully.And existing double door pressure sterilization device, two-door can the unlatching simultaneously, discharge door does not have purification standard control, and capacity is little, and the energy consumption height easily pollutes.
Further again, the ripening of the culture medium of described large-scale cultivation of cordyceps militaris and sterilizing methods is characterized in that, may further comprise the steps:
A), add the water step---in described double door tunnel type pressure steam sterilizer, add the water of ripening and sterilization aequum;
B), preheating step---with added water preheat to 100 in the step a) ℃～135 ℃;
C), the product placement step---the cultivation box of the culture medium that needs ripening and sterilization will be housed, be placed on the mover after, advance in the sterile chamber of described double door tunnel type pressure steam sterilizer from the Qianmen and close the door airtight;
D), after heating steps---the cover stressor layer for the treatment of described double door tunnel type pressure steam sterilizer rises to 0.10Mpa, heated up and supercharging in the sterile chamber of described double door tunnel type pressure steam sterilizer, suitable until the cover stressor layer with sterile chamber's pressure, be stable at 0.11Mpa;
E), ripening and sterilization steps---the temperature for the treatment of the sterile chamber of described double door tunnel type pressure steam sterilizer rises to after 100 ℃～135 ℃, enter ripening and sterilization keeping warm mode, carry out ripening and sterilization, ripening and sterilization temperature retention time are 40min～50min minute;
F), steps of exhausting---get rid of the gas in the sterile chamber, after room temperature subject to sterilization is reduced to below 90 ℃ naturally, pressure reduces to zero, in the depollution of environment is controlled between 100,000 grades buffering, open the back door of described double door tunnel type pressure steam sterilizer, send mover from the back door of described double door tunnel type pressure steam sterilizer.
Compared with prior art, the ripening of the culture medium of large-scale cultivation of cordyceps militaris of the present invention and sterilizing methods have following beneficial effect:
1, the ripening of the culture medium of large-scale cultivation of cordyceps militaris of the present invention and sterilizing methods, the double door tunnel type pressure steam sterilizer of use, the more common horizontal circular steam sterilization of its unit volume output pot is high 12.5 times.Ripening can be handled and the culture medium more than five tons of sterilizing day; The common horizontal circular steam sterilization pot of same unit are originally a collection ofly can only be put 400 glass culture bottles, can criticize 250 cultivations of input boxes now, is equivalent to 5000 glass culture bottles.And every batch of ripening is doubled with the sterilization work efficiency.
2, can cooperate depollution of environment rank control requirement, guarantee product quality, be difficult for microbiological contamination;
3, automaticity is higher in the operating process;
4, thermal efficiency height in the operating process, the material ripening is thorough.Under method and apparatus condition of the present invention, quick heating, the insulated sterilizing time is long, and at 121 ℃, pressure is that temperature retention time reaches 40min under the steady-state conditions of 0.11mpa, material 100% ripening;
5, the ripening of the culture medium of large-scale cultivation of cordyceps militaris of the present invention and sterilizing methods; the double door tunnel type pressure steam sterilizer that uses; Qianmen charging, back door discharging can be realized, the working condition of ripening, sterilization and cold, inoculation working range connection by force unified clean control zone can be like this.
6, ripening and sterilization are carried out than existing a lot of enterprises ripening and sterilization being implemented step by step synchronously, have developed major step on production technology is improved.At first reduce the process of once charging and discharging, reduced labor intensity; Next has reduced the input of fixed assets and has saved production site; Its three, ripening, sterilization two steps are merged, the corresponding step that reduced has reduced energy consumption nearly 40%; Its four, in the ripening and sterilization process of every batch of culture medium, under stable high temperature, pressure and heat-retaining condition, guaranteed the ripening and the sterilization of material 100%.
The specific embodiment
Further specify technical scheme of the present invention below in conjunction with embodiment.
As the ripening and the sterilizing methods of the culture medium of large-scale cultivation of cordyceps militaris of the present invention, its maturation stage and sterilization steps merge carries out, and that is to say that the present invention unites two into one maturation stage and sterilization steps.The present invention has reached the purpose that realizes ripening simultaneously and sterilization by the temperature and time of control ripening and sterilization.
Specifically, the ripening of the culture medium of large-scale cultivation of cordyceps militaris of the present invention and sterilizing methods carry out in double door tunnel type pressure steam sterilizer.This steriliser has Qianmen and back door, and the Qianmen is the door that is used for charging, and the back door is the door that is used for discharging.Steriliser is the horizontal pressure steam sterilizer of SVW 105 types that is provided by Shanghai Huaxian Medical Nuclear Instruments Co., Ltd.; portion uses mechanical interlocking principle of the prior art within it; make it reach a unlatching; another is the result of locking voluntarily, has so just become double door tunnel type pressure steam sterilizer of the present invention.Double door tunnel type pressure steam sterilizer capacity in this specific embodiment is 0.48 cube to 3.6 cubes, has the ability of working under 0.22MPA and 135 ℃ of conditions.Double door tunnel type pressure steam sterilizer automaticity height can add water automatically.
The ripening of the culture medium of large-scale cultivation of cordyceps militaris of the present invention and sterilizing methods comprise adding water, preheating, product placement, heating, ripening and steps such as sterilization, aerofluxus.
Implement the ripening and the sterilizing methods of the culture medium of large-scale cultivation of cordyceps militaris of the present invention, at first add the water step.In this step, connect the power supply of double door tunnel type pressure steam sterilizer earlier, at this moment each meter lamp on the instrument board of double door tunnel type pressure steam sterilizer is bright entirely, button selects double door tunnel type pressure steam sterilizer to add water automatically, the buzzer of double door tunnel type pressure steam sterilizer begins buzzing, cut off the water supply after the topped up with water, the buzzer of double door tunnel type pressure steam sterilizer stops buzzing.
Add water at double door tunnel type pressure steam sterilizer and finish, buzzer stops to carry out preheating step after the buzzing.Preheating step is that operative temperature is provided with key, is 121 ℃ with thermoregulation.Double door tunnel type pressure steam sterilizer is automatically to water preheat.
Carry out the product placement step then.The product placement step is will the cultivation box of the culture medium that needs the ripening sterilization be housed, after being placed on the mover in order, advance from the Qianmen of double door tunnel type pressure steam sterilizer in the sterile chamber of double door tunnel type pressure steam sterilizer, it is airtight to close the Qianmen then.Mover is to adopt the mover of rustless steel as material manufacture.
Then carry out heating steps.Press the sterilization key on the double door tunnel type pressure steam sterilizer, setting sterilising temp is 121 ℃, ripening and sterilization temperature retention time are 45 minutes, after the cover stressor layer for the treatment of double door tunnel type pressure steam sterilizer rises to 0.10Mpa, press start key, heat up in the sterile chamber that buzzer stops double door tunnel type pressure steam sterilizer after the buzzing, sterile chamber's temperature-rise period follows the cover stressor layer to enter in the sterile chamber, this moment, the sterile chamber pressure index rose and the jacket layer pressure index reduces until last both pressure quite gradually gradually, was stable at about 0.11Mpa.
Next carry out ripening and sterilization steps.Sterile chamber's temperature for the treatment of double door tunnel type pressure steam sterilizer rises to after 121 ℃, enters ripening and sterilization keeping warm mode, and sterilizing indicator lamp is bright, and the sterilizing parameter viewing area shows the residue sterilization time.
Carry out steps of exhausting at last.After ripening and sterilization steps end, carry out steps of exhausting, get rid of the gas in the sterile chamber, make the temperature of the sterile chamber of double door tunnel type pressure steam sterilizer be reduced to below 90 ℃ naturally, after pressure reduces to zero, in being controlled between 100,000 grades buffering, the depollution of environment opens the back door of double door tunnel type pressure steam sterilizer, from the back door discharging of double door tunnel type pressure steam sterilizer.
The cultivation box of the culture medium that need ripening sterilization being housed after ripening, the sterilization is from the back door discharging of double door tunnel type pressure steam sterilizer, double door tunnel type pressure steam sterilizer back door connects the relief areas with 100,000 grades of clean control criterions of purification, 100,000 grades of relief area back that purify clean control criterion connect strong cold with 100,000 grades of clean control criterions of purification, culture medium is cooled to room temperature at strong cold, enters 10,000 grades of sterilizing room inoculations that purify clean control criterion through transmitting the storehouse again.Guaranteed like this culture matrix under the condition of the strict control of purification standard with scope in, realize finishing synchronously of ripening and sterilization, guarantee the inoculation of cordyceps species under the culture matrix aseptic condition.
Embodiment 1
Described according to the specific embodiment, 5 batches of cultivation boxes that culture medium is housed are carried out ripening of the present invention and sterilization, cultivate box for 20 every batch.After ripening and the sterilization, material 100% ripening.After these cultivation boxes that culture medium is housed are placed the cultivation cycle of a Cordyceps militaris (L.) Link., do not find that culture medium has pollution.
Those of ordinary skill in the art will be appreciated that, above embodiment is used for illustrating the present invention, and be not to be used as limitation of the invention, as long as in connotation scope of the present invention, all will drop in claims scope of the present invention variation, the modification of the above embodiment.
Claims (4)
1, a kind of ripening of culture medium of large-scale cultivation of cordyceps militaris and sterilizing methods is characterized in that: maturation stage and sterilization steps merge carries out, and the temperature of ripening and sterilization is 100 ℃～135 ℃, and the temperature retention time of ripening and sterilization is 40min～50min.
2, the ripening of the culture medium of large-scale cultivation of cordyceps militaris according to claim 1 and sterilizing methods is characterized in that: described ripening and sterilization are carried out in the double door tunnel type pressure steam sterilizer that two of front and back are arranged synchronously; The back door Qianmen double door tunnel type pressure steam sterilizer of locking voluntarily when locking, back door discharging voluntarily when described double door tunnel type pressure steam sterilizer is the Qianmen charging.
3, the ripening of the culture medium of large-scale cultivation of cordyceps militaris according to claim 2 and sterilizing methods; it is characterized in that: after ripening and sterilization, with culture medium send to that back door with described double door tunnel type pressure steam sterilizer is right after, have 100,000 grades of relief areas that purify clean control criterions.
4, the ripening of the culture medium of large-scale cultivation of cordyceps militaris according to claim 2 and sterilizing methods is characterized in that, may further comprise the steps:
A), add the water step---in described double door tunnel type pressure steam sterilizer, add the water of ripening and sterilization aequum;
B), preheating step---with added water preheat to 100 in the step a) ℃～135 ℃;
C), the product placement step---the cultivation box of the culture medium that needs ripening and sterilization will be housed, be placed on the mover after, advance in the sterile chamber of described double door tunnel type pressure steam sterilizer from the Qianmen and close the door airtight;
D), after heating steps---the cover stressor layer for the treatment of described double door tunnel type pressure steam sterilizer rises to 0.10Mpa, heated up and supercharging in the sterile chamber of described double door tunnel type pressure steam sterilizer, suitable until the cover stressor layer with sterile chamber's pressure, be stable at 0.11Mpa;
E), ripening and sterilization steps---the temperature for the treatment of the sterile chamber of described double door tunnel type pressure steam sterilizer rises to after 100 ℃～135 ℃, enter ripening and sterilization keeping warm mode, carry out ripening and sterilization, the temperature retention time of ripening and sterilization is 40min～50min minute;
F), steps of exhausting---get rid of the gas in the sterile chamber, after room temperature subject to sterilization is reduced to below 90 ℃ naturally, pressure reduces to zero, in the depollution of environment is controlled between 100,000 grades buffering, open the back door of described double door tunnel type pressure steam sterilizer, send mover from the back door of described double door tunnel type pressure steam sterilizer.
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| CN2008100416929A CN101648029B (en) | 2008-08-14 | 2008-08-14 | Method for curing and sterilizing culture media for large-scale cultivation of cordyceps militaris |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102283177A (en) * | 2011-07-22 | 2011-12-21 | 重庆市中药研究院 | Method for recycling and treating feeding matrix of aweto parasitic larva |
| CN110679395A (en) * | 2019-10-22 | 2020-01-14 | 江苏华绿生物科技股份有限公司 | Anti-pollution process in sterilization link |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN2133298Y (en) * | 1992-09-04 | 1993-05-19 | 王智铭 | Automatic control dry heating sterilizing unit with microcomputer |
| CN2763439Y (en) * | 2005-02-02 | 2006-03-08 | 李葵 | Linkage interlocking door |
| CN101177665B (en) * | 2006-11-06 | 2010-09-15 | 张刚 | Industrial method for culturing north winter worm summer herb with high-content of cordycepin |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102283177A (en) * | 2011-07-22 | 2011-12-21 | 重庆市中药研究院 | Method for recycling and treating feeding matrix of aweto parasitic larva |
| CN110679395A (en) * | 2019-10-22 | 2020-01-14 | 江苏华绿生物科技股份有限公司 | Anti-pollution process in sterilization link |
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Address after: 201203, Room 518, No. 312 blue wave road, Zhangjiang hi tech park, Shanghai, Pudong Applicant after: SHANGHAI GUOBAO ENTERPRISE DEVELOPMENT CENTER Co-applicant after: Shanghai Guobao Bioengineering Research Institute Address before: 201203, Room 518, No. 312 blue wave road, Zhangjiang hi tech park, Shanghai, Pudong Applicant before: Shanghai Guobao Enterprise Development Center Co-applicant before: Shanghai Guobao Bioengineering Research Institute |
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Effective date of registration: 20220908 Address after: No. 6, Lane 293, Fuzhou Road, Huangpu District, Shanghai, 200001 Patentee after: Tang Peichen Address before: 201203, Room 518, No. 312 blue wave road, Zhangjiang hi tech park, Shanghai, Pudong Patentee before: SHANGHAI GUOBAO ENTERPRISE DEVELOPMENT CENTER Patentee before: Shanghai Guobao Bioengineering Research Institute |