CN104996303A - Efficient reactor for cultivating ginseng adventitious roots - Google Patents
Efficient reactor for cultivating ginseng adventitious roots Download PDFInfo
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- CN104996303A CN104996303A CN201510515215.1A CN201510515215A CN104996303A CN 104996303 A CN104996303 A CN 104996303A CN 201510515215 A CN201510515215 A CN 201510515215A CN 104996303 A CN104996303 A CN 104996303A
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Abstract
The invention discloses an efficient reactor for cultivating ginseng adventitious roots. The reactor comprises a first thin film air filter, a second thin film air filter, a film replaceable syringe type filter, a glass three-way valve, a threaded bottle cap, a reactor tank body, a water wave interface, a gas source body, and a double-layered glass sand gas distributor, wherein the first thin film air filter and the film replaceable syringe type filter are communicated onto the glass three-way valve through silicone tubes; the other end of the glass three-way valve is inserted into the lower part of the threaded bottle cap; the bottle mouth of the reactor tank body is connected with the threaded bottle cap through threads; the reactor tank body is connected with gas source bodies through externally buckling of the water wave interface; the tail end opening in the gas source bodies is connected with the second thin film air filter through a silicone tube; the gas source bodies are arranged in the double-layered glass sand gas distributor from the top to the bottom in parallel. The reactor is simple and convenient to operate, high in air tightness, low in time consumption and cost, is replaceable, easy to clean, high in ginseng adventitious root survival rate, good in product quality, yield in high, and short in period.
Description
Technical field
The present invention relates to a kind ofly improve ginseng adventitious root quality, novel reactor that output uses.
Background technology
Ginseng is the traditional rare traditional Chinese medicine of China, is described as the king in medicine.Its product has wide market.But the obstacle that the cycle of planting of ginseng is longer to be made the problem such as every administrative skill, soil, sick worm, residue of pesticide become ginseng of planting to be difficult to cross over.Therefore, bio-reactor pilot scale culture ginseng adventitious root is utilized to become one of effective ways solved the problem.
At present, ginseng-cell bio-reactor pilot scale culture succeeds.The two large problems of existing Application comparison bio-reactor existence widely, one, suitable for reading mostly to be smooth micro-slope uncovered for primitive organism reactor, place suitable for reading be rubbery plug lid above, bottleneck is smooth, be difficult to ensure from connections, packaging, high-temperature heat sterilization, proceed to super-clean bench, inoculate and cultivation whole process enclosed sterile state, after it airtight only depends on inoculation, with sealed membrane, preservative film, or the cluster sealing of adhesive tape etc., safety coefficient is poor, operates extensive and difficult.And in incubation, have many times, because tank body internal gas pressure is excessive, plug can be caused to skid off or come off, whole cultivation will be forced to stop.Two, primitive organism reactor bottom (source of the gas body) and tank body are one, and internal diameter is larger, make ginseng adventitious root in the training period because the effect of gas, constantly can clash into glass sand gas distributor below, excessive shearing force is produced to culture materials, suppresses the growth of ginseng adventitious root and the accumulation of active medicinal matter wherein.And because tank body is larger, it is tiny that bottommost connects silicone tube place, in the process of cleaning, sterilizing and inoculation, easily cause fragmentation or fracture, in addition, glass sand gas distributor is fine and closely woven due to pore, blocks than being easier to, its position is in again the bottommost of tank body, be difficult to process and cleaning, when blocking serious, whole tank body will lose use value.The breakage of single aspect, and whole reactor is eliminated, a kind of very large waste beyond doubt.
Summary of the invention
The object of this invention is to provide a kind of reactor of high-efficient culture ginseng adventitious root, the design of this reactor has the plurality of advantages such as easy and simple to handle, air-tightness good, save time, cost-saving, interchangeable, easy cleaning, survival rate are high, superior product quality, output are high, the cycle is short, substantially increases the Quality and yield of ginseng adventitious root from the aspect such as efficient, convenient, multiplex.
The object of the invention is to be achieved through the following technical solutions:
A reactor for high-efficient culture ginseng adventitious root, comprises the first film air filter, the second film-air filter, film changeable syringe filter disk, glass three-way, screw cap, reactor tank body, water wave interface, source of the gas body, double glazing sand gas distributor.The first film air filter and film changeable syringe filter disk are connected on glass three-way by silicone tube, the other end of glass three-way is inserted into below screw cap, the mode that bottleneck and the screw cap of reactor tank body revolve mouth with screw thread is connected, reactor tank body is connected in water wave interface exterior snap mode with source of the gas body, the end port of source of the gas body is connected with the second film-air filter by silicone tube, and be arranged in parallel in source of the gas body double glazing sand gas distributor from top to bottom.
Utilize said apparatus to cultivate a method for ginseng adventitious root, comprise the steps:
(1) by ginseng adventitious root with the form of 0.8-1.5cm section, subculture carries out the solid culture medium light culture of seedling in the mms medium containing 20-50g/L sucrose, 0.1-0.5mg/LIBA, 3-5g/Lgelrite, 5.8-6.0pH value;
(2) by subculture 18-23 days, the ginseng adventitious root being in growth animated period is directly inoculated in medium (20-50g/L sucrose with the inoculum concentration of 1-5g/L, 0.3-0.7mg/LIBA, pH value 5.8-6.0) loading amount is carry out gas-lifting type solution culture fermentation light culture in the reactor of the 65-75% of reactor cumulative volume, control temperature is 21-29 DEG C, humidity is 30-90%, and growth cycle is 6-7 week.
The present invention utilizes novel glass bio-reactor (5-20L), cultivates ginseng adventitious root, for large-scale culture ginseng adventitious root provides technical support on a small scale.Compared to prior art, tool of the present invention has the following advantages:
1, designed in the present invention reactor is suitable for reading have employed the mode that screw thread revolves mouth, inner silica gel plug is connected with groove more closely, the bottleneck of reactor and the design of bottom interface overcome a lot of shortcomings of other conventional easy reaction device, bottle cap adopts the zoarium of aluminium screw thread and silica gel plug, bottleneck inner diameter size is strict controlled in the middle and upper part of rubber stopper, add that the screw thread outside bottleneck and the screw thread on aluminium lid tightly screw, make bottle mouth position reach firm, airtight requirement.
2, to overcome when sterilizing because the bottle mouth position that is affected by thermal expansion and contraction relaxes, have tiny gap to communicate with the external world, thus be subject to the problems such as fouled by microzyme between the culture period caused.
3, decrease the various seal means that reactor is in the past taked after inoculation, enormously simplify inoculation flow process, improve safety coefficient, and save the operating time, also save production cost simultaneously.
4, reactor tank body and source of the gas body have employed the mode that water wave interface exterior snap is connected, this design easy disassembly, good leak tightness, are convenient to cleaning, can change mutually, cost-saving.In the reactor, the quality of gas distribution pattern is directly connected to the productive rate of dissolved oxygen amount, culture yeastiness, the absorption ratio of nutrient, the formation situation of secondary metabolite and end product in medium.Therefore the design of gas distributor especially need be focused on when reactor is applied.Gas distributor adopts the glass sand compacting of 0.3 ~ 0.5 μm, aperture to form, and has the characteristics such as bearing is good, heat-resisting, acid and alkali-resistance.This sparger system can be separated with tank body, and each distributor is also detachable, convenient cleaning.
5, the aluminium screw thread of reactor threaded cap and the perfection of silica gel plug are merged, well silica gel plug and bottleneck can be combined closely, and its position there will not be and skids off or obscission in all processes, being the technical once progress of bioreactor culture ginseng adventitious root, is also that ginseng adventitious root is marched toward the technical support and place mat that Chinese style and industrialization produce.
6, the novel reactor designed by the present invention adopts water wave interface mode, tank body and source of the gas body are separated, ensure that the safety of each parts significantly, be also convenient to clean, the random exchange of each parts uses also has saved cost greatly, and is easy to keeping.Use and effect of contrast primitive organism reactor are greatly improved.
7, the novel reactor designed by the present invention can promote the growth of ginseng adventitious root and the accumulation of active medicinal matter wherein well, for expanding production provides technical support and guarantee.
Accompanying drawing explanation
Fig. 1 is the structural representation of reactor of the present invention, a-film-air filter; B-film changeable syringe filter disk; C-glass three-way; D-screw cap (fusion of aluminium screw thread and silica gel plug); E-reactor tank body; F-water wave interface; G-source of the gas body; H-double glazing sand gas distributor.
Fig. 2 is the profile of screw cap;
Fig. 3 is the vertical view of screw cap, 1-threaded cap; 2-silica gel plug; 3-tri-through hole.
Embodiment
Below in conjunction with accompanying drawing, technical scheme of the present invention is further described; but be not limited thereto; everyly technical solution of the present invention modified or equivalent to replace, and not departing from the spirit and scope of technical solution of the present invention, all should be encompassed in protection scope of the present invention.
As Figure 1-3, the reactor of high-efficient culture ginseng adventitious root provided by the invention comprises the first film air filter a, second film-air filter a ', film changeable syringe filter disk b, glass three-way c, screw cap d (leaves circular port above threaded cap, it is the punching scope of threeway, silica gel plug is fixed on the inside of threaded cap, upper plane and threaded cap inner plane similar diameters), reactor tank body e, water wave interface f, source of the gas body g and double glazing sand gas distributor h, the first film air filter a and film changeable syringe filter disk b is connected on glass three-way c by silicone tube, it is longer that the other end of glass three-way c should design, below the silica gel plug that directly can be inserted into threaded cap d, to have an appointment the length of 10cm in top, avoid the condensed water produced due to ventilation in incubation to flow directly in the first film air filter a and cause polluting or blocking.Reactor tank body e is connected in water wave interface f exterior snap mode with source of the gas body g, the parallel distribution of double glazing sand gas distributor h in source of the gas body g, about 5mm of being separated by.The lower port of source of the gas body g is connected with the second film-air filter a ' by silicone tube, and the other end of the second film-air filter a ' generally adopts silicone tube or is connected on point gas port of air compressor with other specification pipelines and attaching parts.
Utilize said apparatus to cultivate the method for ginseng adventitious root, comprise the steps:
One, by ginseng adventitious root with the form of the section of about 1cm, subculture containing 3% sucrose, 0.3%IBA, 0.3%gelrite, 5.8-6.0pH value mms medium in.
Two, by subculture 18 ~ 23 days, it is carry out light culture (periodicity of illumination is 0/24h) in the reactor of the 65-75% of reactor cumulative volume that the ginseng adventitious root being in growth animated period is directly inoculated in medium loading amount.Regularly draw materials and measure adventive root fresh weight and dry weight, amount of growth, determine its growth cycle and best harvest time.
1, the temp. and humidity that optimum ginseng adventitious root grows and illuminance is determined.
By the ginseng adventitious root of squamous subculture in solid culture medium, at its vegetative stage, being placed in thermograde is cultivate under the condition of culture of 21 DEG C, 25 DEG C, 29 DEG C, finally determine that its optimal temperature is 25 ± 1 DEG C, it is cultivate under the culture environment of 30%, 60%, 90% that the adventive root getting vegetative stage is again placed in moist gradient, finally determines that optimum humidity is 60-70%.
2, the sucrose concentration in the medium of small-sized biological bioreactor culture ginseng adventitious root and IBA concentration is determined.
The ginseng adventitious root being in growth animated period cultivated under optimum environment is taken out, is placed in the triangular flask shaken cultivation of liquid nutrient medium, carries out the gradient test of sucrose concentration.Medium is based on mms, and IBA is containing 0.33%, and saccharose gradient is 2%, 3%, 5%, 7%.Finally determine that optimum sucrose concentration is 5%.Arranging IBA gradient is equally 0.3%, 0.5%, 0.7%.Finally determine that optimum IBA concentration is 0.5%.
3, the growth cycle of small-sized biological bioreactor culture ginseng adventitious root is determined.
The ginseng adventitious root growing animated period in 1 is placed in and cultivates with the 5L bio-reactor of 2 liquid nutrient mediums that are standard.Carry out the experiment determining the optimum growh cycle.Respectively at 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks results, draw growth curve.Through the repeated test determination optimum growh cycle be 6-7 week.
4, the pluses and minuses of primitive reaction device and novel reactor are compared.
(1) primitive organism reactor place suitable for reading is that rubbery plug lid is above, bottleneck is smooth, is difficult to ensure from packaging, high-temperature heat sterilization, proceed to super-clean bench, inoculation and the enclosed sterile state of cultivating in whole process, after its airtight depends on inoculation, with sealed membrane, preservative film, or the cluster sealing of adhesive tape etc., safety coefficient is poor, operating difficulties.And in incubation, have many times, and can because tank body internal gas pressure be excessive, cause plug to come off, whole cultivation stops at this point.The aluminium screw thread of novel reactor threaded cap provided by the invention and the perfection of silica gel plug are merged, thoroughly overcome the shortcoming of above-mentioned primitive reaction device, being the technical once progress of bioreactor culture ginseng adventitious root, is also that ginseng adventitious root is marched toward the technical support and place mat that Chinese style and industrialization produce.
(2) primitive organism reactor bottom and tank body are one, and internal diameter is larger, make ginseng adventitious root in the training period because the effect of gas, constantly can clash into glass sand gas distributor below, excessive shearing force is produced to culture materials, suppresses the growth of ginseng adventitious root and the accumulation of active medicinal matter wherein.And because tank body is larger, it is tiny that bottommost connects silicone tube place, in the process of cleaning, sterilizing and inoculation, easily cause fragmentation or fracture, in addition, glass sand gas distributor is fine and closely woven due to pore, blocks than being easier to, its position is in again the bottommost of tank body, be difficult to process and cleaning, when blocking serious, whole tank body will lose use value.The breakage of single aspect, and whole reactor is eliminated, a kind of very large waste beyond doubt.And the reaction gas of entirety is not easy cleaning.The design of novel reactor provided by the invention just defeats above-mentioned shortcoming one by one.The connected mode of water wave mouth is convenient, tight, adjustable, easy cleaning, cost-saving, be easy to keeping.
(3) comprehensive (1) and (2), the design of novel reactor provided by the invention has the plurality of advantages such as easy and simple to handle, air-tightness good, save time, cost-saving, interchangeable, easy cleaning, survival rate are high, superior product quality, output are high, the cycle is short, facilitate the growth of ginseng adventitious root and the accumulation of active medicinal matter wherein well, for expanding production provides technical support and guarantee.
Claims (7)
1. the reactor of a high-efficient culture ginseng adventitious root, it is characterized in that described reactor is by the first film air filter, second film-air filter, film changeable syringe filter disk, glass three-way, screw cap, reactor tank body, water wave interface, source of the gas body and double glazing sand gas distributor are formed, the first film air filter and film changeable syringe filter disk are connected on glass three-way by silicone tube, the other end of glass three-way is inserted into below screw cap, the mode that bottleneck and the screw cap of reactor tank body revolve mouth with screw thread is connected, reactor tank body is connected in water wave interface exterior snap mode with source of the gas body, the end port of source of the gas body is connected with the second film-air filter by silicone tube, be arranged in parallel in source of the gas body double glazing sand gas distributor from top to bottom.
2. the reactor of high-efficient culture ginseng adventitious root according to claim 1, is characterized in that described double glazing sand gas distributor adopts the glass sand compacting of 0.3 ~ 0.5 μm, aperture to form.
3. the reactor of high-efficient culture ginseng adventitious root according to claim 1, it is characterized in that described screw cap is made up of threaded cap and silica gel plug, silica gel plug is fixed on the inside of threaded cap.
4. utilize a method for the bioreactor culture ginseng adventitious root of the high-efficient culture ginseng adventitious root described in the arbitrary claim of claim 1-3, it is characterized in that described method step is as follows:
(1) by ginseng adventitious root with the form of 0.8-1.5cm section, subculture carries out the solid culture medium light culture of seedling in the mms medium containing 20-50g/L sucrose, 0.1-0.5mg/LIBA, 3-5g/Lgelrite, 5.8-6.0pH value;
(2) by subculture 18 ~ 23 days, the ginseng adventitious root being in growth animated period is directly inoculated in the reactor that medium loading amount is the 65-75% of reactor cumulative volume with the inoculum concentration of 1-5g/L and carries out gas-lifting type solution culture fermentation light culture, control temperature is 21-29 DEG C, humidity is 30-90%, and growth cycle is 6-7 week.
5. the method utilizing the bioreactor culture ginseng adventitious root of high-efficient culture ginseng adventitious root according to claim 4, it is characterized in that in described step (2), medium is: based on mms medium, adds 20-50g/L sucrose, 0.3-0.7mg/LIBA, pH value 5.8-6.0.
6. the method utilizing the bioreactor culture ginseng adventitious root of high-efficient culture ginseng adventitious root according to claim 4, is characterized in that, in described step (2), cultivation temperature is 25 ± 1 DEG C.
7. the method utilizing the bioreactor culture ginseng adventitious root of high-efficient culture ginseng adventitious root according to claim 4, is characterized in that, in described step (2), humidity is 60-70%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115477030A (en) * | 2022-05-18 | 2022-12-16 | 曲风采 | Split type low-oxygen quality guaranteeing and intelligent detection monitoring method for traditional Chinese medicinal materials |
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| KR100446274B1 (en) * | 2002-03-19 | 2004-09-01 | 정서구 | bio reactor possible to divide a filter |
| CN102321537A (en) * | 2011-08-31 | 2012-01-18 | 天津大学 | Bubbling bioreactor used for plant tissue culture |
| CN104509441A (en) * | 2014-12-24 | 2015-04-15 | 吉林省绿色食品工程研究院 | System for orientationally cultivating ginseng adventitious roots by dynamic balance alimentation |
| CN204394305U (en) * | 2015-01-23 | 2015-06-17 | 蔡中岳 | Thermo jug piston button cover |
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2015
- 2015-08-20 CN CN201510515215.1A patent/CN104996303A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1096540A (en) * | 1993-06-15 | 1994-12-21 | 中国科学院化工冶金研究所 | Culture of wall dependent cells using air-lift suspended micro-carrier |
| KR100446274B1 (en) * | 2002-03-19 | 2004-09-01 | 정서구 | bio reactor possible to divide a filter |
| CN102321537A (en) * | 2011-08-31 | 2012-01-18 | 天津大学 | Bubbling bioreactor used for plant tissue culture |
| CN104509441A (en) * | 2014-12-24 | 2015-04-15 | 吉林省绿色食品工程研究院 | System for orientationally cultivating ginseng adventitious roots by dynamic balance alimentation |
| CN204394305U (en) * | 2015-01-23 | 2015-06-17 | 蔡中岳 | Thermo jug piston button cover |
Non-Patent Citations (1)
| Title |
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| 由香玲等: "小型生物反应器内人参不定根的人参皂苷累积", 《西北植物学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115477030A (en) * | 2022-05-18 | 2022-12-16 | 曲风采 | Split type low-oxygen quality guaranteeing and intelligent detection monitoring method for traditional Chinese medicinal materials |
| CN115477030B (en) * | 2022-05-18 | 2023-09-01 | 吉林吉春制药股份有限公司 | Split type low-oxygen quality guarantee and intelligent detection monitoring method for traditional Chinese medicinal materials |
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Application publication date: 20151028 |