CN209722177U - A kind of flexible biological reactor - Google Patents

Abstract

The utility model relates to a kind of flexible biological reactors, tank body including flexible material, the tank body includes the first cavity and the second cavity to connect, first cavity is located above second cavity, first cavity is cylinder or stage body, and second cavity is vertebral body, and first cavity is equipped with inoculation mouth, culture medium entrance and gas outlet, second cavity is equipped with air inlet, and the air inlet connects air-breather.The utility model structure is simple, and suspension culture effect is good, and oxygen mass transfer coefficients are suitable for that shearing force is suitable for that cost is relatively low, is easy to be mass produced.

Description

A kind of flexible biological reactor

Technical field

The utility model relates to cell cultivation equipment field, especially a kind of flexible biological reactor.

Background technique

Bioreactor is the key equipment that scale carries out cell suspension cultures, by taking culture plant cell as an example, due to The acquiring way of the natural prodcuts of plant origin be primarily now by extracting wild plant resource or artificial growth resource, however Problem of most of wild plant resource due to excessively excavating and facing resource exhaustion.Artificial growth resource is delayed to a certain extent The natural products for having solved plant origin underproduces problem, but to be commonly present content unstable, again for the product in artificial growth source The problems such as metal is exceeded, agriculture is residual exceeded, while artificial growth easily causes the occupancy in forest land, arable land, the destruction problem of environment.Therefore Using formation for Plant Cell Suspension technology, the industrialized production of natural active product is carried out in conjunction with bioreactor technology, is to solve to plant One of the important channel of the natural products shortage of resources in object source.

Plant cell suspension culture bioreactor is sent out on the basis of traditional microbiological culture bioreactors at present Zhan Erlai, main Types have mechanical agitation type, gas-lifting type, bubble type, rotary drum and fixed bed etc..The structure of the above bioreactor Type is cylindrical structure, finds that cylinder configuration bioreactor bottom is easy deposition when plant cell suspension culture in research Compared with many cells, it is easy to appear building-up effect, causes depositional phenomenon serious especially plant cell suspension culture early period and later period cell, Growth and breeding and the secondary metabolite synthesis for directly affecting cell, cause cell-unit matrix yield and target product content low The problem of.Although agitating paddle can improve deposition to a certain extent, susceptibility ratio of the cell mass assembled to shearing force It is unicellular high more, so that a large amount of cells are dead during the culture that suspends.Gas-lifting type and Sparged bioreactors are in order to make to gather The cell mass of collection suspends, and needs sufficiently large ventilation ratio, however shearing force caused by bubbles burst while ventilation ratio increase It greatly increases, while increasing with foam, carbon dioxide is excessively escaped, and the growth and metabolism of plant cell are unfavorable for.Immobilization Culture reduces plant cell to the easy differentiating phenomenon of sensitivity and cell of shearing force to a certain extent, but natural products belongs to more In intracellular product, its popularization and application is limited.

Disposable wave bioreactor is the relatively suitable animal and plant cells suspension culture to grow up the nearly more than ten years Bioreactor, mainly by shake platform make wave effect increase gas-liquid area reach mixing and oxygen supply.But It is found in research, disposable wave bioreactor is directed to suspension culture effect easily assemble, the biggish cell mass of particle It is poor, and coefficient generally only has 50%.Chinese invention patent CN103224882A is disclosed with the primary of aerator Property wave bioreactor, but for the plant cell aggregates easily assembled, increasing aerator, the effect is unsatisfactory, Show as only having certain impetus in narrow range around the aerator, if increasing ventilatory capacity, shearing force and bubble Foam problem also increases, and is unfavorable for plant cell growth breeding, therefore bottom plane area is excessive for plant cell suspension culture It is extremely unfavorable.

Utility model content

For above situation, the utility model provides a kind of flexible biological reactor, heavy with cell in the culture that solves to suspend The problem that product problem and ventilation cause shearing force excessive.

Flexible biological reactor described in the utility model, including the integrally formed tank body of flexible material, preferred polyamide are gathered Ethylene, ethylene-vinyl acetate copolymer, ethylene-vinyl alcohol copolymer etc., for the hard material of the prior art, flexible material Tank body be easy to mass assembling, transport, irradiation, inoculated and cultured, facilitate technical scale metaplasia produce.On the other hand, cell culture Period easily adheres on tank wall, can press bag body using flexible material, adherent cell is easy to de- wall and returns to liquid In body, inoculum concentration early period can be reduced, while promoting fermentation liquid cell uniformity.

Flexible biological reactor described in the utility model forms tank body by the first cavity and the second cavity, wherein the first chamber Body undertakes main cell culture function, is also equipped with part cell in the second cavity, it is critical that by the second cavity Inclined surface design, interfered in conjunction with ventilation, solve culture plant cell in the prior art and because cell aggregation is agglomerating form precipitating The problem of plane.

Preferably, the inclination angle of second cavity at least one inclined surface is 30-70 degree, more preferably inclination angle 45-60 degree, at this time Entire cultivation cycle inner cell can be made to be in well suspended state, be conducive to cell growth and breeding, unit base cell is improved and receive Rate reduces production cost.

Preferably, using the air-distributing device, switched using aperture, the tank body will be entered by the air inlet Air-flow be dispersed into the bubble of adjustable section.

To solve this technical problem, the utility model takes following scheme:

A kind of flexible biological reactor, the tank body including flexible material, the tank body include the first cavity for connecting and Second cavity, first cavity are located above second cavity, and first cavity is cylinder or stage body, second chamber Body is vertebral body, and first cavity is equipped with inoculation mouth, culture medium entrance and gas outlet, and second cavity is equipped with air inlet Mouthful, the air inlet connects air-breather.

Further, the flexible material is polyamide, polyethylene, ethylene-vinyl acetate copolymer, ethylene-vinyl alcohol At least one of copolymer.

Further, the gas outlet is located on the first cavity top closure, and the gas outlet passes through the first pipeline Respirator is connected, the first air control valve is set on first pipeline.

Further, the air inlet is located on the inclined surface of second cavity.

Further, the inclination angle of the second cavity at least one inclined surface is 30-70 degree.

Further, the inclination angle 45-60 degree of the second cavity at least one inclined surface.

Further, the air-breather includes flowmeter, and the air inlet passes through flowmeter described in the second piping connection, The second air control valve is set on second pipeline.

Further, air-distributing device, the air-distributing device is fixedly mounted on the inclined surface of second cavity The air inlet is connected, the air-flow for entering the tank body by the air inlet is dispersed into the bubble of adjustable section.

Further, the air-distributing device includes air accommodating cabinet, and the air accommodates cabinet sealed bottom, institute The top closure for stating air accommodating cabinet is equipped with multiple equally distributed holes, and described hole at least has 2 kinds of different holes Diameter R1, R2, the cap central of the air accommodating cabinet install rotatable baffle, the side wall connection of the air accommodating cabinet The air inlet.

Further, at least one sensor reserved opening, the sensor reserved opening grafting pH is arranged in the tank wall Meter, air saturation detection device or device for detection of carbon dioxide content.

By using preceding solution, the utility model compared with prior art, is had the advantage that using flexible material Matter is convenient for scale transport and produces, and tank body includes vertebral body structure, is acted in conjunction with ventilation, realizes that preferable cell suspends and imitates Fruit.

Moreover respirator is set on the first pipeline of gas outlet connection, and play the role of the inside and outside isolation of sack, it is specific next It says, it is advantageous that: one, it prevents from surprisingly falling the extraneous miscellaneous bacteria invasion bag inner cell of sack (tank body) when pressing, causes to pollute, Jin Erfa Ferment failure.Two, miscellaneous bacteria causes in environment with being vented into environment between culture between polluting the discovery period not in time when preventing from normally cultivating Miscellaneous bacteria quantity is exceeded, and then increases the probability of cross contamination.

Further, by the different pore size in switching air-distributing device, to adapt to the dissolved oxygen amount of different cultivation stages Demand effectively reduces ventilation bring shear stress, improves production efficiency, unit base cell yield can achieve 15g/ The high density of l or more；It effectively avoids coefficient being made by the prior art because of big ventilation bring foam proliferation issues simultaneously 50% promoted to 80% or more.

Detailed description of the invention

Fig. 1 is the flexible biological structure of reactor schematic diagram that the utility model embodiment 1 provides；

Fig. 2 is the air-distributing device structural schematic diagram that the utility model embodiment 2 provides.

Specific embodiment

The technical solution of the utility model is clearly and completely retouched below in conjunction with the drawings and specific embodiments It states, it will be appreciated by those skilled in the art that following, the embodiments are a part of the embodiments of the present invention, without It is whole embodiments, is merely to illustrate the utility model, and is not construed as limitation the scope of the utility model.It is practical based on this Embodiment in novel, every other reality obtained by those of ordinary skill in the art without making creative efforts Example is applied, is fallen within the protection scope of the utility model.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer It is recommended that condition carry out.Reagents or instruments used without specified manufacturer is the routine that can be obtained by commercially available purchase Product.

It is in the description of the present invention, it should be noted that term " center ", "upper", "lower", "left", "right", " perpendicular Directly ", the orientation or positional relationship of the instructions such as "horizontal", "inner", "outside" is to be based on the orientation or positional relationship shown in the drawings, and is only For ease of description the utility model and simplify description, rather than the device or element of indication or suggestion meaning must have it is specific Orientation, be constructed and operated in a specific orientation, therefore should not be understood as limiting the present invention.In addition, term " the One ", " second ", " third " are used for descriptive purposes only and cannot be understood as indicating or suggesting relative importance.

In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " is pacified Dress ", " connected ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or integrally Connection；It can be mechanical connection, be also possible to be electrically connected；Can be directly connected, can also indirectly connected through an intermediary, It can be the connection inside two elements.For the ordinary skill in the art, above-mentioned art can be understood with concrete condition The concrete meaning of language in the present invention.

It is not the limit to flexible biological reactor described in the utility model in embodiment by taking ginseng-cell culture as an example System, flexible biological reactor described in the utility model are equally applicable for other plant cell and some animal cell cultures.

Embodiment 1:

With reference to Fig. 1, a kind of flexible biological reactor, including the first cavity 1, the second cavity 2, the first cavity 1 are located at second 2 top of cavity.First cavity 1 be cylinder, the second cavity 2 be back taper, and a side of the second cavity horizontal by 90 ° of angles, another side is horizontal by 45 ° of angles (i.e. 45 ° of inclined surface inclination angle).The base circumference of first cavity 1 and The base circumference of two cavitys 2 is overlapped, and two cavitys connect, and forms the tank body of flexible biological reactor.The material of tank body is preferred For polyamide or polyethylene or ethylene-vinyl acetate copolymer or ethylene-vinyl alcohol copolymer.

First cavity 1 is equipped with inoculation mouth 3, culture medium entrance 5 and gas outlet 4, connects piping connection.Wherein, preferably It is connected on 1 top closure of the first cavity, convenient for inoculation and charging.Control and adjustment to tank interior air pressure for convenience, Respirator 41 and the first air control valve 42 are set on the pipeline of connection gas outlet 4, so that the intracorporal gas of tank is through the first sky Gas regulating valve 42 enters respirator 41 after adjusting flow.

The inclined surface of second cavity 2 is equipped with air inlet 6, controls charge flow rate for convenience, is equipped on air inlet pipeline Flowmeter 61 and the second air control valve 62, so that filtrated air is after flowmeter 61, using the second air control valve 62 Regulation and control enter the second cavity 2.

In order to realize preferable flow perturbation effect, shear stress is minimized, while keeping cell suspended state, preferably Multiple air-distributing devices 7 are installed on the inclined surface of the second cavity 2.Air-distributing device 7 connects air inlet pipeline, will enter tank The air-flow of body is dispersed into the bubble of suitable size.Air-distributing device 7 is using the conventional distributor with porous structure.

Further, 1 side wall of the first cavity is equipped with sensor reserved opening 8, for being inserted into pH meter, air saturation detection dress It sets or device for detection of carbon dioxide content, such as dissolved oxygen meter.

The present embodiment is only exemplary, and the utility model is not limited to such structure, and those skilled in the art should think It arrives, the second cavity 2 can also be other back tapers, such as inverted pyramid, or the inverted cone with symmetry axis.Second cavity 2 inclines Inclination angle of inclined plane can also be other numerical value, such as 30 °, 40 °, 50 °, 60 ° or 70 °, inclination angle difference will affect suspended state, It can be adjusted according to the density difference of institute's Object of Development.

Embodiment 2:

The present embodiment is further improved on the basis of embodiment 1, is realized using the thinking in switching aperture to molten in tank body The adjusting of oxygen amount carries out special designing primarily directed to air-distributing device 7, referring to fig. 2.Air-distributing device 7 includes air Cabinet 71 and air hose 72 are accommodated, air hose 72 connects the side wall of air accommodating cabinet 71, and filtrated air is sent into air accommodating Inside cabinet 71.Air accommodates 71 sealed bottom of cabinet, and top closure is equipped with multiple equally distributed holes, and aperture is respectively R1, R2, wherein the aperture of aperture 73 is R1, and the aperture of macropore 74 is R2, aperture 73 and the respectively fan-shaped aggregation of macropore 74, with sky What gas accommodating cabinet 71 covered is centrally formed central symmetry, and aperture 73 be formed by it is fan-shaped be formed by with macropore 74 it is fan-shaped that This is adjacent.

Preferably, the 10um≤aperture aperture R1 < R2≤1000um is realized under suitable ventilation ratio to dissolved oxygen with meeting The adjustment of amount.It is highly preferred that 100um≤aperture the aperture R1 < R2≤500um, and aperture R1: aperture R2=1:1.5-3.5, this When suspension culture effect it is good, oxygen mass transfer coefficients be suitable for, shearing force be suitable for.

Air accommodates 71 cap central of cabinet and installs rotatable baffle 75, and baffle 75 includes multiple sectors, fan-shaped Blade is symmetrical with being centrally formed for covering, and the edge of sector and the outer of capping are concordant.It is arrived by butterfly 75 Specific position can carry out the switching between the ventilation of macropore 74 or the ventilation of aperture 75.The mode of switching control belongs to well-known technique, This will not be repeated here.

Embodiment 3:

The present embodiment is suspended by ginseng-cell for culture, describes the application method of flexible biological reactor in embodiment 2:

By ginseng-cell callus aseptic inoculation in the qualified fluid nutrient medium of sterilizing, it is placed in 22-25 DEG C, 80- Qualified loblolly shape ginseng liquid seeds are passed through the sterile access of inoculation mouth 3 by shake culture 15-25 days in 120rpm rotary shaker Pass through in the flexible biological reactor tank body of suffocating sterilization or irradiation sterilization in advance, while people through peristaltic pump that sterilizing is qualified Join cell broth to access in tank body by culture medium entrance 5, filtrated air be inputted in tank body by air inlet 6, together When filtrated air by gas outlet 4 be discharged tank body, by the second air control valve 62 adjust tank press micro-positive pressure, be placed in thermostatic chamber, 22-25 DEG C of cultivation temperature of control controls ventilation ratio in 0.12-0.24vvm by flowmeter 61.

Air-distributing device 7 is assembled into tank body before tank body sterilizing, and in 74 gears, (i.e. baffle 75 covers in aperture for adjustment 73, expose macropore 74).Packing density of the air-distributing device 7 on inclined surface is 20-60/square meter, preferably 35-45/ Square meter.Culture is ventilated early period using macropore 74, and culture density increase is gradually adjusted to aperture 73 and ventilated.According to naked eyes during culture Observe cell concentration variation, or the relative atmospheric saturation degree and titanium dioxide of the electrode detection by being inserted into sensor reserved opening 8 Carbon amounts gradually adjusts macropore 74 to 73 gear of aperture.Entire culture period suspension 20-40 days, during which sample detection cell concentration, reached Fermentation can be terminated to 15g/l or more, filter cake low-temperature vacuum drying is harvested to obtain finished product by all fermentation liquid centrifugal filtrations, is calculated Unit base cell yield is 15.78g/l.

Although the embodiments of the present invention have been shown and described above, it is to be understood that above-described embodiment is Illustratively, it should not be understood as limiting the present invention, those skilled in the art are not departing from the utility model Principle and objective in the case where above-described embodiment can be changed in the scope of the utility model, modify, replace and Modification.

Claims (10)

1. a kind of flexible biological reactor, it is characterised in that: the tank body including flexible material, the tank body include to connect One cavity and the second cavity, first cavity are located above second cavity, and first cavity is cylinder or stage body, institute Stating the second cavity is vertebral body, and first cavity is equipped with inoculation mouth, culture medium entrance and gas outlet, on second cavity Equipped with air inlet, the air inlet connects air-breather.

2. flexible biological reactor according to claim 1, it is characterised in that: the flexible material is polyamide, poly- second At least one of alkene, ethylene-vinyl acetate copolymer, ethylene-vinyl alcohol copolymer.

3. flexible biological reactor according to claim 1, it is characterised in that: the gas outlet is located at first cavity On top closure, the gas outlet passes through the first piping connection respirator, and the first air control valve is arranged on first pipeline.

4. flexible biological reactor according to claim 1, it is characterised in that: the air inlet is located at second cavity Inclined surface on.

5. flexible biological reactor according to claim 1 or 4, it is characterised in that: the second cavity at least one inclination The inclination angle in face is 30-70 degree.

6. flexible biological reactor according to claim 5, it is characterised in that: the second cavity at least one inclined surface Inclination angle 45-60 degree.

7. flexible biological reactor according to claim 1 or 4, it is characterised in that: the air-breather includes flowmeter, The air inlet passes through flowmeter described in the second piping connection, and the second air control valve is arranged on second pipeline.

8. flexible biological reactor according to claim 1 or 4, it is characterised in that: on the inclined surface of second cavity Air-distributing device is fixedly mounted, the air-distributing device connects the air inlet, will be by described in air inlet entrance The air-flow of tank body is dispersed into the bubble of adjustable section.

9. flexible biological reactor according to claim 8, it is characterised in that: the air-distributing device includes that air holds Cabinet is set, the air accommodates cabinet sealed bottom, and the top closure of the air accommodating cabinet is equipped with multiple be uniformly distributed Hole, and described hole at least has 2 kinds of different pore size R1, R2, and the cap central installation of the air accommodating cabinet can revolve The side wall of the baffle turned, the air accommodating cabinet connects the air inlet.

10. flexible biological reactor according to claim 1, it is characterised in that: at least one biography is arranged in the tank wall Sensor reserved opening, the sensor reserved opening grafting pH meter, air saturation detection device or carbon dioxide content detection dress It sets.