CN104328044B - A kind of illumination carbon dioxide combined regulating bioreactor - Google Patents

A kind of illumination carbon dioxide combined regulating bioreactor Download PDF

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CN104328044B
CN104328044B CN201410604418.3A CN201410604418A CN104328044B CN 104328044 B CN104328044 B CN 104328044B CN 201410604418 A CN201410604418 A CN 201410604418A CN 104328044 B CN104328044 B CN 104328044B
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carbon dioxide
light intensity
controller
culture vessel
culture
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CN201410604418.3A
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CN104328044A (en
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薛松
褚亚东
曹旭鹏
陆洪斌
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中国科学院大连化学物理研究所
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Abstract

The present invention relates to a kind of novel bioreactor, specifically in light autotrophy incubation, changed and liquid phase carbon dioxide concentration by being used for photosynthetic intensity of illumination in detecting system, calculate amount of carbon dioxide supplement speed, the dynamic regulation of carbon dioxide is carried out, the carbon dioxide supply that realization is matched with light intensity.The present invention is effectively guaranteed energy and the coordination of material inputs in photosynthesis, is particularly suited for the outdoor culture with natural light as light source.

Description

A kind of illumination-carbon dioxide combined regulating bioreactor

Technical field:

The invention belongs to a kind of novel bioreactor, the light autotrophy culture of microalgae, photosynthetic bacteria etc. is applied to.

Background technology:

Microalgae, photosynthetic bacteria etc. can utilize its photosynthetical system, efficient absorption solar energy, fixed CO2Biomass are produced, because And bio-engineering research and the concern for producing have been obtained, it is expected to the production in bioenergy, bio-based materials and high level product On be used widely.Wherein, bioreactor culture is the important step for developing such living resources application, is especially existed Need to carry out the field of large-scale culture.

In bioreactor, the energy of biological growth comes from solar energy, therefore, in scale processes, such as It is consideration and breakthrough required for bioreactor culture that what can make full use of solar energy.It is different from artificial light source, too Sunlight source has unstability so that the efficiency of light energy utilization of culture cannot be improved.Such as in theory, the efficiency of light energy utilization of microdisk electrode Close to 10%, and actually can directly be cultivated using sunshine, its efficiency of light energy utilization only has 1~2%.

In order to improve the efficiency of light energy utilization, a feasible method is by luminous energy and photosynthetic raw material --- carbon dioxide Intake match, i.e., according to luminous energy be input into dynamic regulation carbon dioxide input.At present, common carbon dioxide is dynamically adjusted Section major way is carried out according to pH value, and the time is passed through as target carbon dioxide to set pH.This regulation typically has stagnant Property and passivity afterwards.Under the conditions of natural lighting, illumination lacks regular with rain or shine changing, it is difficult under determining different illumination Optimum pH is used as control parameter.Therefore, it is necessary to find a kind of control method for being directly based upon intensity of illumination.

Sunshine is continuous spectrum, wherein only 400nm~700nm wave-length coverages are that photosynthetical system to be utilized, It is exactly so-called photosynthetically active radiation (PAR).In light autotrophy culture, whole chemical energy that biomass are accumulated are from the sun Luminous energy, and the quantity of the chemical energy that biomass are accumulated can be obtained by determining the combustion heat.Meanwhile, in normal growth process In, for specific photosynthetic organism, in specific bioreactor, optical energy utilization efficiency, the dioxy of its production biomass Changing carbon fixation efficiency etc. can be determined by independent experiment, and empirically constant is used, and this is to be estimated based on conservation of energy principle Carbon source needed for the biological growth under different illumination conditions is calculated there is provided possible.Obtaining culture systems by pH value information again can be real The liquid phase carbon dioxide concentration on border, uses mass transfercomputation formula and Henry formula, you can to obtain required gas phase titanium dioxide Concentration of carbon, use quality flowmeter gives Dynamic Matching.

When in no light stage, i.e. night, respiration can consume a certain amount of biomass, normally deposited cell is maintained In the case of work, biomass loss and unnecessary energy consumption that respiration is brought can be reduced.Such as, for microalgae, Ke Yitong Cross and determine the change of its chlorophyll fluorescence kinetics parameters and determine minimum night oxygen intake.

Based on above-mentioned analysis, the present invention is to set out to distribute effective energy and carbon source supply in bioreactor rationally, The dynamic realtime regulation and control of efficient intensity and carbon dioxide supply are realized, to improve the culture efficiency of bioreactor, reducing Energy consumption provides new technology may.

The content of the invention

It is that luminous energy is converted to biology with carbon dioxide under photosynthetical system effect for bioreactor culture essence The chemical energy of matter form storage, the present invention proposes a kind of carbon dioxide matching that light intensity can be utilized based on photosynthesis of novelty The bioreactor of regulation, on the basis of conventional bioreactor, based on energy balance, realizes energy in culture systems The coordination of amount and material inputs.The schematic diagram of reactor is shown in accompanying drawing one, specifically includes:

1st, a kind of illumination-carbon dioxide combined regulating bioreactor, it is characterised in that utilize light intensity sensor and pH Sensor detects to the incident intensity of incubator, transmitted light intensity and pH, the carbon dioxide recharge rate of computing system demand, Carry out the dynamic regulation of carbon dioxide:

(1) that 1. and 2. light intensity sensor determines is photosynthetically active radiation intensity (PAR, unit μm ol/ (m2S)), divide Incident I is not designated asinWith transmission Iout

(2) by incident intensity and culture Photosynthetic Characteristics value --- saturation light intensity, judge whether incident light reaches saturation Light intensity.When light intensity exceedes saturation light intensity, it is incident I to be pretended with saturated lightinSubsequently calculated;Saturation light intensity is determined referring to document Methods described【Chen Genyun etc., Photosynthetic and the corresponding observation procedure of carbon dioxide are inquired into,《Plant physiology is given birth to molecule Thing journal》, 2006,32 (6):691-696】

(3) using formula N=1.6~2.0 × 0.217 × K × (Iin-Iout) × A/H carries out carbon dioxide demand rate meter Calculate, its unit is g/s, and wherein (span is in 1.6~2.4x10 for H, unit J/g for biomass calorific value4J/g), biomass and two Carbonoxide fixed proportion (span 1.6~2.0), average PAR energy conversion relations (1 μm of ol/ (m2S)=0.217W/m2, With 550nm photon energys gauge), illuminating area (A, unit m2) and optical energy utilization efficiency (K, usual value 0.01~0.09);

(4) the gas concentration lwevel C (mol/L) in liquid phase is obtained to Solubilities of Carbon Dioxide curve using pH combinations pH 【Cui Qingchen etc., Marine Chemistry dictionary, Maritime Press publishes for 1993】;

(5) N, C obtained based on (3), (4), using mass transfer formula N=KL(C*- C) calculate to obtain and be expected carbon dioxide biography The carbon dioxide liquid concentration C balanced with gaseous phase partial pressure required for passing rate N*(mol/L), wherein taking mass tranfer coefficient equal to KL =0.0045m/s;

(6) the gas phase carbon dioxide concentration required for being calculated using Henry formula, P=E*C*, take Henry'S coefficient value E= 1.64x103atm;

(7) calculating needed for completing (3)-(6) in controller (4.), and according to the result of (6), adjust mass flowmenter (5., 6.), the carbon dioxide supply that acquisition is matched with light intensity;

(8) when no light, 6. 4. controller close carbon dioxide mass flowmenter, while turning down air flow meter 5. flow is the 20-50% of illumination level.

Inventive principle:

It, by biological photosynthesis, is biomass by water and carbon dioxide conversion that the essence of optical-biological reaction is, while The process of fixed luminous energy, bioreactor provides an environment for optimization for said process.Water is for bioreactor It is much excessive, therefore, it is usually required mainly for the parameter of optimization is exactly light and carbon dioxide.In order to realize that the coordination of both is matched, Need a kind of practicable real-time control means.

The present invention be considered based on energy balance effective luminous energy input and carbon dioxide mass transfer rate it Between matching, i.e. the luminous energy of real-time estimation input system can fix how many carbon dioxide, and these carbon dioxide need it is many Big mass transfer rate could be matched, final to determine to be passed through suitable carbon dioxide in gas in conjunction with water phase gas concentration lwevel Concentration.

Meanwhile, based on the consideration of night respiration effect consumption biomass, the oxygen content being passed through in gas is suitably turned down, Part respiration always is incited somebody to action, under conditions of cytoactive is maintained, it helps improve the amount of biomass, reducing energy consumption.Such as With 10 hours nights, throughput was reduced to the 20% of daytime, and the ventilation energy consumption during overall operation will drop to original three / bis-.

The invention has the advantages that:

(1) matching of energy input and carbon source input in bioreactor is realized;

(2) measured by reducing no light stage ventilation (oxygen), part suppresses microalgae respiration, reduces because breathing causes Biomass loss and extra CO2Release;

(3) optimization of throughput contributes to the reduction of vent portion energy consumption.

Brief description of the drawings:

Fig. 1, schematic device.1. incident optical sensor, 2. light transmissive sensor, 3. pH sensors, 4. controller, 5. empty Gas mass flow controller, 6. carbon dioxide mass flow controller and 7. culture vessel.

Specific embodiment

Specific apparatus structure is shown in Fig. 1:

For 500mL culture systems:500mL bubble type cylindrical reactors, bottom is the aeration board of sintered glass.Reactor 50-60cm high, diameter 5cm, wall thickness 2mm, built-in algae solution 500mL.Unilateral fluorescent lamp illumination, culture systems input light intensity 50- 2000μmol/(m2·s)。

For 100L culture systems:Transparent pane shape culture using a height of 1.2m x 0.1m x 1.1m of x long x wide is held Device, bottom is provided with gas distribution tube, and with fluorescent lamp, (culture systems input light intensity is 50-250 μm of ol/ (m2) or natural light s) (culture systems input light intensity is 50-2000 μm of ol/ (m2S)) it is light source, built-in algae solution 100L.

For above-mentioned culture systems, the outer side surface for appearing side wall in the visible ray of culture vessel is provided with the first light intensity biography 1. with the second light intensity sensor 2., 1. the first light intensity sensor is in training to sensor positioned at the top and its present position of culture vessel The top of the micro algae culturing liquid liquid level in container is supported, the second light intensity sensor is 2. positioned at the middle and lower part and its residing position of culture vessel Put the lower section of the micro algae culturing liquid liquid level in culture vessel;3. one pH sensors insert the micro algae culturing liquid in culture vessel Under liquid level;In culture vessel, 7. bottom is provided with gas distributor, and gas distributor is by pipeline respectively through MAF Controller and carbon dioxide mass flow controller are connected with air and carbon dioxide;

First light intensity sensor 1., the second light intensity sensor 2., pH sensors 3., air mass flow amount controller and dioxy Change carbon mass flow controller to be 4. connected with controller by wire respectively.4. controller is single-chip microcomputer;MAF control Device processed and carbon dioxide mass flow controller are gas mass flow gauge.

Embodiment 1,

Using Determination of Combustion Heat device to several typical microalgaes, including lsochrysis zhanjiangensis (chrysophyceae), four slit bamboo or chopped wood algae (ocean greens Algae), the combustion heat value of chlorella (green algate of fresh water) and spirulina (blue-green algae) be measured, respectively:2.2±0.08、1.7± 0.02nd, 2.0 ± 0.03 and 2.1 ± 0.01, unit x104J/g。

With lsochrysis zhanjiangensis as culture, (culture systems are defeated in the different culture systems of measure, including under fluorescent light source Enter light intensity for 50-250 μm of ol/ (m2S)) 0.5L tubular types gas lift reactor, 1L plate-type reactors, 1.5L plate-type reactors, 3L (culture systems input light intensity is 50-2000 μm of ol/ under shaking flask, 30L plate-type reactors, 100L plate-type reactors and natural light (m2S)) 100L plate-type reactors, biomass increment is respectively to the efficiency of light energy utilization that total input luminous energy is calculated during harvesting: 9.3%th, 6.0%, 6.4%, 3.9%, 3.2%, 3.5% and 1.2%.

Embodiment 2,

Under standard 3x F/2 culture medium conditions, in 500mL tubular type bioreactors, according to the ventilation of 100mL/min (illumination section adds the CO of volume ratio 2% for speed ventilation2), 14:10 Light To Dark Ratios are cultivated lsochrysis zhanjiangensis, no light section Throughput is adjusted to the 50% and 20% of normal level respectively, F when determining no light section beginning and end pointv/FmChange ratio Example, compared to normal level, 50% and 20% group declines 1.0% and 7.5% respectively, and the throughput for determining 20% is minimum ventilation Amount.Wherein low pass tolerance biomass content 0.423g/L is high compared with the 0.389g/L of normal ventilation group by about 8.7%.

Embodiment 3,

On the basis of condition of culture described in embodiment 2, maintain air be passed through speed it is constant on the basis of, use the present invention The control technique, enters Mobile state CO2The lsochrysis zhanjiangensis culture added.It is control with CMC model described in embodiment 2. Cultivation results show, using dynamically system is added, under same incubation time, and the CO in system2It is passed through total amount and drops to control Less than the 70% of group, meanwhile, biomass has the increase of 5-40% compared with control group, and the average ratio that increases is 20%.

Claims (5)

1. a kind of illumination-carbon dioxide combined regulating bioreactor, it is characterised in that:
Use side wall carries out microdisk electrode for the culture vessel (7.) of transparent material, and light source is provided with the side of culture vessel, The visible ray that light source sends is incident in culture vessel by the side transparent side wall of culture vessel, by the another of corresponding culture vessel Side transparent side wall is appeared;
The outer side surface for appearing side wall in the visible ray of culture vessel is provided with the first light intensity sensor (1.) and the second light intensity is passed Sensor (2.), the first light intensity sensor (1.) is located at the microalgae that the top of culture vessel and its present position are in culture vessel The top of nutrient solution liquid level, the middle and lower part and its present position that the second light intensity sensor (2.) is located at culture vessel is in culture appearance The lower section of the micro algae culturing liquid liquid level in device;Under micro algae culturing liquid liquid level in one pH sensors (3.) insertion culture vessel; Be provided with gas distributor in culture vessel (7.) bottom, gas distributor by pipeline respectively through air mass flow amount controller and Carbon dioxide mass flow controller is connected with air and carbon dioxide;
First light intensity sensor (1.), the second light intensity sensor (2.), pH sensors (3.), air mass flow amount controller and two Carbonoxide mass flow controller is connected by wire with controller (4.) respectively.
2. according to the reactor described in claim 1, it is characterised in that:Controller (4.) is single-chip microcomputer;MAF control Device processed and carbon dioxide mass flow controller are gas mass flow gauge.
3. the application of reactor described in a kind of claim 1 or 2, it is characterised in that:Using reactor described in claim 1 or 2 Carry out the illumination cultivation of microalgae:It is right using the first light intensity sensor (1.), the second light intensity sensor (2.) and pH sensors (3.) The incident intensity of culture vessel (7.), transmitted light intensity and pH are detected that calculating microdisk electrode system by controller (4.) needs The carbon dioxide recharge rate asked, carries out the dynamic regulation of carbon dioxide and air:
That (1) first light intensity sensor (1.), the second light intensity sensor (2.) are determined is photosynthetically active radiation intensity PAR, unit μ mol/(m2S), incident I is designated as respectivelyInWith transmission Iout
(2) by incident intensity and culture Photosynthetic Characteristics value --- saturation light intensity, judge whether incident light reaches saturation light intensity; When light intensity exceedes saturation light intensity, it is incident I to be pretended with saturated lightInSubsequently calculated;
(3) using formula N=(1.6~2.0) × 0.217 × K × (IIn-Iout) × A/H carries out carbon dioxide demand rate meter Calculate, its unit is g/s, wherein biomass calorific value H, unit J/g, and span is in (1.6~2.4) x104J/g, biomass and two Carbonoxide fixed proportion span 1.6~2.0,1 μm of ol/ (m of average PAR energy conversion relations2S)=0.217W/m2, with 550nm photon energys gauge, illuminating area A, unit m2With optical energy utilization efficiency K, K values are 0.01~0.09;
(4) the gas concentration lwevel Cmol/L in nutrient solution is obtained to Solubilities of Carbon Dioxide curve using pH combinations pH;
(5) N, C obtained based on (3), (4), using mass transfer formula N=KL (C*- C) calculate to obtain and be expected carbon dioxide transmission speed The carbon dioxide liquid concentration C balanced with gaseous phase partial pressure required for rate N*Unit mol/L, wherein taking mass tranfer coefficient equal to KL= 0.0045m/s;
(6) gas phase carbon dioxide concentration, P=E*C in the gas distributor required for being calculated using Henry formula*, take Henry'S coefficient Value E=1.64x103atm;
(7) calculating process needed for completing above-mentioned steps (3)-(6) in controller (4.), and according to the calculating knot of step (6) Really, the gas stream of air mass flow amount controller (5.) and carbon dioxide mass flow controller (6.) is adjusted by controller (4.) The gas phase carbon dioxide concentration realized in gas distributor is measured to required concentration, cultivating system is obtained the dioxy matched with light intensity Change carbon supply.
4. the application of reactor according to claim 3, it is characterised in that:
When the light culture process that microalgae is carried out using reactor described in claim 1 or 2:
When the input light intensity of microdisk electrode system is less than or equal to 10 μM of m-2s-1When, carbon dioxide quality stream is closed by controller (4.) Amount controller (6.), while turning down the flow of air flow meter (5.) to microalgae optical culture process air mass flow 20-50%.
5. the application of reactor according to claim 3, it is characterised in that:The bioreactor is applied to oneself of microalgae Support culture, including green alga, chrysophyceae, diatom or spirulina.
CN201410604418.3A 2014-10-31 2014-10-31 A kind of illumination carbon dioxide combined regulating bioreactor CN104328044B (en)

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