JP4103288B2 - Operation control method of aeration and stirring culture tank - Google Patents
Operation control method of aeration and stirring culture tank Download PDFInfo
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- JP4103288B2 JP4103288B2 JP2000052312A JP2000052312A JP4103288B2 JP 4103288 B2 JP4103288 B2 JP 4103288B2 JP 2000052312 A JP2000052312 A JP 2000052312A JP 2000052312 A JP2000052312 A JP 2000052312A JP 4103288 B2 JP4103288 B2 JP 4103288B2
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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Description
【０００１】
【発明の属する技術分野】
本発明は溶質ガスを液中に溶解させまたは液中で発生したガスを液の外に追い出す必要のある通気攪拌槽の運転制御方法に関する。
【０００２】
【従来の技術】
攪拌槽内での化学反応や培養槽内での細胞の呼吸などにより、酸素の消費や二酸化炭素の生成が起きる槽では、最適な条件で化学反応や培養を行うために、槽内の全ての領域で酸素濃度または二酸化炭素濃度が最適な範囲に収まるように、消費した酸素の補給もしくは生成した二酸化炭素の除去が必要である。
【０００３】
従来、通気撹拌槽内の溶存酸素や溶存二酸化炭素の濃度分布を知るためにはCytotechnology,22,p87-94,1996,"Homogenisation and oxygen transfer rates in large agitated and sparged animal cell bioreactors:Some implications for growth and production"．Alvin W.Nienow etc. で論じられているように、実際の通気撹拌槽を用いて濃度分布を予想するしかなく、全く未知の通気撹拌槽内の濃度分布を定量的に予測することは不可能であった。
【０００４】
一方、攪拌槽内部の流れに関する数値解析では、kla（ガス移動容量係数）の分布まで考慮に入れた物質濃度分布の計算はされていなかった。
【０００５】
また、溶存酸素濃度を制御する方法として、特許第1552563号がある。これは溶存酸素濃度を測定するセンサーを培養槽に設置して、その指示値に合わせて培養槽の運転条件を制御する方法である。
【０００６】
【発明が解決しようとする課題】
槽内の全ての領域において、溶質ガスの濃度を最適にすることを目的とした槽の詳細構造及び運転条件の検討を行うには、槽の構造や運転条件の変化が反映されるような方法で濃度を求める必要がある。従来のような実際の槽を用いて濃度分布を測定する方法では、構造を変化させた効果を知るためには実際に槽を製作するしか方法はなかった。また、非接触で定量的に濃度を測定することはできないので、槽内分布の正確な測定は困難という問題もある。
【０００７】
また上記特許のような場合は、測定点における値しか評価できないため、槽内分布が適切かどうかは判断できない、あるいは不明であった。すなわち攪拌槽の溶質ガス濃度を制御するのに槽内の特定の点における測定値だけを直接用いる方法では、測定している点では濃度が最適であっても、他の点では溶質ガス濃度の過不足が生じている可能性がある。したがって、溶質ガス濃度を槽内の全ての領域で最適な範囲に収めるためには槽内全体の溶質ガス濃度分布を何らかの方法で求める必要がある。
【０００８】
本発明の目的は、従来は定量的に予測することが困難であった溶質ガス濃度の分布を計算評価し、その評価結果によって運転条件を決定して制御する通気攪拌槽の運転制御方法を提供することにある。
【０００９】
【課題を解決するための手段】
本発明は、通気攪拌培養槽内の溶質ガス濃度分布を最適に制御する方法であって、前記槽内の１つ以上の点で溶存酸素濃度を計測し、前記槽のガス移動容量係数（ｋｌａ）の分布を下記の式によって求め、前記分布を用いて溶存酸素濃度分布を演算し、前記計測された溶存酸素濃度を用いて前記溶存酸素濃度分布を補正し、前記補正された溶存酸素濃度分布のうち培養条件から外れている領域があるかどうかを判断し、培養条件から外れている領域がある場合は溶存酸素濃度分布があらかじめ定められた値になるまで前記培養槽の攪拌回転数、通気量、通気ガス中の酸素分圧あるいは内圧を調整することを特徴とする。
ｋｌａ = ｆ（Ｓｃ，ε，ν，α，Ｒｂ）
ただし、ｋｌａ：ガス移動容量係数（１／ｈ）、ｋｌ：液境膜移動係数（ｍ／ｓ）、 a ：単位体積当たりの気液接触面積（１／ｍ）、Ｓｃ：シュミット数 （＝ν／Ｄｇ）、ε：乱流エネルギー散逸速度（ｍ／ｓ ３ ）、ν：動粘度（ｍ ２ ／ｓ）、Ｄｇ：液相中のガスの拡散係数（ｍ ２ ／ｓ）、α：ガスホールドアップ、Ｒｂ：気泡半径（ｍ）である。
【００１２】
∂Ｃ／∂ｔ＝−ｄｉｖ（Ｃ・（Ｕｘ、Ｕｙ、Ｕｚ））＋Ｄｏ２∇２ Ｃ＋ｋｌａ（Ｃ＊−Ｃ）＋Ｇ
ただし、
Ｃ：液中の溶存酸素濃度（ｍｏｌ／ｍ３）
（Ｕｘ、Ｕｙ、Ｕｚ）：流速ベクトル（ｍ／ｓ）
Ｄｏ ２ ：液相中の酸素の拡散係数（ｍ ２ ／ｓ）
ｋｌａ：ガス移動容量係数（１／ｓ）
ｃ＊：飽和酸素濃度（ｍｏｌ／ｍ３）（＝ｐ／Ｈ）
ｐ：水圧（ａｔｍ）
Ｈ：ヘンリー定数（ａｔｍ・ｍ３／ｍｏｌ）
Ｇ：液中で単位体積・単位時間当たりに細胞が消費する溶存酸素のモル数（ｍｏｌ／ｓ・ｍ３）
また、培養中の培養槽における複数の点での溶存酸素の測定データを用いて上記の計算方法により溶存酸素濃度分布の計算を行い、その結果を利用して培養槽の溶存酸素濃度が槽内の全ての領域で培養条件を満たしているように運転制御をおこなう。
【００１３】
【発明の実施の形態】
本発明の実施例について図１、２により説明する。図１は培養槽１０の断面を表わしている。３枚羽根の２段プロペラで撹拌し、槽底に設置されたリングスパージャー通気口４から通気を行っている。１は上段のプロペラ翼を、２は下段のプロペラ翼を示し、３は攪拌機１３（図２）によって攪拌するプロペラ１、２の回転軸である。また、運転条件は、液容量１０ｍ３、撹拌回転数２５ｒｐｍ、通気量０．０３ＶＶＭである。本実施例では槽の構造及び運転条件だけから溶存酸素濃度分布を求めているので、どのような通気攪拌槽でも構造と運転条件を決定すれば、溶存酸素濃度分布を計算する事が可能である。例えばガス移動容量係数（ｋｌａ）の分布は、次式によって計算することができる。
【００１４】
kla=kl・a
kl=0.301So2 -1/2(εν)1/4
a=3α/Rb
ただし、
kl:液境膜移動係数(m/s)
So2:酸素のシュミット数 (=ν/Do2)
ε:乱流エネルギー散逸速度(m2/s3)
ν:動粘度(m2/s)
Do2:液相中の酸素の拡散係数(m2/s)
a:単位体積当たりの気液接触面積(1/m)
α:ガスホールドアップ
Rb:気泡半径(m)
上記で求めたｋｌａの分布を用いて、次の式を有限差分法により計算することで、溶存酸素濃度分布を求めることができる。
【００１５】
∂C/∂t = -div(C・(Ux,Uy,Uz))+DO2∇2C+kla(C*-C)+G
ただし、
C :液中の溶存酸素濃度(mol/m3)
(Ux,Uy,Uz) :流速ベクトル(m/s)
kla :ガス移動容量係数(1/s)
C* :飽和酸素濃度(mol/m3) (=p/H)
p :水圧(atm)
H :ヘンリー定数(atm・m3/mol)
G :液中で単位体積・単位時間当たりに細胞が消費する溶存酸素のモル数(mol/s・m3)
次に酸素濃度分布の計算を利用して制御を行う培養槽の運転制御の実施例について述べる。通気培養槽に取り付けた１つ以上の溶存酸素計１２で溶存酸素を測定し信号を電子計算機１６に送る。電子計算機１６では、まず通気攪拌槽の構造及び運転条件を境界条件及び方程式の係数として流れの数値解析をした結果からkla分布を求め、そのkla分布を用いて溶存酸素濃度分布を計算する。その後、溶存酸素計１２から信号として送られた溶存酸素濃度で補正を行う。溶存酸素濃度の値が培養条件からはずれた領域が存在した場合、電子計算機は攪拌回転数、通気量、通気ガス中の酸素分圧、及び内圧のいずれかもしくは全部を変更して溶存酸素濃度分布を計算し直す。溶存酸素の最大値及び最小値が培養条件に適合する値が得られるまでパラメータの変更及び計算を繰り返した後、攪拌機１３、酸素分離器１５、圧力調整器１４を操作して計算で求めた攪拌回転数、通気量、酸素分圧及び内圧に実際の培養槽の運転条件を変更する。この操作を続けることで溶存酸素濃度分布が最適な運転条件で培養を行うことができる。
【００１６】
計算機１６におけるこれらの処理フローの例を図3に示す。ステップ３０では溶存酸素計１２からの信号を取り込む。ステップ３２ではガス移動容量係数ｋｌａを計算し、そしてステップ３０で取り込んだ溶存酸素により補正する。この演算された溶存酸素濃度分布の値が培養条件から外れた領域があるかどうかをステップ３４で判定し、外れている領域がある場合は、ステップ３６で攪拌回転数、通気量、通気ガス中の酸素分圧、および内圧のいずれか、または全部を変更して溶存酸素濃度分布を計算し直す。パラメータ変更計算を繰り返したあとで、実際の培養槽の運転条件をステップ３８で変更する。ステップ４０では溶存酸素が所定の範囲内にあるかどうかを判断し、満足していれば処理を終了する。満足していない場合はステップ３２以降の処理を繰り返す。このように予め定められた範囲に入るように制御することによって、培養成生物の品質の向上にもつながる。
【００１７】
また、溶存濃度が所定の範囲内にあるようにするガスは、酸素に限定されるものではなく、通気撹拌培養槽内で消費され、または生成される結果として、気泡から液中に溶け込むあるいは液中から気泡内に追い出される溶質ガスであればどのようなものでも適用できる。具体的には図３と同様な計算を、高濃度になった場合は菌の活動を阻害する二酸化炭素(菌の活動に伴い菌から液中に放出される)について行うことで、溶存二酸化炭素濃度分布を求めること及び溶存二酸化炭素濃度を無害な濃度に保つよう培養槽を制御することも可能である。
【００１８】
物質の保存則を表す微分方程式を離散化して溶質ガス濃度分布を数値解析する方法であってもよい。例えば、(溶質ガス濃度の時間変化)=(流れによる溶質ガスの流入流出量)+(拡散による溶質ガスの流入流出量)+(気液間のガス移動量)+(溶質ガスが微生物や細胞の活動又は化学反応により消費又は生成される量)、関係を利用すればよい。
【００１９】
【発明の効果】
本発明によれば、従来不可能であった通気撹拌槽内の溶質ガス濃度分布を構造及び運転条件だけから計算し、最適な値に制御することができる。したがって、槽構造の変更や新規設計を行った場合に、実際の槽を製作し溶質ガス濃度分布を測定することなく計算のみで予測することが出来る。
【００２０】
また、本発明によれば通気撹拌槽内の全ての点における溶質ガス濃度の最大値及び最小値が培養条件に収まるように、運転条件を制御することが可能であるため、培養生産物の品質維持に効果がある。
【図面の簡単な説明】
【図１】本発明の一実施例の数値解析による２段プロペラ翼を使用した通気撹拌槽の断面と溶存酸素濃度分布を示す図である。
【図２】本発明による通気撹拌培養槽の運転制御の一実施例を示すブロック構成図である。
【図３】溶存酸素濃度分布計算を制御用電子計算機で行なわせる場合の処理フロー図の一例を示す図である。
【符号の説明】
1…上プロペラ翼、2…下プロペラ翼、３…軸、４…リングスパージャー通気口、1０…培養槽、１2…溶存酸素計(一つ以上)、１３…攪拌機、１４…圧力調整器、１５…酸素分離器、１６…制御用電子計算機[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an operation control method for an aeration and stirring tank in which a solute gas needs to be dissolved in a liquid or a gas generated in the liquid needs to be driven out of the liquid.
[0002]
[Prior art]
In a tank where oxygen consumption or carbon dioxide production occurs due to chemical reaction in the stirring tank or cell respiration in the culture tank, all chemicals in the tank must be It is necessary to replenish the consumed oxygen or remove the produced carbon dioxide so that the oxygen concentration or carbon dioxide concentration falls within the optimum range in the region.
[0003]
Conventionally, Cytotechnology, 22, p87-94, 1996, “Homogenisation and oxygen transfer rates in large agitated and sparged animal cell bioreactors: Some implications for growth and production ". As discussed in Alvin W. Nienow etc., it is only possible to predict the concentration distribution using an actual aeration and stirring tank, and it is impossible to quantitatively predict the concentration distribution in a completely unknown aeration and stirring tank. Met.
[0004]
On the other hand, in the numerical analysis on the flow inside the stirring tank, the calculation of the substance concentration distribution taking into account the distribution of kla (gas transfer capacity coefficient) has not been performed.
[0005]
Japanese Patent No. 1555663 is a method for controlling the dissolved oxygen concentration. This is a method in which a sensor for measuring the dissolved oxygen concentration is installed in a culture tank, and the operating conditions of the culture tank are controlled in accordance with the indicated value.
[0006]
[Problems to be solved by the invention]
To examine the detailed structure and operating conditions of the tank for the purpose of optimizing the concentration of the solute gas in all areas in the tank, a method that reflects changes in the tank structure and operating conditions It is necessary to obtain the concentration at In the conventional method of measuring the concentration distribution using an actual tank, the only way to know the effect of changing the structure is to actually manufacture the tank. In addition, since the concentration cannot be measured quantitatively in a non-contact manner, there is a problem that accurate measurement of the distribution in the tank is difficult.
[0007]
In the case of the above-mentioned patent, only the value at the measurement point can be evaluated, so it is impossible to determine whether the distribution in the tank is appropriate or unknown. That is, in the method of directly using only the measured value at a specific point in the tank to control the solute gas concentration in the stirring tank, even if the concentration is optimal at the measurement point, the solute gas concentration at other points is controlled. There may be excess or deficiency. Therefore, in order to keep the solute gas concentration within the optimum range in all regions in the tank, it is necessary to obtain the solute gas concentration distribution in the entire tank by some method.
[0008]
An object of the present invention is to provide an operation control method for an aeration and agitation tank that calculates and evaluates the distribution of solute gas concentration, which has been difficult to predict quantitatively in the past, and determines and controls operation conditions based on the evaluation results There is to do.
[0009]
[Means for Solving the Problems]
The present invention is a method for optimally controlling the solute gas concentration distribution in an aeration and agitation culture tank, wherein the dissolved oxygen concentration is measured at one or more points in the tank, and the gas transfer capacity coefficient (kl) of the tank is measured. ) Is calculated by the following formula, the dissolved oxygen concentration distribution is calculated using the distribution, the dissolved oxygen concentration distribution is corrected using the measured dissolved oxygen concentration, and the corrected dissolved oxygen concentration distribution is corrected. And if there is a region that is out of the culture conditions, and if there is a region that is out of the culture conditions, the agitation speed and aeration of the culture tank until the dissolved oxygen concentration distribution reaches a predetermined value. It is characterized by adjusting the amount, oxygen partial pressure or internal pressure in the aeration gas.
kla = f (Sc, ε, ν, α, Rb)
Kl: gas transfer capacity coefficient (1 / h), kl: liquid film transfer coefficient (m / s), a : gas-liquid contact area per unit volume (1 / m), Sc: Schmitt number (= Ν / Dg), ε: Turbulent energy dissipation rate (m / s 3 ), ν: Kinematic viscosity (m 2 / s), Dg: Diffusion coefficient of gas in liquid phase (m 2 / s), α : Gas hold-up, Rb: Bubble radius (m).
[0012]
∂C / ∂t = -div (C · (Ux, Uy, Uz)) + Do 2 ∇ 2 C + kla (C * -C) + G
However,
C: Dissolved oxygen concentration in the liquid (mol / m 3 )
(Ux, Uy, Uz): Flow velocity vector (m / s)
Do 2 : Diffusion coefficient of oxygen in liquid phase (m 2 / s)
kl: Gas transfer capacity coefficient (1 / s)
c *: saturated oxygen concentration (mol / m 3 ) (= p / H)
p: Water pressure (atm)
H: Henry's constant (atm · m 3 / mol)
G: Number of moles of dissolved oxygen consumed by cells per unit volume / unit time in liquid (mol / s · m 3 )
In addition, the dissolved oxygen concentration distribution is calculated by the above calculation method using the measured data of dissolved oxygen at multiple points in the culture tank during the culture, and the dissolved oxygen concentration in the culture tank is calculated using the results. Control the operation so that the culture conditions are satisfied in all areas.
[0013]
DETAILED DESCRIPTION OF THE INVENTION
An embodiment of the present invention will be described with reference to FIGS. FIG. 1 shows a cross section of the
[0014]
kla = kl ・ a
kl = 0.301S o2 -1/2 (εν) 1/4
a = 3α / Rb
However,
kl: Liquid film transfer coefficient (m / s)
S o2 : Oxygen Schmidt number (= ν / D o2 )
ε: Turbulent energy dissipation rate (m 2 / s 3 )
ν: Kinematic viscosity (m 2 / s)
D o2 : Oxygen diffusion coefficient in liquid phase (m 2 / s)
a: Gas-liquid contact area per unit volume (1 / m)
α: Gas hold up
Rb: Bubble radius (m)
The dissolved oxygen concentration distribution can be obtained by calculating the following equation by the finite difference method using the distribution of kl obtained above.
[0015]
∂C / ∂t = -div (C ・ (Ux, Uy, Uz)) + D O2 ∇ 2 C + kla (C * -C) + G
However,
C: Dissolved oxygen concentration in the liquid (mol / m 3 )
(Ux, Uy, Uz): Velocity vector (m / s)
kla: Gas transfer capacity coefficient (1 / s)
C * : Saturated oxygen concentration (mol / m 3 ) (= p / H)
p: Water pressure (atm)
H: Henry's constant (atm ・ m 3 / mol)
G: mol number of dissolved oxygen consumed by cells per unit volume / unit time in liquid (mol / s ・ m 3 )
Next, an embodiment of operation control of a culture tank that performs control using calculation of oxygen concentration distribution will be described. The dissolved oxygen is measured by one or more dissolved
[0016]
An example of these processing flows in the
[0017]
Further, the gas that causes the dissolved concentration to be within a predetermined range is not limited to oxygen, but is dissolved or generated in the liquid from bubbles as a result of being consumed or generated in the aeration and agitation culture tank. Any solute gas that can be expelled from the inside into bubbles can be applied. Specifically, the same calculation as in FIG. 3 is performed on carbon dioxide that inhibits the activity of bacteria when the concentration becomes high (dissolved carbon dioxide is released from the bacteria into the liquid with the activity of the bacteria), so that dissolved carbon dioxide It is also possible to determine the concentration distribution and to control the culture tank so that the dissolved carbon dioxide concentration is kept harmless.
[0018]
A method of numerically analyzing the solute gas concentration distribution by discretizing a differential equation representing a conservation law of a substance may be used. For example, (Change in solute gas concentration with time) = (Inflow / outflow amount of solute gas due to flow) + (Inflow / outflow amount of solute gas due to diffusion) + (Amount of gas transfer between gas and liquid) + (solute gas is microbial or The amount consumed or generated by the activity or chemical reaction), and the relationship.
[0019]
【The invention's effect】
According to the present invention, it is possible to calculate the solute gas concentration distribution in the aeration and stirring tank, which has been impossible in the past, only from the structure and operating conditions, and to control the distribution to an optimum value. Therefore, when the tank structure is changed or a new design is made, it can be predicted by calculation alone without manufacturing an actual tank and measuring the solute gas concentration distribution.
[0020]
In addition, according to the present invention, the operating conditions can be controlled so that the maximum value and the minimum value of the solute gas concentration at all points in the aeration and stirring tank are within the culture conditions. It is effective for maintenance.
[Brief description of the drawings]
FIG. 1 is a diagram showing a cross section of a vented stirring tank using a two-stage propeller blade and a dissolved oxygen concentration distribution by numerical analysis of one embodiment of the present invention.
FIG. 2 is a block diagram showing an embodiment of operation control of an aeration and agitation culture tank according to the present invention.
FIG. 3 is a diagram showing an example of a processing flow chart when a dissolved oxygen concentration distribution calculation is performed by a control computer.
[Explanation of symbols]
1 ... Upper propeller blade, 2 ... Lower propeller blade, 3 ... Shaft, 4 ... Ring sparger vent, 10 ... Culture tank, 12 ... Dissolved oxygen meter (one or more), 13 ... Stirrer, 14 ... Pressure regulator, 15 ... oxygen separator, 16 ... control computer
Claims (1)
ｋｌａ = ｆ（Ｓｃ，ε，ν，α，Ｒｂ）
ただし、ｋｌａ：ガス移動容量係数（１／ｈ）、ｋｌ：液境膜移動係数（ｍ／ｓ）、 a ：単位体積当たりの気液接触面積（１／ｍ）、Ｓｃ：シュミット数 （＝ν／Ｄｇ）、ε：乱流エネルギー散逸速度（ｍ／ｓ ３ ）、ν：動粘度（ｍ ２ ／ｓ）、Ｄｇ：液相中のガスの拡散係数（ｍ ２ ／ｓ）、α：ガスホールドアップ、Ｒｂ：気泡半径（ｍ）である。 In the method for optimally controlling the solute gas concentration distribution in the aeration and agitation culture tank, the dissolved oxygen concentration is measured at one or more points in the tank, and the distribution of gas transfer capacity coefficient (kl) of the tank is The calculated dissolved oxygen concentration distribution is calculated using the distribution, the dissolved oxygen concentration distribution is corrected using the measured dissolved oxygen concentration, and the corrected dissolved oxygen concentration distribution is out of the culture condition. If there is a region that is out of the culture conditions, the stirring vessel rotation speed, the aeration rate, and the aeration gas until the dissolved oxygen concentration distribution reaches a predetermined value. An operation control method for an aeration and agitation culture tank characterized by adjusting an oxygen partial pressure or an internal pressure.
kla = f (Sc, ε, ν, α, Rb)
Kl: gas transfer capacity coefficient (1 / h), kl: liquid film transfer coefficient (m / s), a : gas-liquid contact area per unit volume (1 / m), Sc: Schmitt number (= Ν / Dg), ε: Turbulent energy dissipation rate (m / s 3 ), ν: Kinematic viscosity (m 2 / s), Dg: Diffusion coefficient of gas in liquid phase (m 2 / s), α : Gas hold-up, Rb: Bubble radius (m).
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| US7771988B2 (en) | 2005-03-24 | 2010-08-10 | Hitachi, Ltd. | Control device for fermenter |
| JP5151932B2 (en) * | 2008-11-26 | 2013-02-27 | 株式会社Ｉｈｉ | Measuring apparatus and method, and operation apparatus and method of culture tank system |
| JP6924653B2 (en) * | 2017-08-30 | 2021-08-25 | 株式会社日立プラントサービス | Mixing device and mixing method |
| JP7006361B2 (en) * | 2018-02-21 | 2022-01-24 | 住友金属鉱山株式会社 | Calculation method of gas-liquid boundary area and position design method of gas inlet |
| JP7459244B2 (en) * | 2019-10-09 | 2024-04-01 | ベーリンガー インゲルハイム インターナショナル ゲゼルシャフト ミット ベシュレンクテル ハフツング | Bioreactors or fermentors for the cultivation of cells or microorganisms in suspension on an industrial scale |
| CN115496281A (en) * | 2022-09-21 | 2022-12-20 | 中国农业大学 | Method for predicting distribution of dissolved oxygen in circulating water aniseed culture pond |
| WO2025229841A1 (en) * | 2024-05-02 | 2025-11-06 | 千代田化工建設株式会社 | Calculation model generation method, calculation method, calculation model, calculation model generation program, and calculation model generation device |
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