Equations

Enzyme bioreactors

Specific activity of enzyme Vmax = slope of [substrate] over time

V max =k x [ E] k = specific activity

k =V max /[E ]

Batch operations of stirred reactor tb = batch reaction time (h)

Km S 0 S 0−S f S0 = original substrate concentration
t b=
V max
ln
Sf
+ ( )
V max Sf = final/target concentration

Batch operations of stirred reactor with deactivation kd = deactivation constant

Km S S −S
t b=
−1
kd ( (
ln 1−k d ln 0 + 0 f
V max ,0 S f V max, 0 ))
ln 2
k d=
t half −life

Continuous operation of a stirred reactor (CSTR) D = dilution rate (h-1)

Rmax S f F = volumetric feed flow rate (l s-1) (m3.h-1)
D × ( S0 −S f ) = V = reactor volume (l) (m3)
K ❑M + Sf
D=F / V S0 = feed substrate concentration (g.l-1)
Sf = steady state substrate concentration (g.l-1)
Rmax = maximum reaction constant (gS.l-1.h-1)
Km = Michaelis constant (g.l-1)

CSTR immobilised enzyme reactor η = total effectiveness factor

v max Sf
D × ( S0 −S f ) =η
K ❑M +S f

Continuous operation of a plug flow reactor (PFR) τ = residence time (h)

Km S S −S
τ=
( V max Sf ( )
ln 0 + 0 f
V max )
τ =1/ D=V / F

PFR immobilised enzyme reactor

Km S S −S 1
τ=
( V max Sf ( )
ln 0 + 0 f
V max η )
Mass transfer effects η = total effectiveness factor
η=ηi × ηe ηi = internal effectiveness factor
ηe = external effectiveness factor
ηe equals to 1 when negligible

Thiele modulus CS,surface = concentration of substrate on surface

β=K m /C S ,surface
Sedimentation and centrifugation

Stokes equation for free settling uA = free settling velocity (m.s-1)

2
dA ×g dA = particle diameter (m)
uA= ( ρ A −ρ) g = gravitational acceleration = 9.81 (m.s-2)
18 µ
µ = solution viscosity (N.s.m−2)
ρA = particle density (kg.m-3)
ρ = solution density (kg.m-3)
1 cP = 0.001 N.s.m−2

Centrifuge equation uc = centrifuged settling velocity (m.s-1)

d A2 ( r ω2 ) dA = particle diameter (m)
u❑C = ( ρ A −ρ ) rω2 = centrifugal force (m.s-2)
18 µ
r = centrifuge radius (m)
N = centrifuge rotational speed (s-1)
ω=2 πN
ρA = particle density (kg.m-3)
ρ = solution density (kg.m-3)

Batch centrifuge equation (e.g. swinging bucket) uc = centrifuged settling velocity (m.s-1)
2
Z × g ×d A uA = free settling velocity (m.s-1)
u❑C =Z . u A = ( ρ A −ρ) Z = G-force, ratio of centrifugal acceleration, batch
18 µ
centrifuge Z factor
r = centrifuge radius (m)
r ω2
Z= ω = 2π.N
g

Bowl centrifuge Q = volumetric flowrate (ms-1.m2) (m3s-1)

Q=u A Σ Σ = sigma factor (m2)
l = length of the bowl (m)
2 πl R02 ω2 R0 = distance from axis of rotation
Σ= [ g ]
ω=2 πN

Stacked disc centrifuge Q = volumetric flowrate (ms-1.m2) (m3s-1)

Q=u A Σ uA = free settling velocity (m.s-1)
Σ = sigma factor (m2)
Σ=¿ n = number of discs
R0 = external radius of the discs (m)
R1 = internal radius of the disc (m)
g = 9.81 m.s-2
ϴ = angle of incline of the disc (degree)
Filtration and diafiltration

Normal flow filtration u = linear flowrate (m.s-1)

kΔP 1 dV k = permeability constant
u= =
µl A dt ∆P = pressure drop (Pa)
µ= viscosity (N.s.m-2)
µα ρ0 l = bed thickness (m)
A .t
( )(
V
=
2 ΔP )( VA )+( µRΔP )
M
A = filter area (m2)
V= volume (m3)
t = time (s)
t
=K 1 V + K ❑2 α = cake resistance (m.kg-1)
V ρ0 = cell suspension density (kg.m-3)
RM = filter membrane resistance (m-1.s.Pa)
µαc µR M c = concentration of biomass
K 1= 2
K ❑2=
2 A ΔP A . ΔP

Overall resistance to filtration Rc = overall resistance to filtration

R❑c =α ρ0 ( VA ) α = actual cake resistance (m.kg-1)
Ρ0 = cake density (kg.m-3)

Cake resistance α = actual cake resistance (m.kg-1)

' s
α =α ( ΔP ) α’ = specific cake resistance (m.kg-1)
∆P = pressure drop (Pa)
s = compressibility exponent
Flux equations Jw = flux rate (m.s-1)
J w = Lp ∆ P=∆ P/ µR Lp = permeability coefficient (m.s-1.Pa-1)
Rg = resistance of gel layer (may vary with time)
L p=1 /(Rg + R m) Rm = resistance of membrane (m-1.s.Pa)
µ = viscosity of permeate (N.s.m-2)
J w =Q/ A ∆P = pressure drop
Q = volumetric flow rate (m3s-1)
A = area (m2)
Cross flow filtration Q = volumetric filtration flowrate (l.h -1)
Q= j v . A=(L p . ∆ P). A A = filter area (m2)
jv = flux rate (l.m-2 .h-1)
1 ∆P = pressure drop (Pa or psi)
Rm = Lp = membrane permeability (l.m-2 .h-1. psi-1)
L❑ p
Diafiltration volume
DV =(Vol. New Buffer)/(V )=(Q . t)/V

Batch mode diafiltration D = dilution at each step

( CC )
n = step number
D−n = C = concentration of key component following diafiltration
0
(mM)
C0 = concentration of key component prior to diafiltration
n=−ln ( CC )/ln ⁡( D¿)¿
0
(mM)

C 100 – %BE
=
C0 100

Constant volume continuous diafiltration t = time for constant volume diafiltration (h)

t=− ( VQ ) ln ( CC ) 0
V = volume of solution being diafiltered (l)
Q = volumetric diafiltration flowrate (l.h-1)
C = concentration following diafiltration (mM)
 C0 = concentration prior to diafiltration (mM)
C 100 – %BE %BE = % buffer exchange in diafiltration
=
C0 100

Process chromatography

Apparent linear flowrate uapp = apparent linear flowrate or velocity (m 3 .m-2 s-1)
Q −ΔP Q = volumetric flowrate (m3 s-1)
u❑app = =B
A µl A = cross sectional area (m2)
d = diameter of column (m)
π ×d 2 −∆P = pressure drop over column (Pa)
A= µ = viscosity (N.s.m-2) N = kg. m. s-2
4
l = column length (m)
dp = particle (bead) diameter (m)
d p2 × ε 3
B= 2
ε = voidage
150 ( 1−ε )

Actual linear flowrate uactual = actual linear flowrate or velocity (m 3 .m-2 s-1)
u app Q uapp = apparent linear flowrate (m3 .m-2 s-1)
u❑actual = = ε = voidage
ε εA

Voidage V0 = void volume

V ❑0=εV ε = voidage
V = packed column volume
Total capacity DC = dynamic capacity, determined capacity
Total capacity=DC ×V V = volume

Time required until breakthrough

V
t=
Q

Residence time tres = residence time

l l = length of column
t res =
u actual uactual = actual linear flowrate

Scale up V2 = volume of large column

V 2 Dynamic capacity of large column V1 = volume of small column
=
V 1 Dynamic capacity of small column

V2
=number offold scale up
V1

l × π × d2
V =l × A=
4