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Purification of enzymes by liquid-liquid extraction

  • Maria-Regina Kula
  • Karl Heinz Kroner
  • Helmut Hustedt
Conference paper
Part of the Advances in Biochemical Engineering book series (ABE, volume 24)

Abstract

The article reviews the current status of the application of aqueous two-phase systems for the extractive purification of enzymes, especially with regard to large-scale processing. The method can be used for the separation of proteins from cell debris as well as for further purification. The latter can be performed by a series of single step partitions, and apparently also by continuous multistage processes. The specificity and selectivity of extraction can be enhanced by introducing specific or general ligands. Scale-up of extractive enzyme purification is relatively simple utilizing commercially available equipment and machinery common in the chemical industry. Besides the technical performance, economic considerations also indicate the feasibility of the method at production scale.

Keywords

Partition Coefficient Polyethylene Glycol Formate Dehydrogenase Bottom Phase Binodal Curve 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Symbols

CT

concentration in the top phase (U l−1, kg l−1, mol l −1)

CB

concentration in the bottom phase (U l−1, kg l−1, mol l−1)

Ce

emergent concentration (U l−1, kg ;−1, mol l−1)

Ci

initial concentration (U l−1, kg l−1, mol l−1)

D

diameter of droplet or particle (m)

Dlim

limit droplet, (particle) diameter (m)

E

extraction factor

F

Faraday constant (96,485 C mol−1)

F*

clarifying surface (m2)

g

earth acceleration (m s−1)

G

partition ratio

k

Boltzmann constant (1.381×10−23 JK−1)

K

partition coefficient

Ki

inhibition constant (mol l−1)

Km

Michaelis Menten constant (mol l−1)

Kp

partition coefficient of protein

Kp0

partition coefficient of protein when phase potential or charge of protein is zero

K

partition coefficient of small anion

K+

partition coefficient of small cation

KE

partition coefficient of enzyme

KF

partition coefficient of polyethylene glycol dye-derivative

M

molecular weight (g mol−1)

Mw

weight average molecular weight (g mol−1)

Mn

number average molecular weight (g mol−1)

Mr

relative molecular weight

N

Newton (kg m s−2)

mN

milli Newton (10−3 kg m s−2)

N*

number of theoretical stages

n

number of binding sites

mPa

milli Pascal (10−3 N m−2)

PT

purity of top phase (%)

PB

purity of bottom phase (%)

QB

volumetric flow of bottom phase (l h−1)

QF

volumetric flow of feed (l h−1)

Qlim

limiting flow capacity (l h−1)

Qp

volumetric flow through the nozzles (l h−1)

QT

volumetric flow of top phase (l h−1)

rs

radius of interphase position (m)

ru

radius of upper phase outlet (m)

rl

radius of lower phase outlet (m)

R

general gas constant (8.314 J mol−1 K−1)

Se

separation efficiency

T

absolute temperature (K)

U

unit of enzyme activity (µMol min−1)

VT

volume of top phase (l)

VB

volume of bottom phase (l)

V′f

volume ratio of overflow and underflow

Vf

volume, ratio of upper and lower phase

Vg

sedimentation velocity (m s−1)

YT

yield of enzyme in top phase (%)

YB

yield of enzyme in the bottom phase (%)

Zp

number of charges of protein

Z+

number of charges of cation

Z

number of charges of anion

α

angle (degree)

ϱu

density of upper phase (kg m−3)

ϱ1

density of lighter phase (kg m−3)

Δϱ

density difference (kg m−3)

λ

proportionality factor of interacting forces (J g mol−1)

Σ

Sigma factor, for comparism of centrifuges (m2)

ψ

interphase potential (mV)

ω

angular velocity (rad s−1)

η

dynamic viscosity (kg s−1 m−1)

[η]

limiting viscosity (kg s−1 m−1)

τ

residence time (s)

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Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • Maria-Regina Kula
    • 1
  • Karl Heinz Kroner
    • 1
  • Helmut Hustedt
    • 1
  1. 1.Gesellschaft für Biotechnologische Forschung mbHBraunschweig-StöckheimFRG

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