Premium
Framework for online optimization of recombinant protein expression in high‐cell‐density Escherichia coli cultures using GFP‐fusion monitoring
Author(s) -
Chae Hee Jeong,
Delisa Matthew P.,
Cha Hyung Joon,
Weigand William A.,
Rao Govind,
Bentley William E.
Publication year - 2000
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/1097-0290(20000805)69:3<275::aid-bit5>3.0.co;2-y
Subject(s) - inducer , green fluorescent protein , biological system , escherichia coli , chloramphenicol acetyltransferase , lac operon , computer science , mathematical optimization , biology , mathematics , biochemistry , gene expression , promoter , gene
A framework for the online optimization of protein induction using green fluorescent protein (GFP)‐monitoring technology was developed for high‐cell‐density cultivation of Escherichia coli. A simple and unstructured mathematical model was developed that described well the dynamics of cloned chloramphenicol acetyltransferase (CAT) production in E. coli JM105 was developed. A sequential quadratic programming (SQP) optimization algorithm was used to estimate model parameter values and to solve optimal open‐loop control problems for piecewise control of inducer feed rates that maximize productivity. The optimal inducer feeding profile for an arabinose induction system was different from that of an isopropyl‐β‐ D ‐thiogalactopyranoside (IPTG) induction system. Also, model‐based online parameter estimation and online optimization algorithms were developed to determine optimal inducer feeding rates for eventual use of a feedback signal from a GFP fluorescence probe (direct product monitoring with 95‐minute time delay). Because the numerical algorithms required minimal processing time, the potential for product‐based and model‐based online optimal control methodology can be realized. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 69: 275–285, 2000.