An efficient approach to bioconversion kinetic model generation based on automated microscale experimentation integrated with model driven experimental design

B. H. Chen, M. Micheletti, F. Baganz, John Woodley, G.J. Lye

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Reliable models of enzyme kinetics are required for the effective design of bioconversion processes. Kinetic expressions of the enzyme-catalysed reaction rate however, are frequently complex and establishing accurate values of kinetic parameters normally requires a large number of experiments. These can be both time consuming and expensive when working with the types of non-natural chiral intermediates important in pharmaceutical syntheses. This paper presents ail automated microscale approach to the rapid and cost effective generation of reliable kinetic models useful for bioconversion process design. It incorporates a model driven approach to the experimental design that minimises the number of experiments to be performed, while still generating accurate values of kinetic parameters. The approach has been illustrated with the transketolase mediated asymmetric synthesis of L-erythrulose. Experiments were performed using automated microwell studies at the 150 or 800 mu L scale. The derived kinetic parameters were then verified in a second round of experiments where model predictions showed excellent agreement with experimental data obtained under conditions not included in the original experimental design.]it comparison with conventional methodology, the modelling approach enabled a nearly 4-fold decrease in the number of experiments while the microwell experimentation enabled a 45-fold decrease in material requirements and a significant increase in experimental throughput. The approach is generic and could be applied to a wide range of enzyme catalysed bioconversions.
Original languageEnglish
JournalChemical Engineering Science
Volume64
Issue number2
Pages (from-to)403-409
ISSN0009-2509
DOIs
Publication statusPublished - 2009

Keywords

  • Transketolase
  • Kinetics
  • Automation
  • Microscale bioprocessing
  • Biochemical engineering
  • Mathematical modelling
  • Biocatalysis

Cite this

@article{d598b3e83abc4e89a23b56f1bb58b696,
title = "An efficient approach to bioconversion kinetic model generation based on automated microscale experimentation integrated with model driven experimental design",
abstract = "Reliable models of enzyme kinetics are required for the effective design of bioconversion processes. Kinetic expressions of the enzyme-catalysed reaction rate however, are frequently complex and establishing accurate values of kinetic parameters normally requires a large number of experiments. These can be both time consuming and expensive when working with the types of non-natural chiral intermediates important in pharmaceutical syntheses. This paper presents ail automated microscale approach to the rapid and cost effective generation of reliable kinetic models useful for bioconversion process design. It incorporates a model driven approach to the experimental design that minimises the number of experiments to be performed, while still generating accurate values of kinetic parameters. The approach has been illustrated with the transketolase mediated asymmetric synthesis of L-erythrulose. Experiments were performed using automated microwell studies at the 150 or 800 mu L scale. The derived kinetic parameters were then verified in a second round of experiments where model predictions showed excellent agreement with experimental data obtained under conditions not included in the original experimental design.]it comparison with conventional methodology, the modelling approach enabled a nearly 4-fold decrease in the number of experiments while the microwell experimentation enabled a 45-fold decrease in material requirements and a significant increase in experimental throughput. The approach is generic and could be applied to a wide range of enzyme catalysed bioconversions.",
keywords = "Transketolase, Kinetics, Automation, Microscale bioprocessing, Biochemical engineering, Mathematical modelling, Biocatalysis",
author = "Chen, {B. H.} and M. Micheletti and F. Baganz and John Woodley and G.J. Lye",
year = "2009",
doi = "10.1016/j.ces.2008.09.030",
language = "English",
volume = "64",
pages = "403--409",
journal = "Chemical Engineering Science",
issn = "0009-2509",
publisher = "Pergamon Press",
number = "2",

}

An efficient approach to bioconversion kinetic model generation based on automated microscale experimentation integrated with model driven experimental design. / Chen, B. H.; Micheletti, M.; Baganz, F.; Woodley, John; Lye, G.J.

In: Chemical Engineering Science, Vol. 64, No. 2, 2009, p. 403-409.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - An efficient approach to bioconversion kinetic model generation based on automated microscale experimentation integrated with model driven experimental design

AU - Chen, B. H.

AU - Micheletti, M.

AU - Baganz, F.

AU - Woodley, John

AU - Lye, G.J.

PY - 2009

Y1 - 2009

N2 - Reliable models of enzyme kinetics are required for the effective design of bioconversion processes. Kinetic expressions of the enzyme-catalysed reaction rate however, are frequently complex and establishing accurate values of kinetic parameters normally requires a large number of experiments. These can be both time consuming and expensive when working with the types of non-natural chiral intermediates important in pharmaceutical syntheses. This paper presents ail automated microscale approach to the rapid and cost effective generation of reliable kinetic models useful for bioconversion process design. It incorporates a model driven approach to the experimental design that minimises the number of experiments to be performed, while still generating accurate values of kinetic parameters. The approach has been illustrated with the transketolase mediated asymmetric synthesis of L-erythrulose. Experiments were performed using automated microwell studies at the 150 or 800 mu L scale. The derived kinetic parameters were then verified in a second round of experiments where model predictions showed excellent agreement with experimental data obtained under conditions not included in the original experimental design.]it comparison with conventional methodology, the modelling approach enabled a nearly 4-fold decrease in the number of experiments while the microwell experimentation enabled a 45-fold decrease in material requirements and a significant increase in experimental throughput. The approach is generic and could be applied to a wide range of enzyme catalysed bioconversions.

AB - Reliable models of enzyme kinetics are required for the effective design of bioconversion processes. Kinetic expressions of the enzyme-catalysed reaction rate however, are frequently complex and establishing accurate values of kinetic parameters normally requires a large number of experiments. These can be both time consuming and expensive when working with the types of non-natural chiral intermediates important in pharmaceutical syntheses. This paper presents ail automated microscale approach to the rapid and cost effective generation of reliable kinetic models useful for bioconversion process design. It incorporates a model driven approach to the experimental design that minimises the number of experiments to be performed, while still generating accurate values of kinetic parameters. The approach has been illustrated with the transketolase mediated asymmetric synthesis of L-erythrulose. Experiments were performed using automated microwell studies at the 150 or 800 mu L scale. The derived kinetic parameters were then verified in a second round of experiments where model predictions showed excellent agreement with experimental data obtained under conditions not included in the original experimental design.]it comparison with conventional methodology, the modelling approach enabled a nearly 4-fold decrease in the number of experiments while the microwell experimentation enabled a 45-fold decrease in material requirements and a significant increase in experimental throughput. The approach is generic and could be applied to a wide range of enzyme catalysed bioconversions.

KW - Transketolase

KW - Kinetics

KW - Automation

KW - Microscale bioprocessing

KW - Biochemical engineering

KW - Mathematical modelling

KW - Biocatalysis

U2 - 10.1016/j.ces.2008.09.030

DO - 10.1016/j.ces.2008.09.030

M3 - Journal article

VL - 64

SP - 403

EP - 409

JO - Chemical Engineering Science

JF - Chemical Engineering Science

SN - 0009-2509

IS - 2

ER -