Application of design-of-experiments procedures to optimize efficiently pretreatment of lipase for use in a nonaqueous reaction

Rafael A. Garcia, Mark R. Riley

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

A variety of different pretreatments can improve the performance of enzymes in nonpolar reaction media. These pretreatments have primarily been studied in isolation; however, interactions between some pairs of pretreatments are known to exist. The presence of these interactions complicates the design of an optimum multifactor pretreatment. Modern design-of-experiments techniques allow the simultaneous optimization of two or more variables. To improve the performance of lipase in a model reaction, we used a technique called the method of steepest ascent to optimize three variables simultaneously: pretreatment pH and sodium phosphate concentration, and the concentration of acetic acid (one of the reactants) in the reaction mixture. In only 26 experimental runs, this optimization process determined a combination of variable settings that yielded a reaction product approx 180 times faster than achieved with untreated enzyme. Evidence is presented to demonstrate that locating this optimum with single-factor experiments would be inefficient. This article demonstrates the efficiency of the method of steepest ascent particularly for evaluation of enzymatic reaction conditions exhibiting significant interactions.

Original languageEnglish (US)
Pages (from-to)69-78
Number of pages10
JournalApplied Biochemistry and Biotechnology
Volume127
Issue number2
StatePublished - Nov 1 2005

Fingerprint

Lipases
Lipase
Design of experiments
Enzymes
Reaction products
Acetic acid
Acetic Acid
Phosphates
Sodium
Experiments
sodium phosphate

Keywords

  • Esterification
  • Fragrance
  • Lipase
  • Lyophilization
  • Nonaqueous enzymology
  • Optimization
  • Pretreatment

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Cite this

@article{cee42720cbb447f8965591aa108bc187,
title = "Application of design-of-experiments procedures to optimize efficiently pretreatment of lipase for use in a nonaqueous reaction",
abstract = "A variety of different pretreatments can improve the performance of enzymes in nonpolar reaction media. These pretreatments have primarily been studied in isolation; however, interactions between some pairs of pretreatments are known to exist. The presence of these interactions complicates the design of an optimum multifactor pretreatment. Modern design-of-experiments techniques allow the simultaneous optimization of two or more variables. To improve the performance of lipase in a model reaction, we used a technique called the method of steepest ascent to optimize three variables simultaneously: pretreatment pH and sodium phosphate concentration, and the concentration of acetic acid (one of the reactants) in the reaction mixture. In only 26 experimental runs, this optimization process determined a combination of variable settings that yielded a reaction product approx 180 times faster than achieved with untreated enzyme. Evidence is presented to demonstrate that locating this optimum with single-factor experiments would be inefficient. This article demonstrates the efficiency of the method of steepest ascent particularly for evaluation of enzymatic reaction conditions exhibiting significant interactions.",
keywords = "Esterification, Fragrance, Lipase, Lyophilization, Nonaqueous enzymology, Optimization, Pretreatment",
author = "Garcia, {Rafael A.} and Riley, {Mark R.}",
year = "2005",
month = "11",
day = "1",
language = "English (US)",
volume = "127",
pages = "69--78",
journal = "Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology",
issn = "0273-2289",
publisher = "Humana Press",
number = "2",

}

TY - JOUR

T1 - Application of design-of-experiments procedures to optimize efficiently pretreatment of lipase for use in a nonaqueous reaction

AU - Garcia, Rafael A.

AU - Riley, Mark R.

PY - 2005/11/1

Y1 - 2005/11/1

N2 - A variety of different pretreatments can improve the performance of enzymes in nonpolar reaction media. These pretreatments have primarily been studied in isolation; however, interactions between some pairs of pretreatments are known to exist. The presence of these interactions complicates the design of an optimum multifactor pretreatment. Modern design-of-experiments techniques allow the simultaneous optimization of two or more variables. To improve the performance of lipase in a model reaction, we used a technique called the method of steepest ascent to optimize three variables simultaneously: pretreatment pH and sodium phosphate concentration, and the concentration of acetic acid (one of the reactants) in the reaction mixture. In only 26 experimental runs, this optimization process determined a combination of variable settings that yielded a reaction product approx 180 times faster than achieved with untreated enzyme. Evidence is presented to demonstrate that locating this optimum with single-factor experiments would be inefficient. This article demonstrates the efficiency of the method of steepest ascent particularly for evaluation of enzymatic reaction conditions exhibiting significant interactions.

AB - A variety of different pretreatments can improve the performance of enzymes in nonpolar reaction media. These pretreatments have primarily been studied in isolation; however, interactions between some pairs of pretreatments are known to exist. The presence of these interactions complicates the design of an optimum multifactor pretreatment. Modern design-of-experiments techniques allow the simultaneous optimization of two or more variables. To improve the performance of lipase in a model reaction, we used a technique called the method of steepest ascent to optimize three variables simultaneously: pretreatment pH and sodium phosphate concentration, and the concentration of acetic acid (one of the reactants) in the reaction mixture. In only 26 experimental runs, this optimization process determined a combination of variable settings that yielded a reaction product approx 180 times faster than achieved with untreated enzyme. Evidence is presented to demonstrate that locating this optimum with single-factor experiments would be inefficient. This article demonstrates the efficiency of the method of steepest ascent particularly for evaluation of enzymatic reaction conditions exhibiting significant interactions.

KW - Esterification

KW - Fragrance

KW - Lipase

KW - Lyophilization

KW - Nonaqueous enzymology

KW - Optimization

KW - Pretreatment

UR - http://www.scopus.com/inward/record.url?scp=27644587102&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=27644587102&partnerID=8YFLogxK

M3 - Article

C2 - 16258185

AN - SCOPUS:27644587102

VL - 127

SP - 69

EP - 78

JO - Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology

JF - Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology

SN - 0273-2289

IS - 2

ER -