Enzyme Stabilization and Immobilization: Methods and Protocols

231 Pages·2017·6.677 MB·English

by  Shelley D. Minteer (eds.)

Methods in
Molecular Biology 1504
Shelley D. Minteer Editor
Enzyme
Stabilization and
Immobilization
Methods and Protocols
Second Edition
M M B
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life and Medical Sciences
University of Hertfordshire
Hatfield, Hertfordshire , AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651
Enzyme Stabilization
and Immobilization
Methods and Protocols
Second Edition
Edited by
Shelley D. Minteer
Departments of Chemistry and Department of Materials Science and Engineering,
University of Utah, Salt Lake City, UT, USA
Editor
Shelley D. Minteer
Departments of Chemistry and Materials Science
and Engineering
University of Utah
Salt Lake City, UT, USA
ISSN 1064-3745 ISSN 1940-6029 (electronic)
Methods in Molecular Biology
ISBN 978-1-4939-6497-0 ISBN 978-1-4939-6499-4 (eBook)
DOI 10.1007/978-1-4939-6499-4
Library of Congress Control Number: 2016951931
© Springer Science+Business Media New York 2 017
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Prefa ce
Enzyme stabilization has been an area of research interest since the 1950s, but in the last
three decades, researchers have made tremendous progress in the fi eld. This has opened up
new opportunities for enzymes in molecular biology as well as industrial applications, such
as bioprocessing, biosensors , and biofuel cells .
The fi rst chapter introduces the reader to the fi eld of enzyme stabilization and the dif-
ferent theories of enzyme stabilization, including the use of immobilization as a stabiliza-
tion technique. The fi rst part of the book will focus on protocols for enzyme stabilization
in solutions including liposome formation, m icelle introduction, crosslinking, and addi-
tives, while the second part of the book will focus on protocols for enzyme stabilization
during enzyme immobilization including common techniques like sol-gel encapsulation ,
polymer encapsulation, and covalent attachment to supports. Protocols for a variety of
enzymes are shown, but the enzymes are chosen as examples to show that these protocols
can be used for both enzymes of biological importance and enzymes of industrial impor-
tance. The fi nal chapter will detail spectroscopic protocols, methods, and assays for study-
ing the effectiveness of the enzyme stabilization and immobilization strategies.
The chapters of this volume should provide molecular biologists, biochemists, and bio-
medical and biochemical engineers with the state-of-the-art technical information required
to effectively stabilize their enzyme of interest in a variety of environments (i.e., harsh tem-
perature, pH, or s olvent conditions).
Salt Lake City, UT, USA Shelley D. Minteer
v
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i x
1 I ntroduction to the Field of Enzyme Immobilization and Stabilization . . . . . . 1
Michael J. Moehlenbrock and Shelley D. Minteer
2 S tabilization of Enzymes Through Encapsulation in Liposomes. . . . . . . . . . . . 9
Makoto Yoshimoto
3 M icellar Enzymology for Thermal, pH, and Solvent Stability. . . . . . . . . . . . . . 1 9
Shelley D. Minteer
4 L ipase Activation and Stabilization in Room-Temperature Ionic Liquids . . . . . 25
Joel L. Kaar
5 N anoporous Gold for Enzyme Immobilization . . . . . . . . . . . . . . . . . . . . . . . . 3 7
Keith J. Stine , K enise Jefferson , and Olga V. Shulga
6 E nzyme Stabilization via Bio-Templated Silicification Reactions. . . . . . . . . . . . 61
Glenn R. Johnson and Heather R. Luckarift
7 C ovalent Immobilization of Enzymes on Eupergit® Supports: Effect
of the Immobilization Protocol. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Zorica D. Knežević-Jugović , S anja Ž. Grbavćić , J elena R. Jovanović ,
Andrea B. Stefanović , D ejan I. Bezbradica , D ušan Ž. Mijin ,
and M irjana G. Antov
8 M icellar Polymer Encapsulation of Enzymes . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3
Sabina Besic and Shelley D. Minteer
9 C ross-Linked Enzyme Aggregates for Applications in Aqueous
and Nonaqueous Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 09
Ipsita Roy , J oyeeta Mukherjee , and Munishwar N. Gupta
10 P rotein-Coated Microcrystals, Combi-Protein-Coated Microcrystals,
and Cross-Linked Protein-Coated Microcrystals of Enzymes for Use
in Low-Water Media. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
Joyeeta Mukherjee and Munishwar N. Gupta
11 M acroporous Poly(GMA-co-EGDMA) for Enzyme Stabilization. . . . . . . . . . . 1 39
Nenad B. Milosavić , R adivoje M. Prodanović , D ušan Velićković ,
and A leksandra Dimitrijević
12 C ytochrome c Stabilization and Immobilization in Aerogels . . . . . . . . . . . . . . 1 49
Amanda S. Harper-Leatherman , J ean Marie Wallace ,
and D ebra R. Rolison
13 E nzyme Immobilization and Mediation with Osmium Redox Polymers. . . . . . 1 65
Gaige R. VandeZande , J asmine M. Olvany , J ulia L. Rutherford ,
and M ichelle Rasmussen
vii
viii Contents
14 F errocene-Modified Linear Poly(ethylenimine) for Enzymatic
Immobilization and Electron Mediation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181
David P. Hickey
15 F AD-Dependent Glucose Dehydrogenase Immobilization
and Mediation Within a Naphthoquinone Redox Polymer. . . . . . . . . . . . . . . . 193
Ross D. Milton
16 L ayer-by-Layer Assembly of Glucose Oxidase on Carbon Nanotube
Modified Electrodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203
Alice H. Suroviec
17 K inetic Measurements for Enzyme Immobilization . . . . . . . . . . . . . . . . . . . . . 2 15
Michael J. Cooney
Index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
Contributors
MIRJANA G. ANTOV • Faculty of Technology, U niversity of Novi Sad , N ovi Sad, S erbia
SABINA BESIC • Department of Chemistry , S aint Louis University , S t. Louis, M O, U SA
DEJAN I. BEZBRADICA • Department of Biochemical Engineering and Biotechnology,
Faculty of Technology and Metallurgy , U niversity of Belgrade , B elgrade, S erbia
MICHAEL J. COONEY • Hawaii Natural Energy Institute, School of Ocean and Earth
Science and Technology, U niversity of Hawaii-M anoa , H onolulu, H I, U SA
ALEKSANDRA DIMITRIJEVIĆ • Department of Biochemistry, Faculty of Chemistry, University
of Belgrade , B elgrade, S erbia
SANJA Ž. GRBAVČIĆ • Faculty of Technology and Metallurgy, Innovation Centre,
University of Belgrade , B elgrade, S erbia
MUNISHWAR N. GUPTA • Department of Biochemical Engineering and Biotechnology,
Indian Institute of Technology Delhi , N ew Delhi, I ndia
AMANDA S. HARPER-LEATHERMAN • Chemistry and Biochemistry Department,
Fairfi eld University , F airfi eld , C T, U SA
DAVID P. HICKEY • Department of Chemistry , U niversity of Utah , S alt Lake City, U T , U SA
KENISE JEFFERSON • Department of Chemistry and Biochemistry, Center for Nanoscience,
University of Missouri—St. Louis , S t. Louis, M O, U SA
GLENN R. JOHNSON • Hexpoint Technologies , M exico Beach, F L, U SA ; C o-located at the Air
Force Civil Engineer , T yndall AFB, F L, U SA
JELENA R. JOVANOVIĆ • Department of Biochemical Engineering and Biotechnology,
Faculty of Technology and Metallurgy , U niversity of Belgrade , B elgrade, S erbia
JOEL L. KAAR • Department of Chemical and Biological Engineering, U niversity of
Colorado , B oulder , C O , U SA
ZORICA D. KNEŽEVIĆ-JUGOVIĆ • Department of Biochemical Engineering and Biotechnology,
Faculty of Technology and Metallurgy , U niversity of Belgrade , B elgrade, S erbia
HEATHER R. LUCKARIFT • Universal Technology Corporation , D ayton, O H , U SA ;
Co-located at the Air Force Civil Engineer , T yndall AFB, F L, U SA
DUŠAN Ž. MIJIN • Department for Biochemical Engineering and Biotechnology,
Faculty of Technology and Metallurgy , U niversity of Belgrade , B elgrade, S erbia
NENAD B. MILOSAVIĆ • Division of Experimental Therapeutics, Department of Medicine,
Columbia University , N ew York, N Y , U SA
ROSS D. MILTON • Department of Chemistry , U niversity of Utah , S alt Lake City, U T , U SA
SHELLEY D. MINTEER • Departments of Chemistry and Materials Science and Engineering,
University of Utah , S alt Lake City, U T , U SA
MICHAEL J. MOEHLENBROCK • Department of Chemistry , S aint Louis University ,
St. Louis, M O, U SA
JOYEETA MUKHERJEE • Chemistry Department, I ndian Institute of Technology Delhi ,
New Delhi, I ndia
JASMINE M. OLVANY • Chemistry Department, L ebanon Valley College , A nnville, P A, U SA
RADIVOJE M. PRODANOVIĆ • Department of Biochemistry, Faculty of Chemistry,
University of Belgrade , B elgrade, S erbia
ix
x Contributors
MICHELLE RASMUSSEN • Chemistry Department, L ebanon Valley College , A nnville,
PA, U SA
DEBRA R. ROLISON • U.S. Naval Research Laboratory , S urface Chemistry Branch ,
Washington, D C, U SA
IPSITA ROY • Department of Biotechnology , N ational Institute of Pharmaceutical Education
and Research (NIPER) , P unjab, I ndia
JULIA L. RUTHERFORD • Chemistry Department, L ebanon Valley College , A nnville,
PA, U SA
OLGA V. SHULGA • ICL Performance Products , S t. Louis, M O, U SA
ANDREA B. STEFANOVIĆ • Department of Biochemical Engineering and Biotechnology,
Faculty of Technology and Metallurgy , U niversity of Belgrade , B elgrade, S erbia
KEITH J. STINE • Department of Chemistry and Biochemistry, Center for Nanoscience,
University of Missouri—Saint Louis , S aint Louis, M O, U SA
ALICE H. SUROVIEC • Department of Chemistry and Biochemistry , B erry College , M t. Berry,
GA , U SA
GAIGE R. VANDEZANDE • Chemistry Department, L ebanon Valley College , A nnville,
GA , U SA
DUŠAN VELIČKOVIĆ • Department of Biochemistry, Faculty of Chemistry,
University of Belgrade , B elgrade, S erbia
JEAN MARIE WALLACE • Nova Research, Inc. , A lexandria, V A, U SA
MAKOTO YOSHIMOTO • Department of Applied Molecular Bioscience, Yamaguchi University ,
Ube, Japan
Chapter 1
Introduction to the Field of Enzyme Immobilization
and Stabilization
Michael J. Moehlenbrock and Shelley D. Minteer
Abstract
Enzyme stabilization is important for many biomedical or industrial application of enzymes (i.e., cell-free
biotransformations and biosensors). In many applications, the goal is to provide extended active lifetime at
normal environmental conditions with traditional substrates at low concentrations in buffered solutions.
However, as enzymes are used for more and more applications, there is a desire to use them in extreme
environmental conditions (i.e., high temperatures), in high substrate concentration or high ionic strength,
and in nontraditional solvent systems. This chapter introduces the topic enzyme stabilization and the
methods used for enzyme stabilization including enzyme immobilization.
Key words Enzyme immobilization , E nzyme stabilization , C ross-linking , E ntrapment , E ncapsulation
1 Introduction
This book details methods for enzyme stabilization and enzyme
immobilization. Although the concepts of enzyme immobilization
and enzyme stabilization are different, they are related. Enzyme
immobilization techniques are strategies focused on retaining an
enzyme on a surface or support. In theory, they are focused on
being able to reuse the enzyme and/or constrain the enzyme to a
particular area of a reactor or device, whereas enzyme stabilization
is focused on extending the active catalytic lifetime of the protein.
Enzyme immobilization is one method for stabilizing enzymes.
Enzymes are proteins with complex and fragile three-dimensional
structures. The main methods for stabilizing enzymes are: protein
engineering , chemical modifi cation, immobilization, and adding
additives for stabilization.
Protein engineering is a very common method for enzyme sta-
bilization, but it is out of the scope of this book, because it is not a
simple or translational method that can be detailed in a chapter.
The common types of protein engineering are directed evolution,
site-directed mutagenesis, and peptide chain extensions. Directed
Shelley D. Minteer (ed.), Enzyme Stabilization and Immobilization: Methods and Protocols, Methods in Molecular Biology,
vol. 1504, DOI 10.1007/978-1-4939-6499-4_1, © Springer Science+Business Media New York 2017
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