Enzymes in Fruit and Vegetable Processing: Chemistry and Engineering Applications

Enzymes in Fruit and Vegetable Processing Chemistry and Engineering Applications

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Enzymes in Fruit and Vegetable Processing Chemistry and Engineering Applications

Boca Raton London New York

CRC Press is an imprint of the Taylor & Francis Group, an informa business

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CRC Press Taylor & Francis Group 6000 Broken Sound Parkway NW, Suite 300 Boca Raton, FL 33487-2742 © 2010 by Taylor and Francis Group, LLC CRC Press is an imprint of Taylor & Francis Group, an Informa business No claim to original U.S. Government works Printed in the United States of America on acid-free paper 10 9 8 7 6 5 4 3 2 1 International Standard Book Number-13: 978-1-4200-9434-3 (Ebook-PDF) This book contains information obtained from authentic and highly regarded sources. Reasonable efforts have been made to publish reliable data and information, but the author and publisher cannot assume responsibility for the validity of all materials or the consequences of their use. The authors and publishers have attempted to trace the copyright holders of all material reproduced in this publication and apologize to copyright holders if permission to publish in this form has not been obtained. If any copyright material has not been acknowledged please write and let us know so we may rectify in any future reprint. Except as permitted under U.S. Copyright Law, no part of this book may be reprinted, reproduced, transmitted, or utilized in any form by any electronic, mechanical, or other means, now known or hereafter invented, including photocopying, microfilming, and recording, or in any information storage or retrieval system, without written permission from the publishers. For permission to photocopy or use material electronically from this work, please access www.copyright. com (http://www.copyright.com/) or contact the Copyright Clearance Center, Inc. (CCC), 222 Rosewood Drive, Danvers, MA 01923, 978-750-8400. CCC is a not-for-profit organization that provides licenses and registration for a variety of users. For organizations that have been granted a photocopy license by the CCC, a separate system of payment has been arranged. Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only for identification and explanation without intent to infringe. Visit the Taylor & Francis Web site at http://www.taylorandfrancis.com and the CRC Press Web site at http://www.crcpress.com

Contents Preface................................................................................................................ vii The Editor............................................................................................................ix List of Contributors............................................................................................xi Chapter 1 Introduction to Enzymes............................................................ 1 Alev Bayındırlı Chapter 2 Effect of Enzymatic Reactions on Color of Fruits and Vegetables............................................................................ 19 J. Brian Adams Chapter 3 Major Enzymes of Flavor Volatiles Production and Regulation in Fresh Fruits and Vegetables.......................... 45 Jun Song Chapter 4 Effect of Enzymatic Reactions on Texture of Fruits and Vegetables............................................................................ 71 Luis F. Goulao, Domingos P. F. Almeida, and Cristina M. Oliveira Chapter 5 Selection of the Indicator Enzyme for Blanching of€Vegetables............................................................................. 123 Vural Gökmen Chapter 6 Enzymatic Peeling of Citrus Fruits...................................... 145 Maria Teresa Pretel, Paloma Sánchez-Bel, Isabel Egea, and€Felix Romojaro Chapter 7 Use of Enzymes for Non-Citrus Fruit Juice Production................................................................................. 175 Liliana N. Ceci and Jorge E. Lozano

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Contents

Chapter 8 Enzymes in Citrus Juice Processing.................................... 197 Domenico Cautela, Domenico Castaldo, Luigi Servillo, and€Alfonso Giovane Chapter 9 Use of Enzymes for Wine Production.................................. 215 Encarna Gómez-Plaza, Inmaculada Romero-Cascales, and€Ana Belén Bautista-Ortín Chapter 10 Effect of Novel Food Processing on Fruit and Vegetable Enzymes.......................................................... 245 Indrawati Oey Chapter 11 Biosensors for Fruit and Vegetable Processing................. 313 Danielle Cristhina Melo Ferreira, Lucilene Dornelles Mello, Renata Kelly Mendes, and Lauro Tatsuo Kubota Chapter 12 Enzymes in Fruit and Vegetable Processing: Future Trends in Enzyme Discovery, Design, Production, and Application........................................................................ 341 Marco A. van den Berg, Johannes A. Roubos, and Lucie Parˇenicová Index................................................................................................................. 359

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Preface Fruits and vegetables are consumed as fresh or processed into different type of products. Some of the naturally occurring enzymes in fruits and vegetables have undesirable effects on product quality, and therefore enzyme inactivation is required during fruit and vegetable processing in order to increase the product shelf-life. Commercial enzyme preparations are also used as processing aids in fruit and vegetable processing to improve the process efficiency and product quality, because enzymes show activity on specific substrates under mild processing conditions. Therefore, there has been a striking growth in the enzyme market for the fruit and vegetable industry. While fruit and vegetable processing is the subject of many books and other publications, the purpose of this book is to give detailed information about enzymes in fruit and vegetable processing from chemistry to engineering applications. Chapters are well written by an authoritative author(s) and follow a consistent style. There are 12 chapters in this book, and the chapters provide a comprehensive review of the chapter title important to the field of enzymes and fruit and vegetable processing by focusing on the most promising new international research developments and their current and potential industry applications. Fundamental aspects of enzymes are given in Chapter 1. Color, flavor, and texture are important post-harvest quality parameters of fruits and vegetables. There are a number of product-specific details, dependent on the morphology, composition, and character of the individual produce. Chapters 2, 3, and 4 describe in detail the effect of enzymes on color, flavor, and texture of selected fruits and vegetables. Selection of the indicator enzyme for blanching of vegetables is summarized in Chapter 5. For enzymes as processing aids, Chapter 6 describes in detail the enzymatic peeling of citrus fruits and Chapter 7 presents the importance of enzymes for juice production from pome, stone, and berry fruits. Inactivation of enzymes is required to obtain cloudy juice from citrus fruits. Chapter 8 is related to citrus juices; orange juice receives particular attention. Enzymes also play an important role in winemaking. The application of industrial enzyme preparations in the wine industry is a common practice. The use of enzymes for vii

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wine production is the focus of Chapter 9. Chapter 10 provides serious review of the inactivation effect of novel technologies on fruit and vegetable enzymes to maximize product quality. Chapter 11 presents both chemical and technological information on enzyme-based biosensors for fruit and vegetable processing. The literature reported in each chapter highlights the current status of knowledge in the related area. Future trends for industrial use of enzymes are discussed in Chapter 12. The conclusion part of each chapter also presents the reader with potential research possibilities and applications. This book is a reference book to search or learn more about fruit and vegetable enzymes and enzyme-based processing of fruit and vegetables according to the latest enzyme-assisted technologies and potential applications of new approaches obtained from university and other research centers and laboratories. Such knowledge is important for the companies dealing with fruit and vegetable processing to be competitive and also for the collaboration among industry, university, and research centers. This book is also for the graduate students and young researchers who will play an important role for future perspectives of enzymes in fruit and vegetable processing. Alev Bayındırlı

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The Editor Alev Bayındırlı is a professor in the Department of Food Engineering, Middle East Technical University, Ankara, Turkey. She has authored or co-authored 30 journal articles. She received a BS degree (1982) from the Department of Chemical Engineering, Middle East Technical University. MS (1985) and PhD (1989) degrees are from the Department of Food Engineering, Middle East Technical University. She is working on food chemistry and technology, especially enzymes in fruit and vegetable processing.

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List of Contributors J. Brian Adams Formerly of Campden & Chorleywood Food Research Association (Campden BRI) Chipping Campden Gloucestershire, UK Domingos P.F. Almeida Faculdade de Ciências Universidade do Porto Porto, Portugal Escola Superior de Biotecnologia Universidade Católica Portuguesa Porto, Portugal Ana Belén Bautista-Ortín Departamento de Tecnología de Alimentos, Nutrición y Bromatología Facultad de Veterinaria, Universidad de Murcia Campus de Espinardo Murcia, Spain Alev Bayındırlı Middle East Technical University Department of Food Engineering Ankara, Turkey

Inmaculada Romero-Cascales Departamento de Tecnología de Alimentos, Nutrición y Bromatología Facultad de Veterinaria, Universidad de Murcia Campus de Espinardo Murcia, Spain Domenico Castaldo Stazione Sperimentale per le Industrie delle Essenze e dei Derivati dagli Agrumi (SSEA) Reggio Calabria, Italy Domenico Cautela Stazione Sperimentale per le Industrie delle Essenze e dei Derivati dagli Agrumi (SSEA) Reggio Calabria, Italy Liliana N. Ceci Planta Piloto de Ingeniería Química UNS-CONICET Bahía Blanca, Argentina Isabel Egea Departamento Biología del Estrés y Patología Vegetal Centro de Edafología y Biología Aplicada del Segura-CSIC Espinardo, Murcia, Spain xi

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List of Contributors

Danielle Cristhina Melo Ferreira Institute of Chemistry Unicamp Campinas, São Paulo, Brazil

Lucilene Dornelles Mello UNIPAMPA Campus Bagé Bagé, RS, Brazil

Alfonso Giovane Dipartimento di Biochimica e Biofisica Seconda Università degli Studi di Napoli Napoli, Italy

Renata Kelly Mendes Institute of Chemistry Unicamp Campinas, São Paulo, Brazil

Vural Gökmen Department of Food Engineering Hacettepe University Ankara, Turkey Encarna Gómez-Plaza Departamento de Tecnología de Alimentos, Nutrición y Bromatología Facultad de Veterinaria, Universidad de Murcia Campus de Espinardo Murcia, Spain Luis F. Goulao Secção de Horticultura Instituto Superior de Agronomia Lisbon, Portugal Centro de Ecofisiologia, Bioquimica e Biotecnologia Vegetal Instituto de Investigação Cientifica Tropical Oeiras, Portugal Lauro Tatsuo Kubota Institute of Chemistry Unicamp Campinas, São Paulo, Brazil Jorge E. Lozano Planta Piloto de Ingeniería Química UNS-CONICET Bahía Blanca, Argentina

Indrawati Oey Department of Food Science University of Otago Dunedin, New Zealand Cristina M. Oliveira Secção de Horticultura Instituto Superior de Agronomia Lisbon, Portugal Lucie Parˇ enicová DSM Biotechnology Centre, DSM Delft, The Netherlands Maria Teresa Pretel Escuela Politécnica Superior de Orihuela Universidad Miguel Hernández Alicante, Spain Felix Romojaro Departamento Biología del Estrés y Patología Vegetal Centro de Edafología y Biología Aplicada del Segura-CSIC Espinardo, Murcia, Spain Johannes A. Roubos DSM Biotechnology Centre, DSM Delft, The Netherlands

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List of Contributors Paloma Sánchez-Bel Departamento Biología del Estrés y Patología Vegetal Centro de Edafología y Biología Aplicada del Segura-CSIC Espinardo, Murcia, Spain Luigi Servillo Dipartimento di Biochimica e Biofisica Seconda Università degli Studi di Napoli Napoli, Italy

xiii Jun Song Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Nova Scotia, Canada Marco A. van den Berg DSM Biotechnology Centre, DSM Delft, The Netherlands

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chapter one

Introduction to enzymes Alev Bayındırlı Contents 1.1 Nature of Enzymes.................................................................................... 1 1.2 Enzyme Classification and Nomenclature............................................. 2 1.3 Enzyme Kinetics........................................................................................ 3 1.4 Factors Affecting Enzyme Activity......................................................... 7 1.5 Enzyme Inactivation............................................................................... 10 1.6 Enzymes in Fruit and Vegetable Processing........................................ 13 Abbreviations.................................................................................................... 16 References........................................................................................................... 16

1.1â•… Nature of enzymes Enzymes are effective protein catalysts for biochemical reactions. The structural components of proteins are L-α-amino acids with the exception of glycine, which is not chiral. The four levels of protein structure are primary, secondary, tertiary, and quaternary structures. Primary structure is related to the amino acid sequence. The amino group of one amino acid is joined to the carboxyl group of the next amino acid by covalent bonding, known as a peptide bond. The amino acid side-chain groups vary in terms of their properties such as polarity, charge, and size. The polar amino acid side groups tend to be on the outside of the protein where they interact with water, whereas the hydrophobic groups tend to be in the interior part of the protein. Secondary structure (α-helix, β-pleated sheet, and turns) is important for protein conformation. Right-handed α-helix is a regular arrangement of the polypeptide backbone by hydrogen bonding between the carbonyl oxygen of one residue (i) and the nitrogenous proton of the other residue (i+4). β-pleated sheet is a pleated structure composed of polypeptide chains linked together through interamide hydrogen bonding between adjacent strands of the sheet. Tertiary structure refers to the threedimensional structure of folded protein. Presence of disulfide bridges, hydrogen bonding, ionic bonding, and hydrophobic and van der Waals interactions maintain the protein conformation. Folding the protein brings 1

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together amino acid side groups from different parts of the amino acid sequence of the polypeptide chain to form the enzyme active site that consists of a few amino acid residues and occupies a relatively small portion of the total enzyme volume. The rest of the enzyme is important for the three-dimensional integrity. The quaternary structure of a protein results from the association of two or more polypeptide chains (subunits). Specificity and catalytic power are two characteristics of an enzyme. Most enzymes can be extremely specific for their substrates and catalyze reactions under mild conditions by lowering the free energy requirement of the transition state without altering the equilibrium condition. The enzyme specificity depends on the conformation of the active site. The enzyme-substrate binding is generally explained by lock-and-key model (conformational perfect fit) or induced fit model (enzyme conformation change such as closing around the substrate). The lock-and-key model has been modified due to the flexibility of enzymes in solution. The binding of the substrate to the enzyme results in a distortion of the substrate into the conformation of the transition state, and the enzyme itself also undergoes a change in conformation to fit the substrate. Many enzymes exhibit stereochemical specificity in that they catalyze the reactions of one conformation but not the other. Catalytic power is increased by use of binding energy, induced-fit, proximity effect, and stabilization of charges in hydrophobic environment. The catalytic activity of many enzymes depends on the presence of cofactor for catalytic activity. If the organic compound as cofactor is loosely attached to enzyme, it is called a coenzyme. It is called a prosthetic group when the organic compound attaches firmly to the enzyme by covalent bond. Metal ion activators such as Ca++, Cu++, Co++, Fe++, Fe+++, Mn++, Mg++, Mo+++, and Zn++ can be cofactors. An enzyme without its cofactor is called an apoenzyme. An enzyme with a cofactor is referred as a haloenzyme. Enzymes catalyze the reactions by covalent catalysis or general acid/base catalysis.

1.2â•… Enzyme classification and nomenclature Enzymes are classified into six groups (Table€1.1) according to the reaction catalyzed and denoted by an EC (Enzyme Commission) number. The first, second, and third–fourth digits of these numbers show class of the enzyme, type of the bond involved in the reaction, and specificity of the bond, respectively. Systematic nomenclature is the addition of the suffix -ase to the enzyme-catalyzed reaction with the name of the substrate. For example, naringinase, and α-L-Rhamnoside rhamnohydrolase are trivial and systematic names of the enzyme numbered as EC 3.2.1.40, respectively. Some of the enzyme-related databases are IUBMB, International Union of Biochemistry and Molecular Biology enzyme no menclature (www.chem.qmul. ac.uk/ iubmb/enzyme/); BRENDA, comprehensive enzyme information system

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Chapter one:â•… Introduction to enzymes

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Table€1.1╇ Classification of Enzymes EC Class

Reaction Catalyzed −

−

EC1: Oxidoreductases

A +B A+B

EC2: Transferases

AB + C  A + BC

EC3: Hydrolases

EC4: Lyases

AB + H 2O  AH + BOH

X

Y

A − B  A = B+ X −Y

EC5: Isomerase

X

Y

Y

X

A− B  A− B EC6: Ligases

A + B  AB

Examples Peroxidase Catalase Polyphenol oxidase Lipoxygenase Ascorbic acid oxidase Glucose oxidase Alcohol dehydrogenase Amylosucrase Dextransucrase Transglutaminase Invertase Chlorophyllase Amylase Cellulose Polygalacturonase Lipase Galactosidase Thermolysin Pectin lyase Phenylalanine ammonia lyase Cysteine sulfoxide lyase Hydroperoxide lyase Glucose isomerase Carotenoid isomerase Hydroxycinnamate CoA ligase

(www.brenda-enzymes.org); the ExPASy, Expert Protein Analysis System enzyme nomenclature (www.expasy.org/enzyme/); and EBIPDB, European Bioinformatics Institute–Protein Data Bank enzyme structures database (www.ebi.ac.uk/thornton-srv/databases/enzymes).

1.3â•… Enzyme kinetics Besides the quasi-steady-state kinetics (Briggs and Haldane approach), the rate of enzyme catalyzed reactions is generally modeled by the Michaelis-Menten approach. For a simple enzymatic reaction, binding of substrate (S) with free enzyme (E) is followed by an irreversible

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breakdown of enzyme-substrate complex (ES) to free enzyme and product (P). The substrate binding with E is assumed to be very fast relative to the breakdown of ES complex to E and P. Therefore, the substrate binding is assumed to be at equilibrium as shown in the following reaction scheme: k



1 2   → ES k →E + P E+S ←  k −1



(1.1)

The Michaelis-Menten approach concerns the initial reaction rate where there is very little product formation. It is impossible to know the enzyme concentration in enzyme preparations. Therefore, the amount of the enzyme is given as units of activity per amount of sample. One international enzyme unit is the amount of enzyme that produces 1 micromole of product per minute. According to the applied enzyme activity assay, the enzyme unit definition must be clearly stated in research or application. The total enzyme amount (Eo) equals the sum of the amount of E and ES complex. In terms of amounts, it can be represented as follows:

CEo = CE + CES

(1.2)



The dissociation constant (Km), which is also called the MichaelisMenten constant, is a measure of the affinity of enzyme for substrate:



Km =

k−1 CECS = k1 CES



(1.3)

If the enzyme has high affinity for the substrate, then the reaction will occur faster and Km has a lower value. High Km value means less affinity. Km varies considerably from one enzyme to another and also with different substrates for the same enzyme. For these elementary reactions 1.1, the initial reaction rate or reaction velocity (v) is expressed as

v = k2CES

(1.4)



If the enzyme is stable during the reaction, the maximum initial reaction rate (vmax) corresponds to

vmax = k2CEo



(1.5)

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Chapter one:â•… Introduction to enzymes

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If the initial concentration of substrate (So) is very high during the reaction (CSo >> CEo), the concentration of substrate remains constant during the initial period of reaction (CSo ≈ CS). Combining Equations 1.2–1.5, the Michaelis-Menten equation is obtained as v=

vmaxCS K m + CS

(1.6)

As an example, kinetic properties of polygalacturonase assayed in different commercial enzyme preparations were studied and the reactions in all samples followed Michaelis–Menten kinetics (Ortega et al., 2004) Michaelis-Menten expression can be simplified as follows:



zero order expression : v = vmax first order expression : v =



for C s >> K m

vmax C Km S

(1.7)



for C s