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Chitosan in THE PRESERVATION OF AGRICULTURAL COMMODITIES Edited by SILVIA BAUTISTA-BAÑOS Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec-Jojutla, San Isidro Yautepec, Morelos, México GIANFRANCO ROMANAZZI Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, Via Brecce Bianche, Ancona, Italy ANTONIO JIMÉNEZ-APARICIO Departamento de Biotecnología, Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec–Jojutla, San Isidro Yautepec, Morelos, México Amsterdam • Boston • Heidelberg • London New York • Oxford • Paris • San Diego San Francisco • Singapore • Sydney • Tokyo Academic Press is an imprint of Elsevier Academic Press is an imprint of Elsevier 50 Hampshire Street, 5th Floor, Cambridge, MA 02139, USA The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, UK Copyright © 2016 Elsevier Inc. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further infor- mation about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.com/permissions. This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein). Notices Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices, or medical treatment may become necessary. Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein. In using such informa- tion or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility. To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any liability for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein. British Library Cataloguing-in-Publication Data A catalogue record for this book is available from the British Library Library of Congress Cataloging-in-Publication Data A catalog record for this book is available from the Library of Congress ISBN: 978-0-12-802735-6 For information on all Academic Press publications visit our website at http://store.elsevier.com/ Typeset by Thomson Digital Printed and bound in USA LIST OF CONTRIBUTORS Selene Aguilera-Aguirre División de Estudios de Posgrado e Investigación-Alimentos, Instituto Tecnológico de Tepic, LIIA, Laboratorio de Biotecnología, Lagos del Country, Tepic, Nayarit, México Waldo M. Argüelles Monal Grupo de Investigación en Biopolímeros, CTAOA, Centro de Investigación en Alimentación y Desarrollo AC, Hermosillo, Sonora, México Mohamed E.I. Badawy Department of Pesticide Chemistry and Technology, Faculty of Agriculture, Alexandria University, Alexandria, Egypt Laura Leticia Barrera Necha Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec-Jojutla, San Isidro Yautepec, Morelos, México Silvia Bautista-Baños Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec-Jojutla, San Isidro Yautepec, Morelos, México Malick Bill Postharvest Technology Group, Department of Crop Sciences, Tshwane University of Technology, Pretoria West, Pretoria, South Africa Elsa Bosquez-Molina Departamento de Biotecnología, Universidad Autónoma Metropolitana-Iztapalapa Vicentina, Iztapalapa, Mexico City, México Alejandra Chacón-López División de Estudios de Posgrado e Investigación-Alimentos, Instituto Tecnológico de Tepic, LIIA, Laboratorio de Biotecnología, Lagos del Country, Tepic, Nayarit, México Zormy N. Correa-Pacheco Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carreterra Yautepec–Jojutla, San Isidro Yautepec, Morelos, México Mario Onofre Cortez-Rocha Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Hermosillo, Sonora, México Erica Feliziani Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, Via Brecce Bianche, Ancona, Italy Francisco M. Goycoolea Valencia Grupo de Investigación en Biopolímeros, CTAOA, Centro de Investigación en Alimentación y Desarrollo AC, Hermosillo, Sonora, México xi xii List of Contributors Porfirio Gutiérrez-Martínez División de Estudios de Posgrado e Investigación-Alimentos, Instituto Tecnológico de Tepic, LIIA, Laboratorio de Biotecnología, Lagos del Country, Tepic, Nayarit, México Mónica Hernández-López Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec-Jojutla, San Isidro Yautepec, Morelos, México Cynthia Nazareth Hernández-Téllez Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Hermosillo, Sonora, México Antonio Jiménez-Aparicio Departamento de Biotecnología, Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec–Jojutla, San Isidro Yautepec, Morelos, México Lise Korsten Plant Sciences and Crop Sciences, University of Pretoria, Hillcrest, Pretoria, South Africa Jaime Lizardi-Mendoza Grupo de Investigación en Biopolímeros, CTAOA, Centro de Investigación en Alimentación y Desarrollo AC, Hermosillo, Sonora, México Susana Patricia Miranda-Castro Departamento de Ciencias Biológicas, Facultad de Estudios Superiores Cuautitlan, Laboratorio de Biotecnología, UNAM, Estado de México, México Maura Rojas Pirela Departamento de Biología, Facultad de Ciencias, Laboratorio de Enzimología de Parásitos, Universidad de Los Andes, Mérida, Venezuela Maribel Plascencia-Jatomea Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Hermosillo, Sonora, México Entsar I. Rabea Department of Plant Protection, Faculty of Agriculture, Damanhour University, Damanhour, Egypt Miguel Ángel Ramírez-Arrebato UCTB Los Palacios, Instituto Nacional de Ciencias Agrícolas (INCA), Los Palacios, Pinar del Río, Cuba Anelsy Ramos-Guerrero División de Estudios de Posgrado e Investigación-Alimentos, Instituto Tecnológico de Tepic, LIIA, Lagos del Country, Tepic, Nayarit, México Francisco Rodríguez-González Departamento de Biotecnología, Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec-Jojutla, San Isidro Yautepec, Morelos, México List of Contributors xiii Aida Tania Rodríguez-Pedroso UCTB Los Palacios, Instituto Nacional de Ciencias Agrícolas (INCA), Carretera La Francia, Los Palacios, Pinar del Río, Cuba Gianfranco Romanazzi Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, Via Brecce Bianche, Ancona, Italy Dharini Sivakumar Department of Crop Sciences, Postharvest Technology Group, Tshwane University of Technology, Pretoria West, Pretoria, South Africa María Elena Sotelo-Boyás Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec-Jojutla, San Isidro Yautepec, Morelos, México Keith Thompson Department of Crop Sciences, Postharvest Technology Group, Tshwane University of Technology, Pretoria West, South Africa; Cranfield University, Cranfield, Bedford, UK Cristóbal Lárez Velásquez Grupo de Polímeros, Departamento de Química, Facultad de Ciencias, Universidad de Los Andes, Mérida, Venezuela Rita Velázquez-Estrada División de Estudios de Posgrado e Investigación-Alimentos, Instituto Tecnológico de Tepic, LIIA, Laboratorio de Biotecnología, Lagos del Country, Tepic, Nayarit, México Elsa Ventura-Zapata Departamento de Biotecnología, Centro de Desarrollo de Productos Bióticos (CEPROBI), Instituto Politécnico Nacional, Carretera Yautepec-Jojutla, San Isidro Yautepec, Morelos, México Luis Angel Xoca-Orozco División de Estudios de Posgrado e Investigación-Alimentos, Instituto Tecnológico de Tepic, LIIA, Laboratorio de Biotecnología, Lagos del Country, Tepic, Nayarit, México Leonor Zavaleta-Mejía Departamento de Ingeniería Química y Bioquímica, Instituto Tecnológico de Zacatepec, Zacatepec de Hidalgo, Zacatepec, Morelos, México PREFACE In June 2014, the First International Meeting, with the title Chitosan and Its Applications on the Preservation of Agricultural Commodities was held in Morelos, México. This outstanding event brought together relevant scientists and students working with chitosan in the fields of agriculture from different countries including Cuba, Italy, South Africa, Venezuela, and México. This meeting made clear the necessity of summarizing and updating available information on this compound associated within agriculture areas, specifi- cally in dealing with its application for the conservation of horticultural commodities. Regarding this compound it is important to note that over the past de- cade, chitosan (the deacetylated form of chitin) polysaccharides have gained enormous importance in diverse scientific disciplines (e.g., pharmaceutical and biomedical areas), where literature has been systematically published. Furthermore, in these two disciplines, applications of chitosan now include among others, nutrition supplements, drug delivery, gene therapy, tissue en- gineering, and wound healing. Fortunately, advancements have shown posi- tive results also in the area of agriculture. The presence of amino groups (–NH ) in its chemical structure has resulted in chitosan exhibiting unique 2 and ideal properties in different agricultural systems, including food con- servation and food security through development of biodegradable edible coatings and films containing natural antimicrobials. Chitosan has also been integrated into programs of biological disease control due to its document- ed antimicrobial properties for controlling pathogenic microorganisms in various horticultural commodities. It has elicitor properties that enhance the natural defenses of fruits, vegetables, and grains. Chitosan is also be- ing considered in microdevices to be integrated in “intelligent” and active packing for extending fruit and vegetable shelf life. Chitosan can be extracted from diverse marine organisms, insects, and fungi. It has been considered a biodegradable and biocompatible material, not associated with toxicity or side effects. Presently, the use of chitosan has been technologically justified in sustainable agriculture programs because it raises no public health and safety concerns. In the fresh produce indus- try, this compound is safe for the consumer and the environment, and it has been approved by the US Food and Drug Administration (FDA) as a “generally recognized as safe” (GRAS) food additive. Likewise, regulation xv xvi Preface EU 2014/563 included chitosan chloride as the first member of a basic substance list of plant protection products (planned with Regulation EU 2009/1107). This specialized book on chitosan includes the following sections: The first section, Chitosan Obtention and New Materials-Based Chitosan, explores the close relationship between the chemical characteristics of chi- tosan with its main functional properties, and the current functional chito- san derivatives. It explains the relationship between chitosan structure and its modifications and the specific properties in the final product as well as its potential of application, particularly in the agricultural and horticultural food sectors. The second section, Biological Activity, and Mode of Action of Chi- tosan, examines the effects of novel integrated applications of chitosan coat- ings, alone and in combination with other technologies such as modified atmosphere packaging, plant derivatives, plant gums, physical treatments, organic polymers, organic salts, and antagonist microorganisms in the pres- ervation of fresh fruits, vegetables, and grains, including minimally processed products. The enzymatic and microscopic defense mechanisms against plant diseases and pests following chitosan application are also considered. The third section, Biochemical and Molecular Aspects of Chitosan, deals with the main mechanisms proposed to explain the beneficial effects observed when chitinous materials that are employed to control fungal diseases in plants. These effects include the occurrence of a fertilizer effect, indirect inhibition of the pathogens via their decomposition by-products, stimulating/sup- porting growth of profitable microorganisms, and elicitor activity of chitin. Likewise, studies of the gene expression during the chitosan–Colletotrichum– avocado interaction are presented. The fourth section, Chitosan Bionanocom- posites, gives insight on the use of chitosan nanocomposites in biological models associated with fruit and vegetable conservation. Aspects related to the development of chitosan bionanocompounds, main nanoparticle ob- taining methods, and environmental implications associated with the use of nanomaterials in the agricultural area were also considered. The contributors to this book are nationally and internationally rec- ognized scientists with wide experience in the study of different aspects of chitosan. Throughout recent years they have been active researchers in the field of chitosan, focusing in different aspects of agriculture and food con- servation, which is demonstrated by their numerous high-impact publica- tions on chitosan properties and applications. We editors acknowledge and thank their enthusiastic interest in participating in the writing of this book. Preface xvii We are also deeply grateful for the continuous support and encourage- ment of this idea from the Elsevier team: Patricia Osborne (Acquisitions Editor), Jackie Truesdell (Editorial Project Manager), and Susan Li (Produc- tion Department). The Editors Silvia Bautista-Baños Gianfranco Romanazzi Antonio Jiménez-Aparicio CHAPTER 1 Chemical Characteristics and Functional Properties of Chitosan Jaime Lizardi-Mendoza, Waldo M. Argüelles Monal, Francisco M. Goycoolea Valencia INTRODUCTION Chitin is an aminated polysaccharide biosynthesized in several invertebrate animal species. It is a main compound of the exoskeleton of arthropods, the most abundant animal phyla that include insects and crustaceans. The pres- ence of chitin in some microorganisms such as bacteria, fungi, protozoan, and algae species has also been reported [1]. Chitin is considered one of the most abundant biopolymers and is regarded as one of the substances with highest production and degradation rate in the biosphere. Therefore, chitin plays an important role in the biogeochemical cycles of carbon and nitro- gen, mainly in aquatic ecosystems [1,2]. Despite chitin being almost as abundant as cellulose and sharing several chemical and functional features with it, chitin applications are restricted to a few specialized fields. There are several reasons for this; one is that the natural structures of chitin that could be easily used are scarce (in contrast with cellulose structures such as wood or cotton). Other reasons are the difficulties in obtaining stable chitin solutions or the product variability associated with the inherent diversity of the sources and extraction proce- dures. Therefore, most of the chitin production is dedicated to obtain chi- tosan, which is a deacetylated derivative. After the deacetylation p rocess, the resulting chitosan has diverse functional groups: some capable of being ionized, the amino moieties, and also the remaining acetamide groups that are prone to form hydrophobic associations. This chemical characteristic of chitosan has influence in many functional properties of this molecule (i.e., at acid pH values, the amino groups become cationic, promoting the dissolution of chitosan). Also the polycationic character of chitosan al- lows it to interact with diverse types of molecules. This, together with its structural capacities, biocompatibility, and other properties, make chitosan attractive for producing functional materials applicable in several fields. Chitosan in the Preservation of Agricultural Commodities Copyright © 2016 Elsevier Inc. http://dx.doi.org/10.1016/B978-0-12-802735-6.00001-X All rights reserved. 3 4 Chitosan in the Preservation of Agricultural Commodities CHITOSAN: DEFINITION AND SOURCES Chitosan is the term used to denominate the polymer of glucosamine and N-acetyl glucosamine where the deacetylated units are present in major proportion or their distribution in the polymer chain is such that allows it to be dissolved in aqueous diluted acid solutions. A distinctive feature of the chemical structure of chitosan (Figure 1.1) is the predominant presence of units with amino groups that can be ionized. These groups become cationic in acidic media promoting the chitosan dissolution and polyelectrolyte be- havior in solution. In 1859, Rouget obtained an acid soluble fraction of chitin after boiling it in a concentrated potassium hydroxide solution [3]. This is considered the first scientific report of chitosan. Subsequently, the synthesis and occurrence of chitosan in diverse organisms, mainly fungi, was proved. However, its nat- ural abundance is minimal compared with chitin abundance. Hence, most of the chitosan is produced by thermochemical deacetylation of chitin. Multiple procedures have been proposed and developed to obtain chitosan and comprehensive reviews on the theme are available [3–5]. Most of the existing methods to produce chitosan are variations on thermo-alkaline deacetylation of chitin using hydroxides at high temperatures (e.g., >80°C). The industrial production of chitosan today is based on this type of process. Chitosan could be also obtained by homogeneous chemical deacetylation of chitin [6]. Alternatively, biotechnological procedures have been proposed, including extraction from cultures of selected fungi strains or enzymatic deacetylation of chitin with limited success [7,8]. Typical chitin deacetylation by a thermo-alkaline procedure is a het- erogeneous phase reaction, where the process conditions (e.g., starting ma- terial quality, particle size, reactants mixture ratio, additives, agitation rate, etc.) have a determinant influence on the characteristics of the produced chitosan. Consequently, notable variability on the functional properties of produced chitosan has been observed [9]. To achieve uniform production with proper quality control, an accurate monitoring through the entire process is required. Nowadays, consistent quality values and standardized Figure 1.1 Chemical Structure of Chitosan. Schematic interpretation of the polycationic state of chitosan (acetylation = 1/7) in acid aqueous solutions.

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