Table Of ContentChitosan 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
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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.
