Bacterial Nanocellulose. From Biotechnology to Bio-Economy

244 Pages·2016·18.966 MB·English

by  Miguel Gama, Fernando Dourado, Stanislaw Bielecki

Bacterial NanoCellulose
From Biotechnology to Bio-Economy
Edited by
Miguel Gama
Minho University, Biological Engineering Department
Campus de Gualtar, Braga, Portugal
Fernando Dourado
Minho University, Biological Engineering Department
Campus de Gualtar, Braga, Portugal
Stanislaw Bielecki
Lodz University of Technology, Institute of Technical Biochemistry
Lodz, Poland
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List of Contributors
Hugo Águas i3N/CENIMAT, Department of Materials Science, Faculty of Science and Technology,
Universidade NOVA de Lisboa and CEMOP/UNINOVA, Lisbon, Portugal
Stanislaw Bielecki Lodz University of Technology, Institute of Technical Biochemistry, Lodz,
Poland
Alexander Bismarck Polymer and Composite Engineering (PaCE) Group, Institute of Materials
Chemistry and Research, Faculty of Chemistry, University of Vienna, Vienna, Austria; Polymer and
Composite Engineering (PaCE) Group, Department of Chemical Engineering, Imperial College
London, London, United Kingdom
Rusdianto Budiraharjo Department of Biotechnology, Indonesia International Institute for Life
Sciences (i3L), Jakarta, Indonesia
Son Chu-Ky School of Biotechnology and Food Technology, Hanoi University of Science and
Technology, Hanoi, Vietnam
Ana Cristina Rodrigues Minho University, Biological Engineering Department, Campus de
Gualtar, Braga, Portugal
Fernando Dourado Minho University, Biological Engineering Department, Campus de Gualtar,
Braga, Portugal
Paulo Duarte i3N/CENIMAT, Department of Materials Science, Faculty of Science and Technology,
Universidade NOVA de Lisboa and CEMOP/UNINOVA, Lisbon, Portugal
Gabriella Gita Febriana Department of Biomedicine, Indonesia International Institute for Life
Sciences (i3L), Jakarta, Indonesia
Ana Fontão Minho University, Biological Engineering Department, Campus de Gualtar, Braga,
Portugal
Elvira Fortunato i3N/CENIMAT, Department of Materials Science, Faculty of Science and
Technology, Universidade NOVA de Lisboa and CEMOP/UNINOVA, Lisbon, Portugal
Miguel Gama Minho University, Biological Engineering Department, Campus de Gualtar, Braga,
Portugal
Diana Gaspar i3N/CENIMAT, Department of Materials Science, Faculty of Science and
Technology, Universidade NOVA de Lisboa and CEMOP/UNINOVA, Lisbon, Portugal
xi
List of Contributors
Marzena Jedrzejczak-Krzepkowska Lodz University of Technology, Institute of Technical
Biochemistry, Lodz, Poland
Marek Kolodziejczyk Lodz University of Technology, Institute of Technical Biochemistry, Lodz,
Poland
Tetsuo Kondo Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University,
Japan
Katarzyna Kubiak Lodz University of Technology, Institute of Technical Biochemistry, Lodz,
Poland
Marta Leal Minho University, Biological Engineering Department, Campus de Gualtar, Braga,
Portugal
Koon-Yang Lee The Composites Centre, Department of Aeronautics, Imperial College London,
London, United Kingdom
Falk Liebner Division of Chemistry of Renewable Resources, University of Natural Resources
and Life Sciences Vienna, Tulln, Austria
Karolina Ludwicka Lodz University of Technology, Institute of Technical Biochemistry, Lodz,
Poland
Daniela Martins Minho University, Biological Engineering Department, Campus de Gualtar, Braga,
Portugal
Rodrigo Martins i3N/CENIMAT, Department of Materials Science, Faculty of Science and
Technology, Universidade NOVA de Lisboa and CEMOP/UNINOVA, Lisbon, Portugal
Danh-Nguyen Nguyen School of Economics and Management, Hanoi University of Science and
Technology, Hanoi, Vietnam
Teresa Pankiewicz Lodz University of Technology, Institute of Technical Biochemistry, Lodz, Poland
Luís Pereira i3N/CENIMAT, Department of Materials Science, Faculty of Science and Technology,
Universidade NOVA de Lisboa and CEMOP/UNINOVA, Lisbon, Portugal
Muenduen Phisalaphong Department of Chemical Engineering, Faculty of Engineering, Chulalong-
korn University, Bangkok, Thailand
Ma. Eden S. Piadozo Department of Agricultural and Applied Economics, College of Economics
and Management, University of the Philippines Los Baños, Los Baños, Philippines
Nicole Pircher Division of Chemistry of Renewable Resources, University of Natural Resources
and Life Sciences Vienna, Tulln, Austria
Thomas Rosenau Division of Chemistry of Renewable Resources, University of Natural Resources
and Life Sciences Vienna, Tulln, Austria
xii
List of Contributors
Malgorzata Ryngajllo Lodz University of Technology, Institute of Technical Biochemistry, Lodz,
Poland
Przemysław Rytczak Lodz University of Technology, Institute of Technical Biochemistry, Lodz,
Poland
Siriporn Taokaew Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn
University, Bangkok, Thailand
Tien-Khai Tran School of Economics, University of Economics Ho Chi Minh City, Vietnam
Claudia van den Berg TNO, The Netherlands
António Vicente i3N/CENIMAT, Department of Materials Science, Faculty of Science and
Technology, Universidade NOVA de Lisboa and CEMOP/UNINOVA, Lisbon, Portugal
xiii
Preface
Bioeconomy is based on the efficient use of diverse natural substrates and processes, for
the production of food, feed, bio-based products, and bioenergy. An example of the rational
usage of natural resources is the fabrication of Bacterial NanoCellulose (BNC), which may be
produced from various wastes or biomass.
BNC is a natural polymer, synthesized by a number of species, among which acetic acid
bacteria (AAB) of the species Gluconoacetobacter xylinus and Ga. hansenii are its best
recognized and most efficient producers. These bacteria produce an extracellular, chemically
pure β-glucan, supporting their survival in the natural environment since the cells are kept
at the surface of culture media, being entrapped inside gelatinous, skin-like membranes,
consisting of entangled cellulose fibers. Such self-immobilization of the cells promotes
efficient transport of nutrients and oxygen, which is essential for these aerobic bacteria.
Owing to the high water holding capacity (water accounts for around 98% wet membrane
weight) this polysaccharide protects its producers from desiccation. Cellulose matrix shields
these bacteria also from other adverse environmental factors, like UV radiation. Bacteria
synthesizing cellulose adhere to its surface and are relatively motile, leading to successful
colonization of ecological niches. This in turn reduces available space and supply of nutrients
for other microorganisms populating the same habitats.
BNC synthesized by Gluconoacetobacter species is characterized by unique features,
resulting from its natural biological role, such as high hydrophilicity, crystallinity, purity
and water holding capacity, mechanical durability and resistance to degradation, excellent
biocompatibility and lack of cytotoxicity and allergenicity. Because of these properties and
susceptibility to biological, chemical, and physical modifications, this natural biomaterial
found numerous applications in fabrication of bioproducts and is considered a “bio-base” for
the development of novel materials in various fields, like food processing, electronics, paper
making, chemical and textile industries as well as in medicine. Increasing applications of
bacterial cellulose and its derivatives in various branches of industry and in medicine gave
rise to intensive studies on the improvement of its production efficiency, while attempting to
lower the costs of BC biosynthesis and modifications.
xv
Preface
AAB have a long history of use in several fermentation processes. Their exploitation
gradually emerged in biotechnological applications, especially in the biosynthesis of
useful chemicals and processes for the manufacture of several fermented food products.
Taxonomic studies, from traditional to polyphasic approaches, have gradually allowed the
proper classification of several ABB into distinct genera and species, among them, the BNC
producers, notably G. xylinus. Chapter 1 first reviews the main historical steps involved in the
taxonomic classification of AAB. It then addresses the lying potential behind mixed microbial
fermentations, from kombucha to nata de coco, both sharing in common, the contribution of
cellulose-producing bacteria for the fermentation process.
Recent advances in molecular biology studies on Gluconacetobacter species metabolism
are presented in Chapter 2. Its readers will find the map of metabolic pathways of these
bacterial species, information about utilization of various wastes for BNC biosynthesis,
novel findings related to the structure of cellulose synthase operon and flanking sequences in
Gluconacetobacter species and other microbial cellulose producers as well as the explanation
of roles played by proteins that are encoded by these sequences in cellulose biosynthesis.
Also, crystallographic structures of A and B subunits of cellulose synthase and its complex
with c-di-GMP, which were resolved in 2013 and 2014, are presented in this chapter.
Examples of genetic modifications of Gluconacetobacter species, with particular emphasis on
genetic tools applied, and their effect on BNC biosynthesis are also included.
Chapter 3 describes mechanisms of bacterial cellulose biosynthesis regulation, paving way
to further genetic studies, leading to better comprehension of molecular control of BNC
secretion.
Chapter 4 summarizes analytical techniques that are used to characterize BNC and presents
common physical, chemical methods enabling for a detailed description of the properties of
native and modified BNC.
Chapter 5 describes the intriguing properties of BNC aerogels and the way they can be
obtained. These aerogels are expected to find use in high-performance thermal insulation, as
matrix material for gas separation, carrier for magnetic particles (electro actuators), catalysts,
quantum dots (bio-sensing, volumetric displays), or bioactive compounds (controlled drug
release). BNC aerogels are furthermore promising cell scaffolds (tissue engineering) and
precursor materials for the manufacture of carbon aerogels (electrochemical applications).
BNC is a promising material for the production of high performance renewable composites
because of its high tensile properties, low density, and low toxicity. Chapter 6 starts with
the discussion of both theoretical and experimental tensile properties of nanometre-scale
cellulose fibrils, more commonly known as nanocellulose. This is then followed by what
neat BNC offers as nanoreinforcement for polymers. The tensile properties of various neat
BNC-reinforced polymer nanocomposites published in the literature to date are reviewed and
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Preface
are tabulated. In addition, the micromechanical models that are suitable to describe the tensile
properties of BNC-reinforced polymer nanocomposites are critically discussed.
The use of BNC as a food product, and particularly its potential as a novel food additive,
is reviewed in Chapter 7. Its “technological” potential as a novel hydrocolloid for the
modification of textural properties of food products is addressed. This work briefly reports
on the already commercially available cellulose based hydrocolloids, namely colloidal
microcrystalline cellulose, then reviewing the studies which demonstrate the potential of
BNC in this field. Chapter 8 overviews the European Union (EU) legislative framework of
Novel Foods/Novel Food Ingredients and Food additives, to better familiarize the reader of
the general steps involved in a premarket approval within these regulatory frameworks.
Chapter 9 describes the medical and cosmetic applications of BNC, starting from the most
known never-dried wound dressings and facial masks, going through BNC internal uses
as implantable material, like artificial heart valves, blood vessels or dura mater, and finally
covering the topic of cellulose numerous modifications for the use as, inter alia, substitute
of cartilage, tubes for nerves regeneration, or composites with porous materials for meshes
preparation. Potential usage of this natural biomaterial in other fields of medical sciences, like
tissue engineering exploiting the forms of porous scaffolds, as well as drug delivery sector
applying BNC-based release systems, is also presented.
Chapter 10 provides the readers with the necessary basic knowledge for the implementation
of original, innovative technological solutions, based on results of scientific research. This
knowledge may be used for faster and more precise presentation of data and collection of
suitable documentation for the certification processes that are obligatory for commercial
products, based on BNC. This chapter includes the description of relevant definitions and
classification of medical products, principal requirements, conformity assessment procedures,
obligations of manufacturers and other information related to medical applications of the
biomaterial in order to present readers with a clearer picture of the issues related to obtaining
the necessary certificates for medical devices before placing them onto the market.
Chapter 11 reviews the main applications of BNC in electronics, either as a substrate
(passive) or as a real electronic material (active), and discusses the advantages of BNC in the
field of Paper Electronics.
Chapter 12 explores the process and economics of a computer simulated large scale
production of BNC by static culture conditions. A comparative economic analysis between
modern and traditional plants is not straightforward due to differences in local feedstock
costs, energy, equipments, taxes, labor, currency, and so forth. However, data here gathered
showed that even if considering the use of low cost substrates, the biotechnological
fermentation of BNC is markedly expensive and inefficient, as compared to traditional
fermentation. The high capital investment and high production costs increased by almost two
xvii
Preface
orders of magnitude the selling price of BNC produced in a modern technological set, which
would limit the scope of market penetration.
Finally, Chapters 13 and 14 overview, perhaps for the first time, the nata de coco business in
the Philippines, Thailand, Vietnam, and Indonesia, providing an insight into the current trade
situation, including exports and import analysis, identify the major raw nata producers, their
production practices, marketing outlets, and their selling price. The profitability of growing
raw nata de coco business is also analyzed.
xviii
CHAPTER 1
Taxonomic Review and Microbial Ecology
in Bacterial NanoCellulose Fermentation
Fernando Dourado*, Malgorzata Ryngajllo**, Marzena
Jedrzejczak-Krzepkowska**, Stanislaw Bielecki**, Miguel Gama*
*Minho University, Biological Engineering Department, Campus de Gualtar, Braga, Portugal;
**Lodz University of Technology, Institute of Technical Biochemistry, Lodz, Poland
Acetic Acid Bacteria
Acetic acid bacteria (AAB) are well-known producers of certain foods and drinks, such as
vinegar, kombucha tea, and cocoa. They are also known for being spoilers of other food
products such as wine, beer, soft drinks, and fruits. Cellulose is also a specific product from
AAB metabolism. The AAB name derives from the bacteria’s ability to oxidize ethanol into
acetic acid.
The Acetobacteraceae family consist of a wide group of strictly aerobic, Gram negative,
AAB, endowed with the ability to oxidize a wide variety of carbohydrates, alcohols, and
sugar alcohol into acetic acid and other organic acids (such as gluconic, fumaric, citric,
oxoacids, and ketones) and even amino acids. Among the several genera of this family, the
Acetobacter and Komagataeibacter genus are the most notable acetic acid producers; also
they show rather high tolerance to acidic and alcoholic environments, both scenarios highly
prohibitive of the growth of other microorganisms. Acetobacter strains have a higher capacity
for acetic acid production from ethanol, whereas Komagataeibacter oxidize sugars better
[1–5]. Both genera typically display a diauxic growth curve when cultured in a medium
containing ethanol, the first phase being characterized by ethanol oxidation to acetic acid,
while in a second stage (overoxidation phase) acetic acid is oxidized to water and carbon
dioxide, for further growth [6].
The Taxonomic Classification of Acetic Acid Bacteria
The taxonomic classification of AAB at the species level has been an evolving subject not
only because of the methodologies used but also due to these bacteria’s tendency to undergo
spontaneous mutations. Initially, taxonomic classification was based on morphologic,
Bacterial NanoCellulose
1
http://dx.doi.org/10.1016/B978-0-444-63458-0.00001-9 Copyright © 2016 Elsevier B.V. All rights reserved.