Chitinandchitosan

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Current Status and New Perspectives on Chitin and Chitosan as Functional

Biopolymers
Article in Applied Biochemistry and Biotechnology · April 2017

DOI: 10.1007/s12010-016-2286-2
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Appl Biochem Biotechnol
DOI 10.1007/s12010-016-2286-2
Current Status and New Perspectives on Chitin

and Chitosan as Functional Biopolymers
Tuyishime Philibert 1 & Byong H. Lee 1,2,3 &

Nsanzabera Fabien 1
Received: 13 July 2016 / Accepted: 10 October 2016

# Springer Science+Business Media New York 2016
Abstract The natural biopolymer chitin and its deacetylated product chitosan are found
abundantly in nature as structural building blocks and are used in all sectors of human
activities like materials science, nutrition, health care, and energy. Far from being fully
recognized, these polymers are able to open opportunities for completely novel applications
due to their exceptional properties which an economic value is intrinsically entrapped. On a
commercial scale, chitosan is mainly obtained from crustacean shells rather than from the
fungal and insect sources. Significant efforts have been devoted to commercialize chitosan
extracted from fungal and insect sources to completely replace crustacean-derived chitosan.
However, the traditional chitin extraction processes are laden with many disadvantages. The
present review discusses the potential bioextraction of chitosan from fungal, insect, and
crustacean as well as its superior physico-chemical properties. The different aspects of fungal,
insects, and crustacean chitosan extraction methods and various parameters having an effect on
the yield of chitin and chitosan are discussed in detail. In addition, this review also deals with
essential attributes of chitosan for high value-added applications in different fields and
highlighted new perspectives on the production of chitin and deacetylated chitosan from
different sources with the concomitant reduction of the environmental impact.
Keywords Biological approach . Chitin . Chitosan . Fungus . Insects . Shellfish wastes
* Byong H. Lee
byong.lee@mail.mcgill.ca
Tuyishime Philibert
tuyiphilbert@gmail.com
1
School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China
2
Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 24341,
South Korea
3
Department of Microbiology/Immunology, McGill University, Montreal, QC H9X3V9, Canada
Introduction
Chitin and its derivative chitosan have been receiving increasing attention as invaluable
biopolymers; they can be found as supporting materials in many aquatic organisms,
terrestrial organisms, and some microorganisms and also present in vertebrates (zebrafish
gut and epithelial cells of fish scales), but the function is not known [10, 87, 90, 123].
Competing with cellulose and starch for being the most abundant natural organic com-
pound, chitin and chitosan are considered to be materials of great futuristic potential with
immense possibilities for structural modifications to impart desired properties and func-
tions and to increase their innumerable applications in diverse scientific areas. In addition,
the water-insoluble property of chitin as highly basic polysaccharide, is probably due to
intra- and intermolecular hydrogen bonds with adjacent –NH or –OH functions groups
formed by the oxygen of acetamido group and the degree of N-acetylation has a striking
effect on insolubility and limits swelling properties of chitin in water compared with
cellulose [45, 48, 119]. For these reasons, researchers have transformed these complex
substances into low MW oligosaccharides known as chitooligosaccharides with low vis-
cosity and relatively small molecular sizes which in turn make them water soluble and
readily absorbed in vivo [45, 81, 114]. Moreover, physicochemical characteristics of chitin
and chitosan vary from batch to batch due to seasonality of raw materials, the quality of
shell, species present, climate, and difficulties in process control; this has led to expand
interest in biotechnology research regarding the production of chitin and chitosan from
different sources such as exoskeleton of insects and cell wall of fungi, etc. [26, 60, 69].
Traditional methods for commercial preparation of chitosan from their resources present
some drawbacks since it is expensive, has inconsistent molecular weight (MW) and degree of
acetylation (DA), and environmentally unfriendly [35, 40, 56]; however, biotechnology
fermentation processes, i.e., deproteination and demineralization by protease and organic acid
bacteria and deacetylation by chitin deacetylase were a suitable and economical method to
extract chitin and their derivatives with recovery of bioactive compounds; these processes can
offer new perspectives for the production of highly viscous chitosan, with a promising
potential for innumerable application [21, 56].
The invaluable chitosan and oligosaccharide have increased due to their versatile biological
activities which can be employed in food, agriculture, medicine, pharmaceutical, personal care
product, and environmental sectors [40, 47]. To further extend their application, the signifi-
cance of their diversity and the applicability of the different forms of these remarkable
biopolymers have yet to be determined; thus, a wealth of knowledge continuously being
added to the exoskeleton of many arthropods including insects and crustaceans and the cell
wall of fungi which are the key sources for these natural biopolymers and also marine
biotechnology as a new field that involves discovery and invention of processing methods
for these materials into valuable commercial products is needed [56, 90, 119].
Chitosan from crustacean as a food industry waste is economically feasible, especially if it
includes the recovery of compounds such as carotenoid pigment (astaxanthin), protein, fish
meal additive in aquaculture, and nutraceuticals which are beneficial in human health promo-
tion [12, 106, 107]. Therefore, chitosan unquestionably can be promoted and marketed as a
green product. Data published on worldwide chitosan market was estimated to be 13,730
metric tons in 2010, but the demand for chitosan is growing rapidly where the market is
expected to reach 40,465 metric tons annually by 2018 [73]. In this review, we have reviewed
the most significant and updated information on biotechnological processes for chitin recovery
from terrestrial, microbial, and marine organism as well as other information on closely related
by-products, which will endeavor to provide a better understanding of various advances in the
ecologically safe extraction of this important biopolymer.
Structural Properties of Chitin and Chitosan
Chitin is called poly-β-[1, 4]-N-acetyl-d-glucosamine or poly(N-acetyl D-glucosamine)

and occurs in three polymorphic forms α-, β-, and γ-chitins, due to very strong
hydrogen bonding between the amide groups and the carbonyl groups of the nearby
chains. Its deacetylated chitosan is a linear polysaccharide composed of randomly
distributed β-(1–4)-linked D-glucosamine and N-acetyl-d-glucosamine; with varying
degree of deacetylation (DD; 40–99 %) and MW [81, 113], the chitosan differ not only
in molecular mass and degree of deacetylation but also by their crystalline, moisture, and
protein content [5, 96]. Their degree of deacetylation is one of the most important factors
influencing both chemical (solubility, flexibility, polymer conformation, viscosity, crys-
tallinity, high surface area, porosity, tensile strength, conductivity, photoluminescence)
and biological properties (biodegradability, biocompatibility, mucoadhesion, hemostatic,
analgesic, adsorption enhancer, antimicrobial, anticholesterolemic, and antioxidant)
which vary with process conditions [14, 96, 122]. In addition, the chitosan-free proton-
ated amino groups makes it possible to form complexes with negatively charged deriv-
atives, such as anionic synthetic polymers, polysaccharides, proteins, dyes, and lipids,
and also with cholesterol, fat, enzymes, tumor cells, bacteria cell wall proteins, or DNA
and RNA. It has also the ability to chelate with various metal ions, due to the neutral or
negatively charged hydroxyl groups of D-glucosamine, GlcN residues, and amino groups
[18, 24, 117]. One of the advantages of chitosan is its insolubility at neutral and acidic
conditions due to its positive charges (NH3+) at pH (pH <6.5). Additionally, alkali
concentration, chemical form of the polymer, type of chitin and polymorph type, chitin
particle size, heterogeneous or homogeneous N-deacetylation, single or multiple process,
atmosphere, and incubation time have been reported to influence chitin and its
deacetylated chitosan properties [5, 53, 98]. The combination of all these and other
characteristics with its good mechanical properties, its biocompatibility, and its biode-
gradability open the way to many novel applications [81].
Processes of Chitin and Chitosan Production
Chemical Process
Ideally, the invaluable production of chitin and their derivatives from resources should be
simple, economical, time saving, and ecofriendly which presides various feature of final
chitosan such as purity, DA, MW, and polydispersity index that largely impact its
applications in various domains. Chemical methods are currently being applied in com-
mercial scale [26, 56]. These industrial techniques normally consist of two steps,
deproteinization by alkali treatment and demineralization by acidic treatment under high
temperature, followed by decolorization step which focuses on removing pigments and
lipids as presented in Fig. 1. In addition, compared with the traditional chitin extraction
Resources of chitin
Chemical process
Shellfish waste Fungi Insect cuticules

Grinding and drying Grinding and drying Grinding and drying Biotechnology process
DEPROTEINIZATION DEPROTEINIZATION DEPROTEINIZATION
Temperature: 65°C - 100°C; temperature: 100°C; 0.75-2.5 N NaOH for 2-
time: 1h – 72 h; concentration: 1.0 M
concentration: 0.125–2.5M of 42 h at 400C -1000C
NaOH; time: 3 h;
NaOH, Na2CO3, KOH, Proteins
K2CO3, Ca (OH) 2, Na2SO3. Proteins Shellfish waste
Proteins DEMINERALIZATION
DECOLOURATION 1-2 N HCl for 0.3-96h at
Organic solvent: ethyl 25 to 1000C
DEMINERALIZATION alcohol. Bleaching
25–2M HCl, HCl, HNO3, with KMnO4, NaOCl
H2SO4, CH3COOH, and Minerals Washing, Drying
H2O2; temperature: and Crushing
HCOOH at 0–1000C for 1–48 h. ambient; time: 3x1h CaCO3, CaPO4
DECOLOURATION
Minerals (CaCO3, CaPO4) Ethyl alcohol
temperature: ambient;
Demineralization
time: 0.5 h - 4 h using lactic acid
DECOLOURATION bacteria
Organic solvent: ethyl
alcohol, acetone or Pigments Minerals
diethyl ether and
CaCO3, CaPO4
Pigments Grinding Deproteinization Deproteinization
Bleaching with Grinding and
with commercial using
KMnO4, NaOCl H2O2; add water
temperature: 20°C – CHITIN enzymes proteolytic bacteria
60°C; time: 0, 25 h – 12
h Pigments, proteins
Enzyme
deactivation
Heterogeneous Homogeneous
Depolymerization/ CHITIN
deacetylation deacetylation
40%–50% NaOH partial hydrolysis 40%–50% NaOH
Deacetylation using
chitindeactylase or lactic
Dissolve in acid
CHITOSAN acid bacteria
(2% acetic acid)
Washing and drying
Hydrolysis using
Chain Substitution CHITOSAN chitinolytic enzymes
Lyophilize Filter elongation
Washing and drying Hydrolysis using

Depolymerization/ chitinolytic enzymes
Chitosonium Precipitate partial hydrolysis
acid salts with NaOH Washing and drying
(water soluble)
Chitosan
Chitosan oligomers
(Free amine form)
Fig. 1 Flow chart of chemical and biological processes for chitin, chitosan, and COS
method, the repeated string with 3 % of NaClO for 10 min before demineralization and
deproteination was suggested as new method appears to be propitious process regarding its
time and energy saving for quick chitin extraction from marine resources [40, 61, 122].
Due to covalent associations with other shell constituents, harsh acid treatments, high
NaOH concentrations, and high deproteinization temperatures may cause hydrolysis of the
polymer, partial DA, undesirable deacetylation, and depolymerization of chitin thus
leading to inconsistent physiological properties in the end products which are a source
of pollution. Additionally, the demineralization of chitin results in detrimental effects on
the molecular mass and the DA that negatively affects the intrinsic properties of the
purified chitin [26, 40]. To avoid the risk of chitin depolymerization, ambient temperature
and stirred bioreactors have been reported to improve the quality and allow the process to
be shorter [9, 120].
Biological Process
Recently, bioextraction of chitin and chitosan has been gaining interest because of different
challenges of chemical processes such as energy consuming, extremely hazardous, unsuit-
able protein components as animal feed, and threat to the environment due to the high
concentrations of mineral acid and caustic employed [56, 121]. This has led to growing
interest to optimize isolation process by minimizing chitin degradation and invoke impu-
rities to the level of invaluable application. Recently, biotechnological processes (Fig. 1)
have also been focusing on the production of high-value commercial by-products such as
chitooligosaccharides (COS), chitinases, lactic acid, antioxidants, valuable protein hydro-
lyzates, carotenoids (astaxanthin), and solubilized calcium ions. Furthermore, some of
these bioactive compounds have been identified to possess nutraceutical potentials that
are profitable to the human health. Therefore, the development of new technologies in
exploration of novel bioactive compounds from marine processing wastes will alleviate
costs associated with its safe disposal. [40, 87, 122].
Extraction of Chitin and Chitosan
Extraction from Insect
Insects are a promising source of new biomass because of their constitution (rich in
proteins, fats, and biopolymers), their cultivation possibilities (grow on organic-
biological waste streams), and with high percentage of dry matter (20–60 %) [29]. The
procedure for extraction of chitin and chitosan from the cuticle of insects is similar to that of
crustacean sources; however, their demineralization is stronger than that of aquatic crusta-
cean material process (Fig. 1). The crystallinity of chitin increases and 55 % of the N-acetyl
groups of silkworm chitin were removed after treatment with 2 N HCl at 100 °C, therefore,
the treatment of insect cuticle with dilute HCl has a binary functionality. The deproteination
of insect pupa and larva using NaOH under mild condition improve the yield of chitin up to
15 % with similar chemical structure and physiochemical properties of commercial α-chitin
from shrimp and chitin structure of the exoskeleton of seven species such as Ailopus
simulatrix, Ailopus strepens, Duroniella fracta, Duroniella laticornis, Oedipoda miniata,
Oedipoda caerulescens, and Pyrgomorpha cognata varied between 5.3 and 8.9 %, and
therefore, this lead them as an alternative chitin source [65, 77]. Dineshet et al. (2014)
reported that 24 mg/1 g of chitosan was successfully extracted using 1 % HCl and 4 % of
NaOH to extract chitosan from American cockroach. Using Fourier transform infrared
spectroscopy (FTIR), this chitosan showed a characteristic bond for β-1-4-glycosidic
linkage [27]. Chitin and chitosan extracted from six different aquatic invertebrate species,
its dry weights, and chitins productivity varied between 5 and 20 % and 66 and 74 %,
respectively. Moreover, Hydrophilus piceus has shown the highest dry weight and chitosan
productivity and Ranatra linearis and Notonecta glauca have shown higher chitin and
chitosan thermal stabilities, respectively. Additionally, by scanning weight (SEM), ther-
mogravimetric analysis (TGA), X-ray diffraction (XRD), and FTIR, α-chitin was observed
with surface morphology of nanofibre structures and the crystalline index value between
76.4 and 90.6 %. These six aquatic invertebrate species may be used as alternative chitin
and chitosan sources for various technological purposes [62].
The four grasshopper species exhibited a quantity of chitin with dry weight ranging from
4.71 to 11.84 %; the male chitin surface structure contained 25–90-nm-wide nanofibers and
90–250-nm nanopores, and in contrast, the elemental analysis, thermal properties, and crys-
talline index values for chitin were similar in males and females. Compared with commercial
chitin digestion rate, both sexes did not show a big difference rate (94.95 % and 88.45–95.48),
respectively [66]. The dry bat guano from bat species Rhinolophus hipposideros had 28 % α-
chitin with a chitosan yield of 79 % and the crystallinity of chitin 85.49 % and 58.51 % of
chitosan was calculated. It has been reported that bat guano can be considered an alternative
source of chitin and chitosan to crab, shrimp, crayfish, and krill [67]. Additionally, chitin
nanofibers of some insects are very similar with those of other crustacean, but the widths of
chitin nanofibers extracted from Melolontha were observed to be much higher than those of
nanofibers isolated from crab, prawn, and fungi species [63]. Schistocerca gregaria had the
highest yield of chitin at 12.2 % compared with Calosoma rugosa and Apis mellifera, and
these insects show a good characteristic of DD, water-binding capacity (WBC) and fat-binding
capacity (FBC), ash content, and moisture content compared with shrimp. This suggests that
exoskeleton of insect should be sited as an alternative chitin source [80]. Moreover, the dry
weight chitin content of adult potato beetle and larva were 20 and 7 %, with a chitosan yield of
72 and 67 %, respectively, therefore the adult potato beetle can be utilized as an alternative
chitin and chitosan source [60]. Recently, the production of chitin and chitosan from insect
sources has drawn increased attention, six different common insect species such as Shistocerca
gregarea, Forsskal, green bugs, German cockroach, Vespid wasp, and Fuessly have been
documented to be sources of chitin with an effective comparison with crustacean, and its
chitins exhibited similar chemical structures and physiological properties. [8].
Exoskeleton of seven species such as D. laticornis, D. fracta, Aiolopus simulatrix, Aiolopus
strepens, O. miniata, O. caerulescens, and P. cognata were investigated; the dry weight chitin
contents of the grasshopper species varied between 5.3 and 8.9 %, and the chitins from seven
orthoptera species had low molecular weights between 5.2 and 6.8 kDa. With that, the
grasshopper could be an alternative source of chitin [65]. Insects and other arthropods make
up the largest and most biodiverse group of organisms on the planet. Likewise, the magnitude
of the chemical diversity which they produce and utilize is also one of the most impressive in
the living world. Insects are still a very promising endeavor, and it is evident that insects are the
richest and most unexplored reservoirs of potentially useful substances as resources for natural
product drug discovery and bioprospecting in general.
Microbial Extraction
To date, thousands of tons of microorganism waste biomass of yeast, bacteria, fungi, and algae
are rich in various kinds of bioactive compounds, such as biopolymers, proteins, lipids, and
pigments, among others. The production of these bioactive compounds led to bring various
biotechnological/fermentation processes into economical valuability [26, 56]. The production
of biopolymers from microbial sources appears to be promising because the process can be
handled to obtain a pure uniform product with a specific characteristic. In addition, the
fermentative production of fungi on low industrial by-products and wastes is just an unlimited,
and in principle, a very economical source of chitin and chitosan. The feasibility of obtaining
β-glucan from the myceliar chitosan/glucan complex and contemporary isolation of chitin and
chitosan make the microbial as invaluable sources of biopolymers using biotechnological
process [54, 88].
Extraction from Fungi and Mushrooms
Fungal cell walls are composed of polysaccharides and glycoproteins. The extractions of chitin
and chitosan from different species of mushrooms such as Pleurotus sajor-caju, Lentinula
edodes, Agaricus bisporus, Auricula-judae, Trametes versicolor, Armillaria mellea, Pleurotus
ostreatus, and Pleurotus eryngii have been studied; the total yields of chitin and chitosan from
these species were around 85–196 and 10–40 mg/g of dried biomass, respectively, under
submerged fermentation, and the cultivation conditions, while type and mainly species of the
mushroom, influence the growth rate of mushrooms. The processes and conditions for
extraction of chitin and chitosan from mushrooms were nearly the same. However, using acid
extraction process, the extraction from mushrooms is difficult and the yields were found to be
low due to complexes of chitin/chitosan with glucans or other polysaccharides [69, 72].
Fungal chitosans have a degree of deacetylation of 70–90 % with an average MW of 1–
2 × 105 Da depending on mushroom species and treatment. The yield of chitosan 120 mg/l and
6.18 g/kg under liquid and solid-state fermentation conditions, respectively, is produced from
L. edodes. Therefore, chitin/chitosan yield depends on mushroom species, harvesting time,
extraction processes, and conditions [26, 56]. This was conformed previously that the highest
amount of extractable chitosan (93.4 mg/200 ml substrate) was obtained using Gongronella
butleri USDB 0201 strain of Zygomycetes; however, the highest yield of chitosan per unit
myceria mass was obtained from Cunninghamella echinulata at 7.14 % [69].
Ospina Álvarez et al. [92] reported that chitin from Ganoderma lucidum fungus had
lower thermal stability than chitin obtained from other common sources such as shells of
crustaceans; however, it showed the highest crystallinity. The biomass from mushrooms
obtained by biotechnological culture is a promising source for obtaining chitin and its
by-products [92].
Fungal chitosan has advantages due to the uniformity of particle size and antimicrobial
effect that has served with water cleaning procedures, beer-brewing process, wound coverage,
and textile production. In addition, under mild condition, media optimization and green
conversion of agro industrial wastes, chitin and chitosan are obtained from mushrooms and
fungi (Table 1) [54, 87]. In this context, production and purification of chitin and chitosan from
the cell walls of waste fungal mycelium offer the advantage of being environmentally friendly
and provide greater potential for a consistent product and also the treatment with HCl or
decolorizing agent and deacetylation step are not required for the extraction of chitosan from
mushrooms and fungi. Additionally, β-glucan can also be isolated from the mycelia chitosan/
glucan complex which has important applications in biomedicine [87].
In the USA alone, mushroom production results in nearly 50,000 metric tons of mushroom
waste material per year with no suitable commercial application; these huge amount of wastes
can be potentially used for the extraction of the high value-added product, chitosan; the
European Food Safety Authority (EFSA) approved that chitosan extracted from Aspergillus
niger is food additive [26, 87].
Cultivation Conditions
Microbial chitin and chitosan productions could be affected by different factors such as
medium composition like yeast extract, peptone, and glucose (YPG) medium, molasses salts
medium (MSM; yeast extract and molasses as carbon source), potato dextrose broth, potato
extract and dextrose (PDB) and YM media (malt extract, polypeptone, and glucose),
Table 1 Amount of chitin, chitosan produced by the different fungi
Microorganism Biomass Chitin Chitosan %DD References
Cunninghamella elegans 16.00 g/L 72.29 mg/g 33.13 mg/g 80 [17]

Rhizopus arrhizus 24.60 g/L 83.20 mg/g 49.31 mg/g 82 [17]
Mucor circinelloides 20.7 g/L 500 mg/g 64 mg/g 83 [32]
C. elegans UCP/WFCC 0542 16.95 g/L ND 2.14 g/L 75.25 [28]
Aspergillus terreus CBNRKR KF529976 24.83 g/L 344.8 mg/g 48.32 mg/g 98 [105]
A. terreus 2.8 mg/L 58 mg/g 83.06 [75]
Ganoderma lucidum 21.87 ± 2.2 g L − 1. 413 mg/g ND ND [92]
C. elegans RCMB 012002, Mucor rouxii RCMB 015002, Rhizopus sp. ND ND 640, 440, 240 mg/l 80.30, 81.50, 80.30 [84]
G. lucidum 19.72 ± 1.06 g/L ND 83.23 ± 4.53 mg/g 80.29 [82]
Mucor indicus 0.09 g/g sugar ND 0.45 g/g cell wall ND [108]
Absidia butleri NCIM 977 8.4 g/L ND 683 mg/L 79.89 [116]
Fomitopsis pinicola ND 30.11 % 71.75 % 73.1 %. [57]
M. rouxii, Aspergillus niger, Aspergillus fumigatus, and Penicillium sp ND 530, 330, 240, 430 mg/g 280, 75, 50, 170 mg/g ND [74]
ND not determined
temperature, pH, aeration, agitation, size and age of producing microbial cells, and growth
time. Depending on fermentation techniques such as solid-state fermenter (SSF) and sub-
merged fermenter (SmF) and microorganism under examination, each factor might have
various effects on chitin/chitosan production. Published data on the percent yields of chitin,
chitosan, glucan, and dry mycelia weight was reported to be 10.1–44 %, 1.2–25.0 %, 2.05–
93.4 %, and 16.8–26.7 %, respectively. Additionally, the kinetics of chitosan production seems
to have a slightly better profile in a continuous reactor. Consequently, chitosan extraction
procedure and fermentation time are important factors affecting chitosan yield and its molec-
ular weight [40, 69, 122].
Fermentation Techniques
Generally, SmF technology has been used to produce chitinous compounds; however, in some
cases, SSF is considered a promising route for chitin and chitosan production and also batch
fermentation was usually used for chitin/chitosan production. As compared with continuous
and batch cultures, fermentation by continuous culture in the stirred tank reactor produced the
highest amount of chitosan from Absidia coerulea 14076, resulting in approximate 3-fold
increase in chitosan production [56, 76].
SSF is an alternative technology for chitin/chitosan production, but there are
inconsistent results on its efficacy in the production of chitosan. The growth of the
fungus G. butleri USDB 0201 using various nitrogen sources under SmF condition
resulted in 1.5–2.5 times higher yields of chitosan compared with SSF; however, the
yield of chitosan by SSF of Lentinus edodes was better by about 6.2 g/kg, 50 times
higher than that obtained from other fungi. Therefore, the efficiency of fermentation
type used depends on microbial source and must be probed for each fungus of
interest. In both cases, SSF seems to be the preferred method in production of low
acetylation degree chitosan where SSF in several parameters have been shown to be
important factors affecting the fungal growth and chitosan production [26, 91].
Erlenmeyer flasks, rotary drum bioreactors, and roux bottles as laboratory model
bioreactors have been selected to evaluate the influence of bioreactors design and
hydrodynamic conditions by SSF technology on chitin and chitosan production. SSF
has shown its potential for chitosan production, but it remains to be exploited
commercially.
Occurrence of Microbial Chitin and Chitosan
Chitin is less distributed in the microbial world than aquatic organism; properly, the main
chitin is eminent in the fungal cell wall and septa of Ascomycete, Zygomycete, Basidio-
mycete, and Deuteroycatas thus serving as a strengthening fibrous element responsible for
cell wall rigidity. It has been reported that 32.7 and 9.4 % of the cell wall of filamentous
forms of the fungus exists as chitin and chitosan, respectively. Moreover, lower amounts of
chitin and chitosan were available in the cell wall of yeast cells (27.9 and 8.4 %,
respectively) compared with those of the filamentous form [26, 54, 87]. A family of
integral membrane proteins called chitin synthases is responsible for the synthesis of
linear chains of chitin β-1, 4-N-acetylglucosamine from the substrate uridine diphos-
phate-N-acetyl glucosamine. In fungus cell wall, polysaccharide biosynthesis of chitin
does not occur by itself, and it is often accompanied by other polysaccharides notably
glucan which provides its special properties useful for many applications (Fig. 2). In
addition, chitosan, cellulose, and other β-glucans (80–90 %) and proteins (3–20 %) are
considerable ingredients of the cell wall, whereas lipids, pigments, and inorganic salts are
present in minor proportions. Zygomycete, particularly Mucor rouxii, Mucor mucedo, and
Mucor circinelloides seem to have higher chitin content in the cell wall. Other promising
resources of chitin are Ascomycetes and Basidiomycetes which contain significant quan-
tities of chitin and acidic polysaccharides as cell wall component (26–65 %) [90, 37] and
Mucoralean strain like Syncephatastrum racemosum or Cunninghamella echinulate,
whose cell wall containing a rather higher amount of chitosan has attracted more attention
to further research on more productive sources of chitin [11, 69].
Physicochemical Properties and Commercial Importance of Microbial

Chitin and Chitosan
Microbial chitin is usually associated with beta-glucans and melanins providing its beneficial
properties and unprecedented properties. In comparison with conventional natural fibers,
fungal chitin-based filamentous structure can be applied directly for many biomedical and
pharmaceutical applications as wound dressing and carriers of micro-encapsulated drugs, so
only relatively low chemical treatments are required during processing [50, 115]. The quality
of chitosan varies with DD of the N-acetyl groups, MW, purity, manufacturing process, color,
clarity, consistency, and uniformity. However, microbial chitosans generally show a lower DD
than those of commercial crab shell chitosans, making them highly allure for valuable
applications. Based on structure-property relationship, microbial chitosan is effective and
feasible in many applications (Table 3) [24, 42]. In addition, chitosan with low MW appears
to reduce the tensile strength and elongation of the chitosan membrane, increasing its
permeability thus it could be used as thread or membrane in many medical applications and
as a powder in cholesterol absorption.
UDP-Galp
UDP- UDP-Glc UDP- UDP-

GLcNAc Galp Man
Chitin synthase β-(1,3)-glucan synthase Mannosyl transferase

Galactosamine
Galactopyran
Chitin β-(1,3)- β-(1,5)- α-(1,2)/α-(1,6)

glucan galactofuran mannan
β-(1,3;1,4)-glucan
Galactomannan Galactominogalactan
β-(1,6)-branched β-(1,3)-glucan
Fig. 2 Flow chart of the synthesis and modification steps involved in some fungi cell wall polysaccharide
biosynthesis. Source: Gastebois [37]
Biotechnology Approach
The cultivation of selected fungi has attracted attention as a potential method for the chitin/
chitosan production because the fermentation process can continue throughout the year and
can be manipulated to obtain a product with specific characteristics (Fig. 3) [54].
Based on the variability of the polymer physiochemical properties, more attention has been
focused on improvement of microbial chitin/chitosan production by fermentation process,
genetic modification of proteinase bacteria, and extraction method. These processes might
develop novel biopolymers such as chitin nanofiber of fungal mycelia by chemical, physio-
logical, or biological routes coupled with renewable and biodegradable properties to direct the
growth of selected fungi. This will hopefully lead to the production of truly green nanocom-
posite materials with interesting characteristic in terms of ductility, durability, stability,
strength, etc. [2, 48, 87].
Currently, such integrated processes are a recurring matter in industrial biotechnology
development. Marine invertebrate organisms including sponges such as Aiolochroia crassa,
Aplysina aerophoba, Aplysina cauliformis, Aplysina cavernicola, Aplysina fulva, Spongilla
lacustris, and Aplysina fistularis and in 4 of 15 species of Hexacorallia (Anthozoa), but not in
Octocorallia (Anthozoa) did not only provide alternative sources of α-chitin but some of them
also inspire investigations to develop biomimetic composites, scaffolds, and templates for
practical use in materials science, biomedicine, and tissue engineering [19, 31, 123]. Addi-
tionally, enzyme-aided production is a vital step towards chitosan production in green chem-
istry area as chemical process is engraved with a number of limitations and bottlenecks. As an
alternative, partial enzymatic hydrolysis of chitosan using chitinases or chitosanases has been
studied, but these enzymes lead to the production of heterogeneous chitosan oligomers; other
enzymes like chitin deacetylases (EC 3.5.1.41) from fungus, insects, and marine bacteria and
chitin oligosaccharide deacetylases, chitin oligosaccharide (COD; EC 3.5.1–) can convert into
defined chitin oligomers [95, 128]. However, CDA-producing capabilities of most fungal
strains are low and their fermentation requirements are complicated; therefore, searching for a
more suitable strain of CDA-producing bacteria to replace the current fungal strains is needed.
In addition, physical modification of chitin will increase the efficiency of enzymes involved in
deacetylation for the production of chitosans and the combination enzymes with a specific
pattern NodB deacetylase exclusively the GLcNAc unit at the non-reducing end and COD that
catalyzes deacetylation of N-acetyl-D-glucosamine residues under mild reaction conditions and
results into production of novel superior-quality chitosan could circumvent the problems of
chemical process [2, 43, 55].
Supplement
minerals
Biotechnology products
Fermentation e.g. antibiotic, organic acid,
Carbon resources:
(SmF/SSF) of fungi
e.g. Lingocellusic enzymes, etc.
waste
Fungal autolysate
Autolysis
Solid residue
Chitin deacetylase Cell wall

Chitosan Chitin purification
Fig. 3 Schematic pathway of fungi chitosan by fermentation process

Extraction from Marine Organisms
Marine environment consists of several simple polymers such as chitin, chitosan, colla-
gen, carrageenan, fucoidan, and alginate which play a pre-eminent role in biological and
biomedical applications. Blue biotechnology concerned with the application of molecular
biological methods to marine and freshwater organism is a new emerging area of research
with significant applications [40, 56]. The isolation, characterization, and application of
marine biomaterials has gained a strong market position for novelty, but extensive
research is required to evolve new and improved products at competitive prices [76,
119]. Marine organism crustacean shells are the most important chitin source, its chitin
recovery involves three steps: namely demineralization (DM), deproteination (DP), and
elimination of lipid and pigments (Fig. 1). Traditional methods for chitin production from
crustacean have several problems. As regulations have become stricter now, there is a
need to treat and utilize the waste in a more efficient manner. Therefore, there is a
significant interest regarding recycling of crustacean biowaste. Alternatively, to overcome
the problems of chemical process, different biological processes have been investigated:
bio-demineralization, bio-deproteinization by biological treatments, deproteinization by
enzymes, and the combination of bio-demineralization and bio-deproteinization [35,
122]. In addition, biological production of chitin can reduce environmental pollution
and depolymerization of chitin. The main drawback of the biological process is its lower
efficiency and quality, being time consuming, and thus has a higher cost reported in the
chemical process. Yet, biological production seems ever promising because of possible
recovery of by-products that result in less environmental burden [40, 122]. To reduce
strong solution reagents at high temperature in traditional methods that results in de-
creased quality and also increased costs and ecological problem, much attention has been
given to microwave irradiation; the highest percent of chitosan production at 19.47 %,
89.34 % DD, and 806,931 DA MW were achieved from Persian Gulf shrimp waste by
response surface methodology [89].
Bio-demineralization and Bio-deproteinization by Biological Treatments
Microbial proteases such as Lactobacillus sp., Bacillus sp., Pseudomonas sp., Serrati
marcescens, etc. are the most often applied strains of chitin and chitosan production. Depend-
ing on materials (carbon source), microorganism, fermentation type (auto-fermentation, fer-
mentation in a single-step, successive fermentations, and co-fermentation), and time, as shown
in Table 2, DP values varied up to 97 %, whereas DM values varied up to 99 %. Moreover,
other microbial sources like fungi also have been tried for fermentation; A. niger facilitated the
DP of shrimp-shell powder and the release of hydrolyzed proteins [6, 40]. In addition, organic
acid and proteases produced by microorganisms can dissolve calcium and hydrolyze protein
found in crustacean shell. The efficiency of fermentation depends on factors such as inoculum
levels, shell content in medium, shell size, carbon sources (glucose, sucrose, malt, cassava,
molasses, and date juice), initial pH, and pH during fermentation. The fermentation types
include liquid- and solid-phase cultures in aerobic and anaerobic conditions and the order in
case of successive fermentation consisting of DM and DP processes thus an increase in the
inoculum concentration resulted in improved DM. At industrial scale, rapid acidification of the
medium is very important, especially for preserving shell materials fresh and minimizing a
Table 2 Fermentation-mediated extraction of chitin from marine wastes
Microorganism Waste source Process efficiency (%) References
DP DM DA
Bacillus mojavensis A21 and Bacillus subtilis A26/microbial proteases Shrimp: Metapenaeus monoceros 88 ± 5 ND 90 [124]
Lactobacillus sp. B2 Crab (Callinectes bellicosus) wastes 56 88 95 [34]
Bacillus cereus SV1 Shrimp: M. monoceros 95, 65 67, 15 ND [38]
Bacillus licheniformis RP1 Shrimp waste 81 68 ND [41]
Bacillus pumilus A1/response surface methodology Shrimp; M. monoceros 94 88 ND [39]
Lactobacillus helveticus Shrimp: Parapenaeus longirostris 78 98 ND [6]
Lactobacillus pentosus L7 and proteolytic Bacillus thuringiensis SA Shrimp (Litopenaeus vannamei) 96.8 ± 0.7 ND 98.2 ± 0.5 [99]
Serratia marcescens B742 Shrimp: Penaeus vannamei 91.4 ± 4.6 ND ND [49]
Paenibacillus woosongensis TKB2/bacterial protease cocktail Black tiger shrimp shell wastes 80 ND 82.25 ± 0.07 [97]
Bacillus mojavensis A21, B. subtilis A26, Vibrio metschnikovii J1, Shrimp (M. monoceros) 77 ± 3, 75 ± 3, 65 ± 3, 59 ± 3 ND ND [125]
B. licheniformis MP1
B. cereus and Pseudomonas spp./2-step fermentation Brown crab (Cancer pagurus) 68.9 98.9 82.0 ± 1.69 [44]
Penaeus aeruginosa, S. marcescens and B. pumilus/protease Shrimp shells (Penaeus merguiensis) 74.76 78.46 86 DD [33]
co-fermentation
S. marcescens B742 and Lactobacillus plantarum ATCC 8014/2-step Shrimp; P. vannamei 94.5 93.0 80.17 [127]
fermentation
L. plantarum PTCC 1058 Shrimp shell ND 75 ND [68]
Lactobacillus lactis, Teredinobacter turnirae/co-fermentation Prawn shell 66.5 78.8 ND [7]
Bacillus licheniformis and Gluconobacter oxydans/co-fermentation Shrimp (L. vannamei) 87 93.5 ND [76]
ND not determined
possible reduction of chitin polymers. Medium conditioning and ensilation is an efficient

method for preservation and allowing the recovery of value-added by-products such as
proteins, pigments, and enzymes from crab or shrimp-shell wastes [100, 122].
Liquefaction of the shell waste occurs mainly by proteolytic enzymes produced by starter
microorganisms, Gut bacteria present in the intestinal systems of biomaterials, or proteases
present in the biowaste. The separated liquor represents 60–70 % of the silage and is composed
of proteins and lipids and more than 85 % of the chitin present in the substrate remain in the
sediment. Lactic acid-producing bacilli were mostly adopted for the acidification and decal-
cification processes [56, 105]. Proteases of Bacillus mojavensis A21 was compared with other
microbial proteases of Bacillus subtilis A26, Bacillus licheniformis NH1, B. licheniformis
MP1, Vibrio metschnikovii J1, and Aspergillus clavatus under control of several operating
parameters, such as enzyme/substrate ratio, temperature, and incubation time. These sequential
treatments of chitin extraction allowed the recovery of 18.5 ± 2.3 % of its initial dry mass as
water-insoluble white fibrous material [125]. In addition, enzymatic methods have focused on
processes that lead to the recovery of the three main components, chitin, carotenoid pigments,
and protein thus these treatments were to eliminate the protein present in the waste with the
concomitant production of protein hydrolyzates (as food enhancer ingredient) and raw chitin.
In this process, the commercial enzymes used were: Alcalase 2.4 L, Pancreatin, Delvolase,
Cytolase PCL5, Econase CEPi and MP 100, Maxazime NNP, and Cellupulin MG as well as
non-commercial protease complexes and crude proteases. The effect of proteolytic enzymes
(tuna proteinase, papain, and a bacterial proteinase) on DP of crustacean wastes, the residual
protein associated with the chitin after enzyme treatment, was about 5 %, and the residual
protein levels in the shrimp waste after the DP were 1.3 and 2.8 % for chymotrypsin and
papain-treated sample, respectively, and high enzyme to waste ratio (E/W) was needed for
maximum DP; moreover, the typical value of E/W ratio was 0.7 and 1.0 % (W/W) for
chymotrypsin and papain, respectively [40, 122]. The protein removal from shrimp shell
was not complete with any of the enzyme preparations; thus, they were not capable of
releasing minerals from the protein-chitin complex. In addition, enzymatic DP followed by
microwave-assisted DM allowed the recovery of high quality chitin under environmental
friendly conditions [40, 70]. Furthermore, the highest DP increased from 85 to 90 % when
crab shell waste was treated with 1 % Delvolase. This enzyme showed the highest DP activity
compared with other commercial enzymes such as Cytolase PCL5, Econase CEPi, Econase
MP 1000, Maxazme NNP, and Cellupulin but with no complete removal of the residual protein
associated with the chitin. Stenotrophomonas maltophilia showed the highest DP activity
(82 %) than purified microbial protease (64 %) under the same condition. Lactic acid
fermentation method combined with multistep freeze pump thaw (FPT) proved to be an
effective biological method to avoid excessive depolymerization and loss of crystallinity
during chitosan production, which offers new perspective applications for chitosan [93].
Additionally, promising production of chitin from praw shell using co-fermentation with
lactic acid-producing bacteria and protease-producing bacteria in a batch culture under anaer-
obic condition has been studied. In co-fermentation with at least two different strains for DM
and DP (Table 2) together in a one-batch culture, this kind of problem would be always
encountered. Consequently, the discovery and characterization of novel microorganisms that
proliferate and secreted organic acid and proteases will be necessary in the near future [56, 94].
Successive two-step fermentation has also been challenged to recover a final chitin from
biomaterial, one can use one microorganism that produces both organic acids and proteases or
two different ones (acidification and DM process) for a stable waste ensilation. This was also
confirmed by Pareek et al. [94] using Penicillium oxalicum SAEM-5 to study a bioconversion
of chitin to chitosan in a two-stage chemical and enzymatic process which enhanced degree of
deacetylation with formation of 90 % deacetylated chitosan by 3.2-fold. Moreover, combina-
tion with a protease and acid-producing bacterium B. licheniformis 21886 and Gluconobacter
oxydans DSM-2003 was effective for DP and DM (Table 2) compared with those cultivated
individually. A chitin content of 90.8 % and a total amount of 16 g/L of broth organic acids
were found in the co-fermentation with waste. The major acids were lactic, formic, and acetic
acid, followed by gluconic, succinic, pyroglutamine, and propionic acid [76]. Further studies
on mode of action and catalytic mechanism of the enzyme system along with the exploration
of more effective pretreatment strategies are vital for effective bioconversion of chitosan with a
desired degree of deacetylation.
Application
Chitin and its derivatives as a potential resource as well as multiple functional substrates
ranged from biomedical, pharmaceutical, food, and environmental industries due to its phys-
icochemical and biological properties (Table 3). There are now currently over 2000 concrete
applications of these versatile biopolymers which are non-exhaustively listed in Fig. 4 [24, 71,
104]. The field of nutrition is the most important user of chitosan; in addition, 2288 metric tons
is estimated to be used in 2018 in food and beverages in the USA. However, their applicability
as bioactive compounds and their nutraceutical properties are not clearly described [87].
Identification of nutraceutical potential of natural compounds is a growing field, and marine
processing by-products represent potential feed stocks for this purpose and further future
research developments to implement them for the human health promotion are needed.
Food Application
The use of chitosan in the food industry is well known due to its functional properties.
As there is no complete study on the metabolism of chitin and chitosan in the human
body, the use of these polymers in food processing industries still needs to be further
explored. To increase the application of chitin and chitosan, conversion of processing
discards into valuable by-products and alternative specialty materials is needed for food
research [101, 126]. Chitosan cationic polysaccharide and bioactive nature offers a
great potential as a dietary fiber. These characteristics contribute to faster intestinal
transit time and reduce putrefactive activity that is linked to higher energy recoveries
by the host due to the increased bacterial metabolites in the colon. Chitosan has ability
to lower cholesterol by blocking the absorption of dietary fat and cholesterols [30].
Chitosan products have been shown to facilitate weight and body fat loss in the human
body thus decreased systolic and diastolic blood pressure [83]. Additionally, chitosan
significantly increased the excretion of highly atherogenic saturated fatty acids (lauric
and myristic) compared with other fibers. It may be used to balance fatty acid ratio in
our diet for reducing the risk of many chronic diseases. Chitosan and its two deriva-
tives not only have low cytotoxicity but can control over nutrition and achieve insulin
resistance therapy [78, 110].
Furthermore, chitosan, being a source of dietary fiber, is also a valuable prebiotic
which can promote optimal colonic conditions. The design of functional foods that could
Table 3 Application of chitin and chitosan from different resources
Resources Properties Application References
Shrimp: Parapenaeus longirostris, Penaeus carinatus, and Anticancer activity Pharmaceutical industry, cancer therapy [20, 102, 109]
Penaeus monodon
Shrimp: P. longirostris, Metapenaeus monoceros Antimicrobial activities Food, cosmetics, agriculture, and health [1, 14, 124]
industry
Mushroom: Shiitake stipes Antimicrobial and antitumor activities Pharmaceutical, food, cosmetics, agriculture, [22]
and health industry
P. longirostris Coating properties Food industry, agriculture [13, 15]
Fresh pink shrimp shell waste Nematicidal activity Agriculture [103]
Shrimp: Litopenaeus vanammei and crab: Ucides cordatus Nanomembrane Bionanotechnology [4]
Fungus: Fusarium oxysporum f. sp. lycopersici Nanoparticle Agriculture [111]
Shrimp: M. monoceros Antitumor, antioxidant, and antimicrobial Pharmaceutical industry, agriculture, and [125]
activities cosmetic
Fungus; Cunninghamella elegans, Aspergillus niger Antioxidant activity, antibacteria activities Agriculture [16, 25, 79]
Orthoptera species (Insecta) Antimicrobial and antioxidant activities Food industry [59]
Medicinal fungus (Fomitopsis pinicola) Antimicrobial, antitumor, and antioxidant Pharmaceutical industry [57]
activities
Egyptian shrimp shells, Bryozoa; Plumatella repens Adsorption Water treatment [64, 86]
Fungi: Mucor rouxii, Cuninghamella elegans, Rhizopus sp Antimicrobial activities Pharmaceutical, food, cosmetics, agriculture, [84]
and health industry
The Norway lobster (Nephrops norvegi-cus) Antimicrobial and antiproliferative activities Pharmaceutical industry [112]
The body segments of a diplopod species (Julus terrestris) DNA interaction, antitumor, and antimicrobial Medicine, pharmacy, bioengineering, and [58]
activities water treatment
Insect: larvae and adult Colorado potato beetle Antimicrobial and antioxidant activities Food, cosmetics, agriculture, and health industry [60]
(Leptinotarsa decemlineata)
Shrimp waste Adsorption of metal ions Water pollution treatment [52]
Daphnia longispina ephippia An effective coating properties Food industry, agriculture [60]
Enzyme and whole cell immobilizer

Metabolic analysis of biological fluid (Electronic devices)
Biotechnology
Biosensors
Bioseparation
Antimicrobial (antifungal) agent and biopesticide
Agriculture
Fertilizer and biocontrol agent
Potent antioxidant and matrix metalloproteinase inhibitor
Nerve conduit for nerve regeneration
Potential Substitutes in Acute Burn
Immuno-Stimulating and Anticancer Effects
Biomedicine
Treatment of Tumors and Leukemia, drug and gene delivery,
(Crab, shrimp, squid)
Chitin, chitosan and

Crustacean shells
Aiding Bone Formation, Treating Hypoinsulinaemia

their derivatives
Adjuvant properties and spermicide

Artificial skin, hypertension and ACE-I-inhibitors
Removal and binding of dyes
Removal/recovery of metal ions from wastewaters
Wastewater
Sludge treatment and dehydration agent
treatment Biological denitrification
In lotions
Ingredients for hair and skin care
Cosmetics In creams
Nail lacquers
Photography Film forming ability, nanoimprinting lithography
Opthalmology An ideal contact lens
Food and nutrition

Preservation of food
Filmogenic properties Food wrapping
Food
Filtration and clarification of fruit
Hypolipidemic and hypocholesterolimic
Coating material
Fig. 4 Flow chart of summary of applications of chitin/chitosan and their oligosaccharide
generate specific short chain fat acid (SCFA) patterns at controlled sites in the large
intestine is an interesting challenge aiming for positive health consequences [46].
Chitosan becomes active in the digestive tract by contributing to viscosity, bulk, fat
binding, and fermentative processes. Its use should enhance overall human health and
alleviate current chronic disease problems.
Biomedical Application
Chitin and chitosan show excellent biological properties such as non-toxicity, biodeg-
radation in the human body, biocompatibility and immuno-stimulating, anticancer,
antibacterial effects, wound healing, haemostatic activity in cell culture, tissue engi-
neering, and drug delivery [24]. Chitin is also used as an excipient and drug carrier in
film, gel, or powder form for applications involving mucoadhesivity. In particular,
chitooligosaccharides (COS) and their derivatives are potential candidates capable of
preventing or treating diverse chronic inflammation such as colitis, periodontal dis-
ease, hepatitis, and gastritis and through drug delivery system [3, 36, 51].
Agriculture Application
Synthetic bactericides as the main method for controlling diseases have been a growing
concern over the indiscriminate use on crops because of the possible harmful effects on
human health and the emergence of resistant pathogen [23, 42]. Thus, there is a worldwide
trend to explore new alternatives in order to reduce the use of synthetic chemical agents.
Chitosan and COS have become a promising alternative treatment due to its antimicrobial
activity and in reducing soil-borne diseases. In addition, chitin exhibits several functions,
including retention of nutrients in the soil, and contributes to the nitrogen cycle COS are
known to have eliciting activities leading to a variety of defense responses in plants in
response to microbial infections to reduce the negative impact of diseases on yield and
quality of crops [23, 47].
Bionanotechnology and Other Applications
Chitosan have shown excellent biotechnology application as nanobiodevices. It has been

used in the preparation of graphitic carbon nanocapsules, tungsten carbide, and tungsten
carbides/graphitic carbon composites. Chitin and chitosan have a versatile potential
application in photography, ophthalmology, cosmetics, water and waste treatment, and
in textile industry [88, 118, 120]. The rationale and intelligent use of industrial by-
products from fishery processing towards the production of durable components and easy
to recycle is becoming a must to save the integrity and biodiversity of our planet. The
use of chitin nanofibrils and other natural polymers to produce innovative goods seems
to go in upcoming research [85].
Conclusions and Future Perspectives
Interest in biopolymer chitin and their derivatives from marine processing wastes
(crustacean wastes) by chemical deacetylation continues to grow steadily over the
years. However, chitosan obtained using the current treatments results in inconsis-
tencies which are often accompanied by uncontrollable hydrolysis and chemical
modifications that eventually result in the formation of undesired by-products and
large amounts of aqueous waste. Subsequently, it also escalates the cost and poses
environmental risks. It is against this background that biotechnology process offers an
opportunity to preserve the exceptional qualities of chitin and its derivatives. Further
works based on biotechnology process using a robust bacterium with desired yields of
chitin compared with the chemical method will alleviate costs associated with a safe
and environmentally friendly wastes disposal. Another emerging effective strategy
would be to focus on advances in molecular biology that allow genetic transformation
of fungi to obtain strains with high capability for chitin synthesis and utilizing the
unlimited waste mycelium to overproduce useful recombinant chitin deacetylases in
different cloning hosts for transforming into defined chitin oligosaccharides and also
use insects as the most unexplored reservoirs of potentially useful substances as
resources for natural product drug discovery and bioprospecting in general. Taking
into account its numerous benefits, future research developments to implement bio-
technology process that is still confined to laboratory scale will indeed take a leap
forward thus beingpredicted to be a promising technology that may be eventually seen

in the near future for sustainable chitin production through green chemistry technol-
ogies in commercial-scale development.
Compliance with Ethical Standards
Declaration of Interest The authors report no conflicts of interest. The authors alone are responsible for the
content and writing of this manuscript.
Ethical Statement The article does not contain any studies with human participants performed by any of the
authors.
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Current Status and New Perspectives on Chitin and Chitosan as Functional

Article in Applied Biochemistry and Biotechnology · April 2017

Philibert Tuyishime Byong Lee

11 PUBLICATIONS 541 CITATIONS

6 PUBLICATIONS 220 CITATIONS

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Current Status and New Perspectives on Chitin

Tuyishime Philibert 1 & Byong H. Lee 1,2,3 &

Received: 13 July 2016 / Accepted: 10 October 2016

Keywords Biological approach . Chitin . Chitosan . Fungus . Insects . Shellfish wastes

Structural Properties of Chitin and Chitosan

Chitin is called poly-β-[1, 4]-N-acetyl-d-glucosamine or poly(N-acetyl D-glucosamine)

Processes of Chitin and Chitosan Production

Shellfish waste Fungi Insect cuticules

Washing and drying Hydrolysis using

Extraction of Chitin and Chitosan

Extraction from Insect

Extraction from Fungi and Mushrooms

Microorganism Biomass Chitin Chitosan %DD References

Cunninghamella elegans 16.00 g/L 72.29 mg/g 33.13 mg/g 80 [17]

Occurrence of Microbial Chitin and Chitosan

Physicochemical Properties and Commercial Importance of Microbial

UDP- UDP-Glc UDP- UDP-

Chitin synthase β-(1,3)-glucan synthase Mannosyl transferase

Chitin β-(1,3)- β-(1,5)- α-(1,2)/α-(1,6)

Chitin deacetylase Cell wall

Fig. 3 Schematic pathway of fungi chitosan by fermentation process

Extraction from Marine Organisms

Bio-demineralization and Bio-deproteinization by Biological Treatments

Microorganism Waste source Process efficiency (%) References

possible reduction of chitin polymers. Medium conditioning and ensilation is an efficient

Resources Properties Application References

Enzyme and whole cell immobilizer

Chitin, chitosan and

Aiding Bone Formation, Treating Hypoinsulinaemia

Adjuvant properties and spermicide

Photography Film forming ability, nanoimprinting lithography

Opthalmology An ideal contact lens

Food and nutrition

Fig. 4 Flow chart of summary of applications of chitin/chitosan and their oligosaccharide

Bionanotechnology and Other Applications

Chitosan have shown excellent biotechnology application as nanobiodevices. It has been

Conclusions and Future Perspectives

forward thus beingpredicted to be a promising technology that may be eventually seen

Compliance with Ethical Standards

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