Extraction & Characterization Chitosan from

different fungi strain

Dr Swati Nandi Chakraborty, Anuruna Biswas

ABSTRACT
Chitosan is a complex natural hydrophilic polysaccharide. Chitosan, a flexible natural hydrophilic polysaccharide,
is composed of glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) units linked by beta 1, 4-glycosidic
bonds. Chitin, which is found in the exoskeletons of crustaceans such as prawns, lobsters, crabs, insects, and
fungal cell walls, is the primary ingredient. Conventional method of chitosan production has harmful
environmental effects since it produces millions of tons of basic and acidic residues, which are discharged into the
ecosystem without any Further treatment. The enzymatic deacetylation of chitin is the major mechanism for
synthesis of chitosan in fungi. The chitosan produced from crustacean shell waste had inconsistent
physicochemical properties because of variable and seasonal supply of raw materials and difficulties in processing
methods. To overcome these problems, fungi have been treated as an alternative source for chitosan production.
This review focuses on the isolation of chitosan-producing fungi from soil settings, including strains such as
Mucor rouxii and Rhizopus oryzae, and describes the chitosan extraction processes, which include
deproteinisation, deacetylation and acid precipitation. The physicochemical parameters of fungal chitosan are
determined using analytical methods such as FTIR, SEM, and XRD. Chitosan derived from soil fungi exhibits
superior solubility, deacetylation, and biocompatibility compared to conventional sources, rendering it ideal for
biomedical applications such as drug delivery and wound healing. The paper emphasis the potential of soil fungi
as an environmentally sustainable and efficient source of chitosan, along with opportunities for further research
and industrial scalability.

KEYWORDS: chitosan, fungi, extraction, characterization, biopolymer, deacetylation, cell wall

Introduction
In its natural state, chitosan is a linear polymer of 1,4-glycosidically linked glucosamine (2-
amino-2-deoxy-D-glucopyranose), which is a mucopolysaccharide. In actuality, though, it
also frequently includes acetylglucosamine (2-acetamido-2-deoxy-D-glycopyranose) residues.
If a substance contains less than 50% of these acetylglucosamines, it is classified as chitosan;
if it contains 50% or more, it is classified as chitin. With at least 1010 tons found in the
biosphere, chitin (also known as -1,4-N-acetyl-D-glucosamine) is the most prevalent natural
biopolymer after cellulose. Additionally, chitin is a part of the cell walls of fungi, insects, and
crustaceans’ exoskeletons. It has recently become clear how important chitin is to marine life
as a source of carbon and nitrogen, as well as how it affects the ocean’s ecosystem.
Shrimp, crab, squid, and marine crustacean bone plates are the main sources of chitosan and
chitin used in industrial production. The polymers chitin and chitosan are biocompatible,
biodegradable, and nontoxic. They have a huge amount of untapped potential to help make
sustainable agriculture a reality. One of the most researched natural polymers in a variety of
application areas is chitosan. For instance, its hemostatic, antimicrobial, biodegradable, and
biocompatible qualities make it useful in tissue engineering. It can also be utilized as a starting
material to create hydrogels, membranes, sponges, and electrospun fibers. When chitosan is used
as a base material, it enhances the mechanics and biocompatibility of other materials, including
micro and nanoparticles, active ingredients, and other biopolymers, thereby creating an
environment that is conducive to cell regeneration. Because of its mechanical, permeability,
bactericidal, antimicrobial, and nontoxic qualities, chitosan is also used in food packaging.
Because it is a naturally occurring active absorbent that can be used to absorb heavy metal ions,
chitosan is used in bioremediation. Because chitosan contains active groups, it can be altered by
 adding substances like carbon nanotubes, humic acids, and the algae Ulva lactuca. This
improves chitosan’s ability to absorb metal ions. In actuality, new-generation supercapacitor
electrodes are made from porous carbon microspheres derived from chitosan. Furthermore,
because of its many qualities, chitosan has a lot of potential in the agricultural sector. As a
proteinase inhibitor, chitosan promotes the formation of calloses and aids in the biosynthesis of
phytoalexins. When applied topically, chitosan improves stomatal conductance, raises the
amount of abscisic acid (ABA), and decreases sweating in plants without changing biomass,
height, leaf area, or root height. Additionally, it can be used to coat cereal, nut, fruit, and
vegetable seeds. It alters the permeability of the seed plasma membrane by raising the proline
and sugar contents.
Additionally, it increases the activity of the enzymes catalase, phenylalanine ammonia-lyase,
tyrosine ammonia-lyase, and peroxidase. Chitosan has antifungal, antiviral, and bionematicidal
properties. Chitosan facilitates the gradual release of fertilizers by acting as a carrier.
Additionally, it increases the soil’s ability to hold onto water. Additionally, chitosan is the
best chelating agent, can eliminate dyes and heavy metals, reduces algae in lakes, and acts as a
soil conditioner.

Fig 1 Application of Chitosan

Figure 2 depicts the reactive groups contained in chitosan, which include a primary amino group (C2) as well
as primary and secondary hydroxyl groups. Glycosidic linkages and the acetamide group can both be called
functional groups. These functional groups allow for numerous changes, resulting in polymers with novel
characteristics and behaviours. Chitosan derivatives have been developed with the goal of improving chitosan
qualities such as solubility and biodegradability, as well as introducing novel functionalities or properties.
Deacetylation, depolymerisation, and quaternisation, among other procedures, have all been used to improve
solubility in water-based media. New chitosan actions have been discovered following its alteration; for
example, 6-O-sulphated chitosan enhances neural development, whereas phosphorylated chitosan reduces
corrosion.

Fig. 2 Functional group of Chitosan structure that are chenucally modified.

The topic of chitosan chemistry is vast, and in this review, we will focus on two types of
processes: chitosan phosphorylation and chitosan degradation. Our group has contributed to the
synthesis of a phosphorylated derivative via a simple approach in which chitosan and
phosphorous acid are mixed at the same ratio and formaldehyde is added at 70 °C (Figure 3).

Fig. 3 Schematic diagram of phosphorylated chitosan derivative synthesis.

Functional Mechanism of Chitosan

How Chitosan Works: Unlocking Its Natural Powers

Chitosan isn’t just another biopolymer—it’s a multitasking marvel. What makes it so special? Its
unique structure, particularly those positively charged amino groups, which let it interact with
all sorts of biological molecules and surfaces. This versatility opens doors to countless
applications, from medicine to environmental cleanup.

1. Nature’s Antibiotic: How Chitosan Fights Microbes

Chitosan is a powerful germ fighter, taking on bacteria, fungi, and yeasts with a triple-threat
approach:

- Breaking Down Defenses: Its positive charge disrupts the negatively charged cell
membranes of microbes, punching holes that leak their insides out—effectively killing them.
- Starving Them Out: It grabs essential metals like calcium and magnesium that microbes need
to survive, crippling their metabolism.
- Sneak Attack: Smaller chitosan molecules can slip inside microbial cells, messing with
their DNA and stopping protein production.

2. The Invisible Shield: Chitosan’s Film-Forming Superpower

These films:
- Block unwanted gases and moisture making them perfect for keeping food fresh.
- Create a protective barrier in wound dressings, locking in moisture while keeping
infections out.

3. Sticky & Sustainable: Why Chitosan Clings and Breaks Down Safely
Chitosan plays nice with the environment—and our bodies:

- Biodegradable: Enzymes like lysozyme and chitinases break it down naturally, whether in
soil or human tissue.
- Super Sticky: Thanks to its positive charge, it bonds tightly to mucous membranes, making
it ideal for:
- Targeted drug delivery (think nasal sprays, eye drops, or gut treatments).
 - Keeping medicine in place longer where it’s needed most.
4. The Cleanup Crew: Chitosan’s Pollution-Fighting Skills
Chitosan is like a magnet for toxins:

- Traps heavy metals (lead, mercury, etc.) by forming insoluble clumps.

- Purifies water by pulling out pollutants.
- Clears up cloudy juices and beverages in food processing.

5. Healing from Within:

Chitosan’s Role in Blood
and Immunity
Chitosan doesn’t just protect—it actively helps the body heal:

- Boosts Immunity: Triggers macrophages and cytokine production, ramping up the

body’s defenses.
- Stops Bleeding Fast : Encourages platelet clumping and kickstarts blood clotting pathways.
Understanding these mechanisms helps scientists and industries harness chitosan’s full
potential—whether it’s fighting infections, packaging food sustainably, delivering drugs
more effectively, or cleaning up the environment.

Fig. 4. Functional mechanism of Chitosan

Table 1: Overview of Fungal Sources of Chitosan
Physiocochemical Chitosan Properties

1. Solubility: Chitosan is created by deacetylation of chitin, which converts certain N-

acetylglucosamine moieties into glucosamine units. The presence of a significant number of
protonated -NH2 groups on the chitosan structure contributes for its solubility in acidic
aqueous conditions, as its pKa value is roughly 6.5. Chitosan becomes soluble when
approximately 50% of its amino groups are protonated. Chitosan solubility is determined
by a variety of parameters, including polymer molecular weight, degree of acetylation, pH,
temperature, and polymer crystallinity. Homogeneous deacetylation (alkali treatment, 0 °C)
of chitin allows for the generation of polymers soluble in aqueous acetic acid solutions with
DD as low as 28%, but heterogeneous deacetylation (alkali treatment, high temperatures)
does not achieve this value. Furthermore, the samples are water soluble (DD = 49%). This
phenomenon is explained by the fact that homogenous deacetylation increases the number
of glucosamine units and modifies the polymer’s crystalline structure. Polymer DD can
result in crystal size reduction, perfection, or a new crystal structure similar to β-chitin. This
study examined the impact of crystallinity and intermolecular forces on chitosan solubility.
A parent chitosan sample was either half re-acetylated with anhydride acetic or fully N-
deacetylated under uniform conditions. After reacetylation, the polymer’s solubility was
expanded till pH 7.4, while the fully deacetylated solubility range was somewhat reduced.

The lower solubility was attributed to an increase in polymer crystallinity following

deacetylation, which counteracted the effect of an increase in glucosamine moieties. In
contrast, the half-acetylated sample showed a drop in crystallinity. The use of hydrogen
bond disruptors like urea or guanidine hydrochloride affects chitosan’s solubility window.
Broad solubility is obtained through a combination of chemical and physical breakdown of
hydrogen bonds.

2. Viscosity Polymer viscosity is an important technological parameter because extremely

viscous solutions are difficult to manage. Furthermore, viscometry is an effective instrument for
estimating the molecular weight of chitosan since it is a simple and rapid method, despite the
fact that it is not an absolute method and hence requires the measurement of solvent-specific
constants.
The Mark-Houwink-Sakurada equation connects the average molecular weight to the intrinsic
viscosity:
Η = KMvα (1)
The parameters K and α must be determined empirically. The values of K and α vary according
on solvent composition, pH, and ionic strength. Chitosan viscosity is determined by the
polymer’s molecular weight and the degree of deacetylation, and it lowers as the molecular
weight of chitosan is reduced. In fact, viscosity can be used to estimate the durability of a
polymer in solution, as it decreases during polymer storage due to degradation. Shear viscosity
increases as chitosan deacetylation progresses. The shear viscosity at the same rate was studied
in two samples with different deacetylation degrees (91% vs. 75%) and represented versus
intrinsic viscosity. It was reported that shear viscosity was larger for those samples with the
highest deacetylation degree; when the curves were evaluated, straight lines were observed in
both chitosan samples. This is explained by the nature of chitosan, which is a cationic
polyelectrolyte due to amine protonation in acidic environments. As a result, as the DD
increases, so does the chain expansion, as more glucosamine units are discovered in the polymer
chain, resulting in a higher charge density in this sample. The effect of adding isopropanol and
ethanol to a chitosan solution in 1% acetic acid on chitosan viscosity was investigated, and the
presence of the cosolvents was found to reduce the polymer’s intrinsic viscosity.

Extraction of chitosan from fungal biomass

 Conclusion
Chitosan, a naturally occurring biopolymer, offers tremendous potential in a wide range of sectors
due to its biodegradability, biocompatibility, and functional diversity. Chitosan is typically derived
from sea crustaceans, but obtaining it from soil fungi offers a sustainable, vegetarian-friendly, and
industrially viable alternative. Fungal sources not only alleviate allergenicity and seasonality
concerns, but they also produce more uniform yields and are simpler to process. This review
underlined the need of isolating soil fungal strains for chitosan production, offered detailed
extraction and characterization processes, and looked into the structural and functional properties
that make chitosan so beneficial. Its functional mechanisms, which include antibacterial action,
metal ion chelation, and film generation, make it useful in a variety of fields such as medicine,
agriculture, food preservation, and environmental cleanup.Future study should focus on optimizing
fungal cultivation and extraction processes, improving chitosan functionalization for specific
applications, and investigating genetic and biotechnological options to increase productivity and
quality. Chitosan production can be made more environmentally friendly, sustainable, and
accessible by utilizing soil fungi as a trustworthy source.

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