C H A P T E R

9
Fungi as potential candidates for
bioremediation
Rajesh Kumar Singh1, Ruchita Tripathi1, Amit Ranjan2,
Akhileshwar Kumar Srivastava3
1
Department of Dravyaguna, Institute of Medical Sciences, Banaras Hindu University,
Varanasi, India; 2Department of Kayachikitsa Institute of Medical Sciences, Banaras Hindu
University, Varanasi, India; 3The National Institute for Biotechnology in the Negev,
Ben-Gurion University of the Negev, Beer-Sheva, Israel

1. Introduction
The condition of environment directly influences the quality of life in the ecosystem on
earth. Human beings are most dominant animal on the earth, have developed the science
and technology for their comforts, and industrialized at large scale for the fulfillment of their
requirements, but unfortunately these produce adverse affect on environment, often resulting
to the extinction of several species of the earth. The civilization and industrialization produce
large amount of organic and inorganic pollutants, as well as nuclear wastes, which are
generally flushed into the river or dumped into the soil. The flushing and dumping of
such materials pollute to system, creating a serious problem for the survival of organisms.
Such pollutants are disposed by making a hole into land and filling it by waste material or
just filling the low-land areas just away from the cities, but continuous increases of the
pollutants need new place in near future, which develops other issues such as lack of water-
bodies, floods, etc. These methods of disposal are difficult to sustain because of limited space
and are also not economic and healthy (Karigar and Rao, 2011). Hence, bioremediation is
alternative, economical, and acceptable method to dispose these polluting materials.
Bioremediation is a process in which the waste and hazardous materials are transformed
into nonhazardous or less-hazardous substances by microbes such as bacteria, fungi, algae,
etc. The applications of fungal microbes in bioremediation have been well-known as fungal
bioremediation or mycoremediation. The fungi decompose the biomass residues and
chemical pollutants by producing several enzymes such as amylase, protease, lipase, nuclease,

Abatement of Environmental Pollutants

https://doi.org/10.1016/B978-0-12-818095-2.00009-6 177 Copyright © 2020 Elsevier Inc. All rights reserved.
178 9. Fungi as potential candidates for bioremediation

etc., which degrade the cell walls, proteins, lipids, DNA, and other organic materials. These
microbes are used for biotransformation of organic waste materials and removal of pollutants
from environment (Balabanova et al., 2018). Fungi are capable to degrade the organic
chemicals, metals, metalloids, and radionuclides, by secreting enzymes and other chemicals
through chemical modification and/or influencing chemical bioavailability. It has also adopted
to survive in diverse condition continuously. Fungi are competent for metabolize various
environmental chemicals and utilize its product for survival without any additional need of
nutrition. It degrades pollutants through several extracellular oxidoreductases for decompos-
ing the lignocelluloses along with several pollutants. Fungi metabolize and immobilize the
metals, metalloids, and radionuclides in the mycosphere, store in various parts of the cell, or
translocate through fungal hyphae. It also degrades the compounds that are not intoxicated
by bacteria efficiently such as dioxins and 2,4,6-trinitrotoluene, drugs, etc. These features enable
fungi as potential candidate for removing organic or metal contaminants in the soils, water,
and air and can be used for bioremediation with cost-effective and nature friendly ways
(Harms et al., 2011). This chapter explains the diverse role of fungi group in overcoming the
pollutant toxicity through a robust technique bioremediation as summarized in Fig. 9.1.

1.1 Fungal enzymes for bioremediation

The enzymes of many fungi have low specificity, which enables the fungal strain to metab-
olize several compounds of different pollutants even dissimilar in structure simultaneously,
like Phanerochaete chrysosporium is a crust fungi, which degrades many harmful chemicals
including benzene, ethylbenzene, xylene, toluene, organochlorines, N-heterocyclic explosives,

FIGURE 9.1 Describing role of fungi in bioremediation and involvement of fungal enzymes in removal/
transformation of materials.
 1. Introduction 179
polycyclic aromatic hydrocarbons (PAHs), nitroaromatic compounds, pesticides, synthetic
dyes, polychlorinated dibenzo-p-dioxins, and synthetic polymers simultaneously even in
mixture of all (Kues, 2015). The major groups of enzymes involved in the degradation of wastes
materials and pollutants include extracellular oxidoreductases, cell-bound enzymes, and other
transferase enzymes.

1.1.1 Extracellular oxidoreductases

The several extracellular oxidoreductase enzymes are secreted by fungi to degrade the
organic compounds such as laccases, tyrosinases, lignin peroxidases, manganese peroxidases,
etc. (Harms et al., 2011). These enzymes enabled the fungal strains to grow on recalcitrant
substrates such as lignocelluloses that are not efficiently degraded by most bacteria, and fungi
could survive on the substrates where bacteria cannot grow. These properties of fungi are
advantageous over bacteria for application in bioremediation (Verma et al., 2017).
The prominent copper-containing laccases are mainly produced in Basidiomycetes and
Ascomycetes and used for decolorization and detoxification in textile industries, bleaching
of ink in paper industries, degradation and detoxification of recalcitrant compounds in waste-
water, hazardous compounds arising from coal processing, and degradation of soil pollutants
(Alneyadi et al., 2018; Spina et al., 2015).
Another copper-containing enzyme tyrosinase also oxidizes the phenols and chlorinated
phenolic compounds. It has potential for the removal of such chlorinated compounds from
the environment. Presently, many peroxidases are characterized in microbes especially
from fungi, oxidize pollutants, and convert them into nonhazardous products with high
redox potentials (Al-Maqdi et al., 2017).

1.2 Cell-bound enzymes

Many compounds of waste materials and pollutants are permeable to the fungal cells
membranes, which are chemically catabolized through several intracellular fungal enzymes
such as polyaromatic hydrocarbons and dioxins. Cytochrome P450s also have a vital role
in intracellular degradation of the pollutant and involve in fungal metabolism of many drugs
such as antiinflammatory drugs, lipid regulator drugs, antiepileptic and antianalgesic drugs,
and diphenyl ether herbicides. The pollutants under fungal bioremediation undergo degrada-
tion by fungal metabolism till they are mineralized (Harms et al., 2011). The metabolic
activities may further also involve in formation of metabolites at different stages of oxidation
in the degradation process.

1.3 Transferases
Transferases include mainly aromatic nitroreductases, quinone reductases, etc., which
transfer the functional groups and convert the hazardous pollutants into nonhazardous
products. The transferase enzymes degrade the pollutants containing hydroxyl groups by
converting it into the conjugates. The conjugates are stored, fixed, or secreted into environment
in inactive conjugated forms (Morel et al., 2013). The different enzymes and components are
involved in remediation of contaminated materials as mentioned in Table 9.1.
TABLE 9.1 The details of enzymes producing fungal taxa and mechanism of action.

180
S.No. Enzymes Fungal taxa Enzyme occurrence Mechanism of action References

1 Laccases Ascomycota and Extracellular O2-dependent one-electron oxidation of (Baldrian, n.d.; Majeau
Basidiomycota organic compounds. et al., 2010)
2 Tyrosinases Ascomycota, Intracellular (but O2-dependent hydroxylation of monophenols (Halaouli et al., n.d.;
Basidiomycota, and sometimes to o-diphenols (cresolase activity) Ullrich and Hofrichter,
Mucoromycotina extracellular) Oxidation of o-diphenols to catechols 2007)
(catecholase activity).

3 Lignin Basidiomycota Extracellular H2O2-dependent one-electron oxidation of (Hofrichter et al., 2010;

peroxidases aromatic compounds. Karich et al., 2017; Ruiz-
Dueñas et al., 2009)

9. Fungi as potential candidates for bioremediation

4 Manganese Basidiomycota Extracellular H2O2-dependent one-electron oxidation of (Hofrichter et al., 2010,
peroxidases Mn2þ to Mn3þ, which subsequently oxidizes n.d.; Ruiz-Dueñas et al.,
organic compounds. 2009)
5 Versatile Basidiomycota Extracellular H2O2-dependent direct one-electron oxidation (Hofrichter et al., 2010;
peroxidases of aromatic compounds Martínez, 2002; Ruiz-
H2O2-dependent one-electron oxidation of Dueñas et al., 2009)
Mn2þ to Mn3þ, which subsequently oxidizes
organic compounds.
6 Coprinopsis cinerea Basidiomycota Extracellular H2O2-dependent one-electron oxidation of (Hofrichter et al., 2010;
peroxidase aromatic compounds. Ikehata et al., 2005)
7 Dye-decolorizing Basidiomycota Extracellular H2O2-dependent one-electron oxidation of Hofrichter et al. (2010)
peroxidases organic compounds
Additional hydrolyzing activity.
8 Caldariomyces Ascomycota Extracellular H2O2-dependent halogenation of organic Hofrichter et al. (2010)
fumago haeme compounds in the presence of halides
thiolate (one-electron transfer), H2O2-dependent
chloroperoxidase one-electron oxidations of phenols and anilines
in the absence of halides, and H2O2-dependent
peroxygenation (two-electron oxidation),
leading to epoxidation of (cyclo)alkenes,
hydroxylation of benzylic carbon and
sulphoxidation of S-containing organic
compounds.
9 Haemethiolate Basidiomycota Extracellular H2O2-dependent peroxygenation of aromatic, Hofrichter et al. (2010)
peroxygenases aliphatic, and heterocyclic compounds, leading
to aromatic and alkylic carbon hydroxylation,
double-bond epoxidation, ether cleavage,
sulphoxidation or N-oxidation reactions
(depending on the substrate), H2O2-dependent
one-electron abstractions from phenols, and
H2O2-dependent bromination of organic
substrates.
10 Cytochrome P450 Ascomycota, Cell bound Incorporation of a single atom from O2 into a (Kasai et al., 2010;
monooxygenases Basidiomycota, substrate molecule, with concomitant Subramanian and Yadav,
Mucoromycotina, reduction of the other atom to H2O. 2009; Ullrich and
and Hofrichter, 2007; Yadav
Chytridiomycota et al., 2006)
11 Phenol 2- Ascomycota and Cell bound Incorporation of a single atom from O2 into a (Hofrichter et al., n.d.;
monooxygenases Basidiomycota substrate molecule, with concomitant Ullrich and Hofrichter,
reduction of the other atom to H2O. 2007)

1. Introduction
12 Nitroreductases Ascomycota, Cell bound NADPH-dependent reduction of (Bhushan et al., 2002;
Basidiomycota, and nitroaromatics to hydroxylamino and Crocker et al., 2006;
Mucoromycotina amino(nitro) compounds and of nitro Esteve-Núñez et al., 2001;
functional groups of N-containing Fournier et al., 2004;
heterocycles. Scheibner et al., 1997)
13 Reductive Basidiomycota and Cell bound Two-component system comprising a (Jensen et al., 2001;
dehalogenases perhaps membrane-bound glutathione S-transferase Nakamiya et al., 2005)
Ascomycota that produces glutathionyl conjugates with
concomitant chlorine removal and a soluble
glutathione conjugate reductase that releases
reductively dechlorinated compounds.
14 Miscellaneous Ascomycota, Cell bound Formation of glucoside, glucuronide, xyloside, Kasai et al. (2010)
transferases Basidiomycota, and sulfate, or methyl conjugates from
Mucoromycotina hydroxylated compounds.

181
182 9. Fungi as potential candidates for bioremediation

2. Fungal bioremediation
2.1 Toxic recalcitrant compound
The poor electron donor compounds such as polychlorinated dibenzo-p-dioxins (PCDDs)
and polychlorinated dibenzofurans (PCDFs) occurring at very low concentration are oxidized
to the electronegativity of the chlorine atoms by accepting the electrons from the aromatic
rings (Hammel, 1995). The compounds with more than one chlorine atom are not employed
as a source of carbon and energy by bacteria. Therefore, the biotransformation of the toxic
compounds, PCDD 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), through aerobic
and anaerobic bacteria is slow process ranging to months; however, it is not completely
biotransformed and finally halted at the stage of less chlorinated aromatic products. Compar-
atively, the white-rot fungus Phanerochaete sordid converts the compound 2,3,7,8-TCDD into
chlorocatechols within 10 days and other fungus of same species P. chrysosporium mineralizes
the intermediate products. The fungus P. sordida has no specific to particular compound and
it transforms compounds containing different chlorine atoms, namely PCDDs (6e8 chlorines)
and PCDFs (4e8 chlorines) (Lipnick et al., 2000). The ascomycete Cordyceps sinensis has great
potential to remove the highly chlorinated dioxins at fast rate similar to white-rot fungi. The
2,4,6-trinitrotoluene (TNT) is also highly oxidized to the electronegativity of its nitro groups.
The transformation of TNT is accomplished mostly through aerobic bacteria into
monoamino-dinitrotoluenes, diamino-nitrotoluenes, and hydroxylamino-dinitrotoluenes that
acquire to produce mutagenic azoxy-tetranitrotoluenes, although the transformation of TNT
through anaerobic bacteria mostly halts at the stage of triaminotoluene, whereas white-rot
and litter-decaying basidiomycetes mineralize TNT rapidly (Esteve-Núñez et al., 2001).
Researches into use of nonspecific enzymes through biotechnological approaches must eluci-
date the problems generated during the reactions. The specific fungal transformation products
of PAHs have less concentration of mutagenic and carcinogenic metabolites in which most
transformed products are lower toxic than its parent compounds. The formations of PCDDs
and PCDFs on enzymatic oxidative coupling of chlorophenols are required to assess the
structure, biological activity, stability, and environmental behavior (Lamar et al., 2003). The
laccase-catalyzed transformation of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydro-
chloride (EDCs) to oligomeric coupling products reduces the endocrine-disrupting activity of
such compounds. Similarly, the toxicity is reduced by laccases and other phenoloxidases for
covalently linked chlorophenols, other phenols, and TNT metabolites to components of soil
humic substances.
PAHs as good substrates in respect to energetic perspective and its low molecular mass
PAHs are commonly used by bacteria. The bioavailability decreases inverse to molecular
mass, only a few bacteria grow at high molecular mass PAHs with five or more aromatic
rings. The fungal species of Ascomycota, Basidiomycota, and Mucoromycotina having a
nonspecific mechanism to detoxify the compounds hydroxylate to PAHs intracellularly
and then transfer to water-soluble compounds that could be excreted out. In addition,
different fungi utilize the extracellular oxidoreductases in PAH degradation and mineralize
to high molecular mass PAHs, such as highly carcinogenic benzo[a]pyrene and others
(Mao and Guan, 2016).
 2. Fungal bioremediation 183

2.2 Heavy metal

Fungi for remediation of materials (metals, metalloids, and radionuclides) are identified on
the basis of morphological, ecological, and biochemical properties. The accumulations of toxic
metals in environment are accomplished through different stages of the life cycle of metallic
substances, from the mining of metal ores to the final disposal of metal-containing hazardous
substances. The mobilization of metals from geogenic sources, agriculture field, and incident
like fires are attributed a constant input into waters and the atmosphere and from it further is
translocated into soils and aquatic sediments (Sanyal et al., 2005). The contaminated environ-
ment with metals could be hazardous for mainly to near of ore mines and smelters or at
disposal sites where proper remediation is not adequate. The main risk of metallic products
is entirely associated with the nondegradable element. So, biological methods are not capable
to reduce metal toxicity in an irreversible way. Such rising risk from such elements could be
overcome through biological conversion into less toxic species or by separation from the
affected biota. With advancement of technology, it has also drawn much interest toward
elimination of metals from wastewater streams for restoration of valuable metals. It accom-
plishes through such methods such as mobilization and immobilization in the mycosphere,
sorption to cell walls and uptake into fungal cells (Ayangbenro and Babalola, 2017). The
metals could be chemically converted after incorporation into fungus and accumulated in
different parts of the cell or translocated from fungal hyphae into symbiotic plant.

2.3 Municipal solid waste

Nowadays, the municipal solid waste (MSW) in developing countries has been one of the
major problems for environment along with public health. For removal of MSW, the process
incineration and landfilling are mainly employed, although incineration is a very costly
process, hence the landfilling sites produce the secondary environmental pollution such as
fouling of air, bad odor, and increased pathogenic activity in soil. In another way, the coun-
tries with limited land areas are not fit for composting and landfilling; hence it is essential to
expand the land areas. Deshmukh et al. have explained about several modern technologies
for resolving of such misplaced resources. It has also explored the composting and bio-
methanation method through anaerobic process to manage the MSW with production of
valuable substances such as volatile fatty acids (VFAs), biogas, and organic residue/compost
for application as fertilizers (Deshmukh et al., 2016). For increasing the potential and rates of
these methods, the fungi and its hydrolytic enzymes could be used for the transformation of
complex polymeric compounds of MSW to simple one acting as mediators for VFA and
biogas production. The enzymes cellulolytic, hemicellulolytic, pectinolytic, and amylolytic
of Aspergillus niger are used to increase the efficiency of steam, acid, and base pretreated
kitchen waste residues for solid-state fermentation of enzymes (Rani et al., 2014). The using
of these enzymes in pretreatment enhance the potential for hydrolysis and saccharification
of certain biomasses such as willow and rice straw, by application of a fungal consortium
containing two fungi Armilleria gemina and Pholiota adipose. The white-rot fungi also increase
the efficiency for composition of other residual biomass by utilizing the used biomass in soil
application. The accumulated copper in wood could be remediated through highly efficient
184 9. Fungi as potential candidates for bioremediation

wood-rotting fungi Antrodia xanthan and Fomitopsis palustris (Deshmukh et al., 2016;
Voberkova et al., 2017). Trichoderma viride is one of the promising fungi for conversion of
organic municipal solid waste to harmless substances. The main factors such as pH and
temperature are good indicator for the end of the bioconversion of municipal solid waste
(Gautam et al., 2012).

3. Fungi in bioremediation
Fungi could survive in different habitats that colonize in soil matrix along with freshwater
and also in marine habitats. Fungi also grow in different climatic conditions including the
stress condition and inseminate the spores through air that aid in balancing of ecosystem.
It has been reported that the fungi can also survive in effluent treatment plants (ETPs) during
treatment of wastewaters. The surviving ability in diverse habitats and potential for produc-
ing the diverse enzymes indicates fungi as potential candidates for bioremediation.

3.1 White-rot fungi

White-rot fungi are used for biodegradation of lignininous substances in atmosphere that
attribute in the carbon recycling. The accumulation of endocrine disrupting chemicals (EDCs)
and TrOCs such as pharmaceuticals and personal care products (PPCPs) develop acute and
chronic toxicity to aquatic organisms and also harmful for human health have been remedi-
ated by white-rot fungi. Many previous studies explained the efficiency of white-rot fungi for
bioremediation, namely Phanerochaete chysosporium, Bjerkandera adjusta, and Pleurotus sp.,
which secrete the varieties of ligninolytic enzymes, e.g., laccases and peroxidases (Voberkova
et al., 2017; Yang et al., 2013). The ligninolytic enzymes of white-rot fungi have been used for
conversion of different organic pollutants such as pesticides of the contaminated wastewaters
through enhancing the microbial activity using a biopurification system (BPS). The deposited
lignin granules in the lignocellulosic fibers are separated by applying the pressure refining
that has been enhanced through using of ligninolytic enzymes of white-rot fungus Ceriporiop-
sis subvermispora exhibited greater delignification in comparison to pressure refined Miscan-
thus than milled Miscanthus. Extracellular ligninolytic enzymes of Agaricomycete and
white-rot from Amazon forest fungi are also capable for adsorption, degradation or decolou-
rization of dyes such as Direct Blue 14 manufactured of Pleurotus and Remazol Brilliant
Blue-R. The fungal groups such as Coriolus versicolor, Inonotus hispidus, and Phlebia tremellosa
have capacity to decolorize of dye effluent, although 38 species of white-rot fungi have been
explained in reduction of total phenolics (>60%) and color (70%) from olive-mill waste-
water. The remediation of cresolate in the contaminated soil has been accomplished through
two strains of white-rot fungi: T. versicolor and Lentinus tigrinus. The cresolate-polluted soil is
generally mixture of contaminants such as residual petroleum hydrocarbons and high
molecular weight PAH fraction after a biopiling treatment. The potential degradation of
the contaminants could be held through biostimulation of lignocellulosic substrate with bio-
augmentation of fungi (Yadav et al., 2006; Yang et al., 2013). However, such type of treatment
 3. Fungi in bioremediation 185
possibly chances to promote the growth of neighbor microbes that may influence to the
augmented organism, hence before applying such treatment in filed it is required to validate
the study at a small scale. Moreover, other properties of white-rot fungi secreting a laccase
enzyme are also applied for degradation of substituted organic. Looking to such important
features in bioremediation, it has been approached by escalating the laccase production in
white-rot fungi, T. versicolor and P. ostreatus, through solid state fermentation on orange peels
followed by further investigating of its efficiency for bioremediation of PAHs such as
phenanthrene and pyrene. The production of laccase (3000 U/L) from T. versicolor cultures
and P. ostreatus produced 2700 U/L laccase exhibited better removal of phenanthrene and
pyrene. However, more study at the molecular level to understand the potential role of fungi
in bioremediation is required.

3.2 Marine fungi

The marine fungi produce the efficient secondary metabolites, biosurfactants, enzymes,
polysaccharides and fatty acids that are used in bioremediation of hydrocarbons and heavy
metals. The adaptation of marine fungi to high saline conditions and pH makes more biolog-
ical advantageous in extreme condition than the terrestrial fungi. The potential of marine
microbes for remediating the metal ion indicate to the promising nature of extremophilic
microbes for bioremediation and in nanotechnology. Thatoi et al. have explained the diverse
potential applications of marine fungi from mangrove with emphasizing on its diversity,
promising ecological role, and biotechnological application for source of drugs, enzymes,
biodiesel, biopesticides, and bioremediation (Thatoi et al., 2013). Recently, it has been eluci-
dated the potential role of the derived enzymes from marine fungi and its biotechnological
application (Bonugli-Santos et al., 2015). Marine fungi are highly tolerable to high concentra-
tions of heavy metals and its interaction to metal ions of marine ecosystems could be employed
for preparation of metal nanoparticles. Fungi have capability to synthesize nanoparticles both
extra- and intracellularly that are attributed for applications in different areas from textile
industries, food preservations, to medicines and clinical microbiology, etc.
The various factors of fungi have been explored for increasing the efficiency of bioremedi-
ation of toxic and persistent organic pollutants. The production of laccase enzyme from
marine fungi is highly tolerable to salinity and phenolics have been implied in the isolated
Trichoderma viride Pers NFCCI-2745 of an estuary polluted with phenolics (Divya et al.,
2014). The enzyme mediated bioremediation has also been explained for decolorizing
Remazol Brilliant Blue-R dye applying basidiomycetes of marine sponges, and anthraqui-
none dye Reactive Blue 4 by C. unicolor, a marine white-rot basidiomycete. Gao et al. have
reported that the biostimulation and bioaugmentation have adverse affect to the transforma-
tion of persistent organic pollutants (POPs) such as PCB 118 by marine fungi belonging to
genus Penicillium containing maifanite. Another POP such as pentachlorophenol has been
transformed even at high concentrations through marine-derived fungus, Trichoderma
harzianum and other marine fungi such as Mucor, Aspergillus, and slime mold exhibited
efficient bioremediation for water-soluble crude oil fractions (0.01e0.25 mg/mL) at higher
level causing toxicity to the organisms (Gao et al., 2013).
186 9. Fungi as potential candidates for bioremediation

3.3 Extremophilic fungi

The extremophilic fungi are very imperative for industries producing extremophilic
enzymes posses several unique properties such as thermo tolerance, pH tolerance, and sustain
to other stress conditions. The effluent treatment plant has strong niche that is targeted for
fungi with different ways of bioremediation applications at high levels of pollutants from
industrial effluents.
The extremophilic fungi with such properties are better candidates for economical level
and eco-friendly along with bio-transformation of raw materials, i.e., in food industries,
textiles, and animal fodder preparation, etc. The implications of metallophilic microbes in
bioremediation produce problems with heavy metals from the environment that is generated
by formation of nanoparticle for efficient bioremediation (Sinha et al., 2011). A psychrophilic
fungus, Cryptococcus sp. occurring in deep-sea sediments have capability to tolerate at high
amount of heavy metals (100 mg/L) ZnSO4, CuSO4, Pb(CH3COO)2 and CdCl2. Several
hydrolytic enzymes also exhibit activity in extremophilic conditions and participate in
remediation methods under stress conditions such as high salinity and extra-heavy crude oil
(ECHO) contamination of oil belts. The laccases activity under extreme environment are
used in bioremediation processes in Pestalotiopsis palmarum in presence of wheat bran and
lignin peroxidases are produced from extra heavy crude oil as a source for carbon and energy
(Majeau et al., 2010). The enzymes such as chitinases from psychrophilic fungus, Lecanicillium
muscarium, are used to enhance the activity of insecticides. The isolation of extremophilic fungi
for bioremediation processes from extreme environments like deep biosphere habitat having
fumarolic ice caves on Antartica’s Mt. Erebus are also used to characterize the other unique
fungi using as energy sources to remediate the human contamination in such extreme places.

3.4 Symbiotic association of fungi with plants and bacteria

Fungi have close relation with plants and bacteria to reduce the hurdle of restricted growth
under different environmental conditions. Arbuscular mycorrhizal fungi (AMF) are common
symbiotic association with fungi and higher plants, there fungal partner involves in removal
of pollutant with larger surface area by absorbing the contaminants through its hyphae and
spores. AMF colonized in root of plants are used as phytoremediation for polluted ground-
water with several hazardous substances in a constructed wetland. Some plant-associated
fungi, e.g., A. nidulans, Funalia trogii, Irpex lacteus, P. ostreatus, etc., are capable to survive
in and decolorize textile industry effluents. The colonized AM fungus Rhizophagus custos in
root organ are able to survive in high levels of PAHs mainly anthracene with less production
of toxic from anthraquinone. The ectomycorrhizal fungi, Suillus bovinus and Rhizopogon
roseolus associated with Pinus involve in removal of cadmium that is also subjected to the ef-
fect of other environmental factors such as types of nutrients and pH (Mao and Guan, 2016).
Another implication of such fungi is to overcome the technical rifts of algal bio-fuels and
photosynthetic biorefineries through co-cultivation of microalgae and fungi for removal of
single algal cells from fermentation medium. This allows their extraction and harvest through
simple filtration that increases the yielding of biomass, lipid, and bioproduct. Instead of
co-culture studies for bioremediation, a keen understanding of its application associated with
the interaction between multitudes of metabolic pathways of different organisms is required.
 4. Technology advancement 187

4. Technology advancement
Bioremediation of toxic substances is one of reliable methods for cleanup of polluted sites
and fungi play important role in remediation of hazardous substances containing different
constituents in ecosystem with multiple modes for overcoming the problems of contamina-
tion. Nonetheless, their implications are mainly associated with environmental factors and
long lag phase, sludge generation, and difficult methods, which may have direct impact
on application of fungal biomass in bioremediation. Many advance technologies have been
developed in field of fungal bioremediation to reduce the associated problems. One such
advance processes associated with enzymes of the fungal biomass reduces bioremediation
time, no lag phase, minimal sludge generation, and easy process control. Though enzymes
create itself other problems of high cost and less shelf life due to lower stability, although
enzymes production in whole cell and its immobilization have been enhanced their stability
by increasing shelf life and achieved for reusing the enzyme with affordable costs. To date,
the various bioreactors; fluidized beds and rotating biological contactors have been developed
for remediating of contaminants with fungi (Gautam et al., 2012). However, novel bioreactor
systems are being constructed for elimination of dyes, e.g., Reactive Green 19 by white-rot
fungi (Sari et al., 2016). A two-stage reactor have been succeed in remediation of azo dye
Reactive Blue 222 along with Photo-Fenton’s and aerobic treatment through two white-rot
fungi P. ostreatus IBL-02 and P. chrysosporium IBL-03 (Kiran et al., 2013). A white-rot fungus,
T. versicolor, revealed potential removal of TrOCs through fungal membrane bioreactor in
non-sterile condition at 2 days hydraulic retention time (HRT) (Yang et al., 2013). Bhattacharya
et al. have developed a novel approach for degradation of HMW-PAHs using white-rot fungus
P. chrysosporium. The remediation of benzo[a]pyrene under ligninolytic culture medium in-
creases the PAH oxidizing monooxygeneses with formation of P450-hydroxylated metabolite
that is eliminated through non-ligninolytic phase (Bhattacharya et al., 2014). The other
significant strategy associated with biopurification systems in stimulating bioremediation of
pesticides having wastewaters through highly active biological mixture of particular white-
rot fungi has been explained by Rodrı’guez-Rodrıguez et al. (Rodríguez-Rodríguez et al.,
n.d.). The sustainability and eco-friendly nature of bioremediation has been displayed for
the remediation of sewage sludge containing filamentous inoculum in a large-scale bioreactor
by implying a continuous process (Rahman et al., 2014). In spite of the fungi alone, its
co-cultures with bacteria playing as a synergistic degradation system containing Fusarium
sp. PY3, Bacillus sp. PY1, and Sphingomonas sp. PY2 remove pyrene by 96.0% and volatilized
arsenic by 84.1% (Liu et al., 2013). Other significant and innovative approach for remediation
of PAHs have been initiated by Cobas et al. (2013) in which 90% phenanthrene has been
removed within 14 days by formation of permeable novel reactive biobarriers of Trichoderma
longibrachiatum on nylon sponge. Fungal biocatalysis is employed in all cell systems associated
with textile wastewater treatment (Spina et al., 2015).

4.1 Conclusions and future prospective

The unique lifestyle and biochemical properties of fungi elucidate its importance in
biotechnological implications to transfer the hazardous substances to nonhazardous. It is a
highly demand, sustainable, cheap, and tailor-made technology developed using fungi to
188 9. Fungi as potential candidates for bioremediation

translate powerful ecosystem services into ecology-based technologies. The remediation of

contaminated land is a passive process for monitoring the natural attenuation, and such mea-
sures require more time than active ex situ remediation, so it is relied on the autochthonous
microbial communities. The low efficiency of mechanical process in spontaneous attenuation
of soil facilitates toward the establishment of filamentous fungi. Most fungi produce diverse
exudates, which serve as carbon sources for bacteria involving in remediation of pollutants.
The intrusion of plants along with mycorrhizal or other autochthonous fungi could be
promising approach for increasing the efficiency of rhizosphere bioremediation of the trap-
ped pollutants and organic contaminants. Unfortunately, relative contributions of fungi to
degradation and detoxification processes have seldom been quantified, and the same is
true for the positive effects of fungi on bacterial remediation processes.
Because of lack of knowledge of methodologies and ecological approaches, it is important to
sustain sufficient fungal biomass, activity, and enzyme secretion at pollutant site, thereby
causing inhibition of mycoremediation. Moreover, more studies on bacteriaefungi interactions
are required to understand such interactions for development of novel, ecologically stable
bioremediation approaches. The strong isotope techniques analyze a better appreciation of
fungal components in purification of nature, i.e., the fungal removal of air-borne organic
contaminants attributing into plant litter or humus that provides a relevant reservoir and
secondary source in the global cycling of these compounds. Fungi as potential driver of passive
remediation technologies with low cost could be widely accepted for risk-based cleanup stan-
dards that have been recently comprehended in the legislation of countries: the United States
and the United Kingdom. This chapter suggests that the fungi as potential approaches with
important financial, ecological, and legal reasons could be used in bioremediation of hazardous
materials.

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