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Microbial Degradation of Phenol: A Review

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 Journal of Water Pollution & Purification Research
ISSN: 2394-7306 (Online)
Volume 4, Issue 1
www.stmjournals.com

Microbial Degradation of Phenol: A Review

Viraj Krishna Mishra*, and Neeraj Kumar
Department of Biotechnology, Ambala College of Engineering and Applied Research, Devsthali,
Ambala, Haryana, India

Abstract
Microorganisms have enormous potential to remove toxic organic compounds present in waste
water due to its potential to metabolize these molecules. The presence of phenol and its
derivatives is increasing in water resources due to its extensive applications. Microorganisms
have been widely utilized to reduce the level of phenol in waste water. Different bacterial and
fungal strains have been successfully studied to reduce the level of phenol in water. Both aerobic
and anaerobic degradation of phenol is reported.

Keywords: Phenol degradation, waste water, aerobic, anaerobic, microorganisms

*Author for Correspondence E-mail:viraj.krishna@gmail.com

INTRODUCTION photosynthesis of diatoms and blue green algae.

Phenol is an aromatic compound having one
hydroxyl group attached to the benzene ring.
Phenol is also known as carbolic acid, phenic
acid, phenylic acid, phenyl hydroxide or
oxybenzene [1]. It is used in the production of
slimicides, disinfectants, antiseptics, pesticides
and medicinal preparations. It is also used in the
synthesis of phenolic compounds such as,
Bisphenol A (used for producing epoxy resins
for paints, coatings and moldings, and in
polycarbonate plastic), Caprolactam (used in
the manufacture of nylon and polyamide
plastics), Phenyl amine (used as an antioxidant
for rubber manufacture, and as an intermediate
in herbicides, dyes and pigments, and
pharmaceuticals), Alkyl phenols (used in the
manufacture of surfactants, detergents and
emulsifiers, and also in insecticide and plastics
production), Cholrophenols (used in medical
antiseptics and bactericides such as TCP and
Dettol), Salicylic acid (used in the production
of aspirin and other pharmaceuticals) [2].
Phenol is a major pollutant included in the list
of United Nations Environmental Protection
Agency (EPA). It may be fatal by ingestion,
inhalation and skin absorption since it quickly
penetrates the skin, may cause severe irritation
to the eyes and respiratory tract. It is considered
to be potentially carcinogenic to humans and
may be lethal to fish at concentrations of 5–25
mg/l. A concentration of 0.1 μg/ml and higher
concentration of phenol cause the inhibition of
Phenol is inhibitory to the nitrification process
at and above 1000 mg/l. Accidental and
intentional ingestions of phenol have been
reported. As little as 50–500 mg has been fatal
in infants. Deaths in adults have resulted after
ingestions of 1–32 g of phenol. The NO2 groups
of phenol are inhibitory to methanogenesis.
Phenols and catechol reveal peroxidative
capacity, they are hematotoxic and hepatotoxic.
It provokes mutagenesis and carcinogenesis in
humans as well as other living organisms [3].
Phenol is a corrosive substance in acute
exposure; phenol denatures proteins and
generally acts as a protoplasmic poison. Phenol
may also cause peripheral nerve damage (i.e.,
demyelination of axons).
Systemic poisoning can occur after inhalation,
skin contact, eye contact, or ingestion.
Typically transient central nervous system
(CNS) excitation occurs, and then profound
CNS depression ensues rapidly. Damage to the
nervous system is the primary cause of death
from phenol poisoning. However, damage to
other organ systems (e.g., acid-base imbalance
and acute kidney failure) may complicate the
condition. Symptoms may be delayed for up to
18 h after exposure. If more than 60 square
inches of skin are affected, there is risk of
imminent death. Most hematological changes
(e.g., hemolysis, methemoglobinemia, bone
marrow suppression, and anemia) can be
detected by blood tests or simply by the color

JoWPPR (2017) 17-22 © STM Journals 2017. All Rights Reserved Page 17
Microbial Degradation of Phenol
or appearance of the blood.
Methemoglobinemia is a concern in infants up
to 1 year old. Repeated phenol exposure in the
workplace has caused renal damage including
kidney inflammation, swelling in the kidney
tubules and cells, and degenerative changes in
glomeruli. Liver damage and pigment changes
of the skin have been noted in some workers.
Chronic exposure has also been correlated with
an increased risk of coronary artery disease and
insufficient blood supply to the heart in workers
[2]. Since phenol is toxic and causes pollution,
it must be removed from the environment.
Various physicochemical processes such as
chemical oxidation, adsorption, extraction,
pervaporation etc. have been studied to reduce
the load of phenol from waste water. Chemical
oxidation is used to remove pollutants. In this
process, one or more electrons transfer from the
oxidant to the targeted pollutant. Chemical
oxidizing agent’s convert phenol in
hydroquinone and then quinone [4]. ‘’Removal
of phenol by adsorbtion is done by use of
activated carbon. The adsorption capacity of
activated carbon relies on the physical
properties of adsorbent and the solution
conditions [5]. Coal, fly ash, sludge, biomass,
zeolites, and other adsorbents are low cost
absorbents for removal of phenol and their
removal capacity is low as compared with
synthetic resin.
The process of liquid–liquid extraction is also
known as solvent extraction. It is a method that
is employed for partitioning of a solute between
two immiscible phases, i.e., typically an
organic solvent and an aqueous solution [6].
Selectively removal of waste constituent in the
waste water is achieved when it is contacted
with an organic solvent, because it is more
soluble in the solvent than in wastewater.
There are several organic solvents such as
toluene, n-hexane, cyclohexane, benzene,
ethylbenzene, cumene, acetate esters (ethyl
acetate, isopropyl acetate, n-butyl acetate, n-
pentyl-acetate, iso-pentylacetate, n-hexyl
acetate, and cyclo-hexyl acetate), di-isopropyl
ether, methyl-iso-butylketone used for
extraction of phenol from water [7, 8]. Matjie
and Engelbrecht used “Phenosovan” an
extraction process for removal of phenol from
water in gasification plants. They utilize di-
JoWPPR (2017) 17-22 © STM Journals 2017. All Rights Reserved
Mishra and Kumar
isopropyl ether (DIPE) to recover phenol [9].
Physicochemical methods have several major
drawbacks such as energy consumption, high
cost, production of hazardous byproducts, poor
efficiency and applicable only for limited
concentration range [10, 11].
In solvent extraction, major limitations are its
high cost and contamination of solvent used for
extraction in waste water stream.
In adsorption adsorbent used is of high cost and
spent adsorbent is considered as hazardous
waste. The reactors used in chemical oxidation
operate under high temperature and high
pressure rate therefore it requires huge amount
of energy [12]. Biological treatments of phenol
are very attractive and efficient methods and
overcome from major drawback mentioned in
physicochemical methods. Biological methods
use pure and mixed microbial strains that
degrade phenol and its derivatives into nontoxic
products and it is cost effective [13–15].
MICROBIAL DEGRADATION OF
PHENOL
Phenol, an aromatic hydrocarbon, is degraded
by various microorganisms which utilizes
phenol as the sole carbon source for the growth
of the organisms. Among the various
microorganisms, Pseudomonas putida is the
most popular organism for the degradation of
phenol as this species uses phenol as the carbon
source. Arinjay et al., investigated biological
degradation of phenol and catechol by P. putida
MTTC 1194 in basal salt medium in shake flask
at 29.9 ± 0.3 ºC and pH of 7.1 and found that P.
putida MTTC 1194 degraded 1000 mg/l of
phenol in 162 h and 500 mg/l of catechol in 94
h [15].
Sarwade and Gawai studied biodegradation of
phenol alkaliphillic strain Bacillus badius D1
which degraded 85% of 1.5 g/l phenol present
in the medium in 48 h at 37 ºC, pH of 9 and 110
rpm [3].
Wang et al., immobilized the Acinetobacter sp.
Strain PD 12 in PVA beads; this immobilized
cells were capable of removing 99.6% of 500
mg/l phenol and metabolizing up to 1100 mg/l
of phenol at pH 7.2 [16]. Phenol is degraded
under both aerobic and anaerobic conditions.
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Journal of Water Pollution & Purification Research
Volume 4, Issue 1
ISSN: 2394-7306 (Online)
Table 1: Phenol Degrading Microorganism [18].
Source Genus
Alcaligenes
Anthrobacter
Pseudomonas
Bacteria
Pseudomonas aeruginosa MTCC 4996
Pseudomonas aeruginosa CC7CCAB919095
Cyanobacterium
Bacillus
Candida
Fusarium
Fungi
Graphium
Ochromonas
Aspergillus
Phanerochaete
Rhodococcus erythropolis UPV -1
Rhodococus
Yeast Rhodotorula
Sphingomonas chlorophenolica RA 2
Sphigmonas
Trichosporon
Aerobic Biodegradation of Phenol
Biodegradation of phenol under aerobic
condition is initiated by oxygenation. In this
process, the aromatic ring is initially
monohydroxylated by a mono oxygenase
phenol hydroxylase at an ortho position to the
pre-existing hydroxyl group to form catechol.
This is the main intermediate resulting from
metabolism of phenol by different microbial
strains. Depending on the type of strain,
catechol is oxidized via ortho-cleavage
pathway by catechol 1,2-dioxygenase, leading
to the formation of succinyl Co-A and acetyl
Co-A or by meta-pathway to 2-
hydroxymuconic semialdehyde by catechol
2,3- dioxygenase that leads to the formation of
pyruvate and acetaldehyde [17].
Anaerobic Biodegradation of Phenol:
Anaerobic degradation of phenol is less
advanced than aerobic process. It is based on
the analogy with the anaerobic benzoate
pathway proposed for Paracoccus denitrificans.
First step in the anaerobic pathway is
carboxylation of phenol at the para position to
4- hydroxybenzoate; here the enzyme involved
is 4-hydroxy benzoate carboxylase. Anaerobic
biodegradation of phenolic compounds
JoWPPR (2017) 17-22 © STM Journals 2017. All Rights Reserved
Species
Alcaligenes faecalis
Alcaligenes xylosoxidans Y234
Arthobacter species
Arthrobacter citreus
Arthrobacter chlrophenolicus A6
Pseudomonas putida
Pseudomonas cepacia
Pseudomonas pictorum
Pseudomonas aeruginosa
Cyanobacterium synechococcus
Bacillus species strain PHN1
Bacillus brevis
Bacillus badius
Candida tropicalis
Candida tropicalis NICM 3556
Fusarium species
Graphium sp.FIB4
Ochromonas danica
Aspergillus awamori NRRL 3112
Phanerochaete chrysosporium
Rhodotorula creatinivora
Trichosporon species LE3
Trichosporon cutaneum R57
including o-cresol, catechol and ortho -
halogenated phenol is mediated via
carboxylation and followed by dihydroxylation
OPTIMIZATION OF SIGNIFICANT
FACTORS FOR BIODEGRADATION
OF PHENOL
Biodegradation of phenol rely upon various
factors such as concentration of phenol present
in growth medium for growth of phenol
degrading bacteria, nitrogen source,
temperature condition, pH, salt concentration,
buffer concentration etc. Therefore, finding
optimal condition for phenol biodegradation is
important. Optimization of microbial growth
conditions, particularly physiological and
chemical parameters (medium components) are
of primary importance in the development of
any biodegradation process.
The degradation efficiency of microbes is
maximum when the process is carried out under
optimum growth conditions. The modeling and
optimization methodologies, like one factor at a
time (OFAT) in which optimization is done
changing one parameter at a time and keeping
the others constant. This optimization study
does not consider the interaction effects among
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Microbial Degradation of Phenol
the variables as any process is influenced by
several variables. To overcome this limitations
statistical techniques such as Plackett–Burman
design (PBD), Central composite design (CCD)
and Box–Behnken Design (BBD) were used.
These methodologies are efficient and effective
approach for systematic investigation of the
target factors. PBD is an effective screening
design which considerably diminishes the
number of experiment and gives information
for the evaluation of the target factors as much
as possible.
BIODEGRADATION KINETICS OF
PHENOL
The kinetics study of phenol biodegradation is
very for efficient removal of phenol.
Understanding the kinetics of cell growth and
biodegradation of phenolic compounds is
essential for system optimization. Most of the
microorganism follow Haldane’s model of
growth kinetics; only few follow the Monod’s
and Linerized–Haldane’s model of growth
kinetics. μg is a function of S. If phenol is
considered to be noninhibitory compound and
so was represented by Monod’s noninhibitory
kinetics equation [3] as given below:
(1)
If phenol is considered to be growth inhibitory
compound then Haldane’s growth model was
selected due to its mathematical simplicity and
wide acceptance for representing the growth
kinetics of inhibitory substrates. The Haldane’s
inhibitory growth kinetics equation is as
follows:
(2)
At higher substrate concentrations, S >>Ks, the
above equation reduces to the following:
(3)
Arinjay et al., studied biodegradation kinetic of
phenol and catechol using P. putida (MTCC
1194). Biological degradation of phenol and
catechol by a bacterial strain of P. putida
(MTCC 1194) in basal salt medium (BSM) was
investigated in shake-flask experiments at
29.9±0.3 ºC and pH of approximately 7.1. The
well-acclimatized culture of P. putida (MTCC
JoWPPR (2017) 17-22 © STM Journals 2017. All Rights Reserved
Mishra and Kumar
1194) degraded the initial phenol concentration
of 1000 mg/l and initial catechol concentration
of 500 mg/l completely in 162 h and 94 h,
respectively [15]. Both the phenol and catechol
were observed to be the inhibitory compounds.
Haldane’s model for growth kinetics fit for this.
The decay coefficients have been found to be
0.0056 h−1 and 0.0067 h−1 for the growth on
phenol and catechol, respectively. Besides, the
yield coefficient for the growth on phenol and
catechol were found to be 0.65 mg/mg and 0.50
mg/mg, respectively [15].
Essam et al., isolated Alcaligenes sp. TW1 from
the activated sludge of the industrial waste
water treatment plant of a Coke company
having high phenol tolerance. Showing
Haldane kinetics model to growth on phenol as
sole carbon and energy source at 25° C and they
have obtained kinetic parameters as: µmax = 0.58
h-1, Ks= 10 mg/l, and Ki = 152–550 mg/l and
they have found biomass yield coefficient
ranged from 0.55 to 0.64 mg dry cell mass/mg
phenol for maximum 1200 mg/l phenol
concentration. Haldane model best fits to
growth data of microbes for phenol
biodegradation since phenol is a growth
limiting substrate. It is widely accepted for
representing the growth kinetics of inhibitory
compounds due to its mathematical simplicity
and accuracy. Thus, there is scope to evaluate
the growth kinetic parameters for isolated
strains using Haldane model [20-22].
Banerjee et al., studied kinetics of degradation
of phenol with pure cultures Bacillus cereus
MTCC 9817 strain AKG1 and B. cereus MTCC
9818 strain AKG2 in batch mode for phenol
concentrations in the range of 100–2000 mg/l
with an interval of 100 mg/l by applying kinetic
models such as Yano model, Haldane model,
Aiba model, Webb model, and Edward model.
It was found that the Haldane inhibitory model
fits the experimental data fairly well for both
the strains and estimated kinetic parameters
were: q=27.85 h-1, Ks= 59,150 mg/l and Ki=
2.411 mg/l [14].
CONCLUSIONS & FUTURE
PROSPECTS
Increasing pollution is posing risk to natural
resources. Presence of phenol and its aromatic
derivatives in waste water is toxic. The
exposure of these compounds to living
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Journal of Water Pollution & Purification Research
Volume 4, Issue 1
ISSN: 2394-7306 (Online)
organisms poses threat to life. Waste water
need to be sufficiently treated to reduce the
level of phenol. Along with physical and
chemical treatment process, microbial
treatment process can enhance the efficiency of
treatment process. Considering the importance
of microorganisms in the treatment of waste
water, more studies are needed to identify the
effective microorganisms and to synchronize
microbial treatment process with established
process of waste water treatments.
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Cite this Article
Mishra VK, Kumar N. Microbial
Degradation of Phenol: A Review.
Journal of Water Pollution &
Purification Research. 2017; 4(1):
17–22p.
Page 22