Pollutants
Most organic and inorganic chemicals are subject
to enzymatic attack through the activities of living
organisms
Type of pollutants
BTEX (Benzene, Toluene, Ethylbenzene, Xylene)
PAH (Polycyclic aromatic hydrocarbon)
Nitroaromatic compounds
PCB (Polychlorinated biphenyls)
Chlorinated aliphatic compounds like, trichloroethylene (TCE), dichloroethylene (DCE), vinyl
chloride (VC)
MTBE: Methyl tert-butyl ether
Agricultural chemical wastes (Triazine, DDT)
Compounds with differential aerobic vs. anaerobic
degradation potential
Benzene, polynuclear aromatic
hydrocarbons (PAH)
Stable ring structures, aerobes
utilize oxidizing power of
oxygenases to initiate degradation
Anaerobes lack similarly powerful
oxidants
Compounds are readily degraded
aerobically, but persistent in
anaerobic environments
Degradation of petroleum in marine environments
Alkane Degradative Pathways
The degradation of n alkanes
of medium chain length by
Pseudomonas putida is
initiated by alkane
hydroxylase.
➢ Terminal oxidation of n-alkanes. α-and ω-hydroxylation is catalyzed by the same set of enzymes.
➢ With bacteria, steps 1, 2 and 3 are catalyzed by alkane hydroxylase, fatty alcohol dehydrogenase and fatty
aldehyde dehydrogenase, respectively.
➢ With yeast (Candida maltose), step 1 is catalyzed by P450 monooxygenase, while steps 2 and 3 are catalyzed
either by fatty alcohol oxidase and fatty aldehyde dehydrogenase, respectively.
Yeast (Candida
maltose)
Proposed scheme of the n-alkane degradation pathway and its subcellular localization in C. maltosa including the newly found role
of P450 in the oxygenation cascade from n-alkanes to ,-dioic acids. ER, endoplasmic reticulum; FAOD, fatty alcohol oxidase;
ALDH, fatty aldehyde dehydrogenase.
Degradation of cyclohexane
➢ Cycloalkanes, including condensed cycloalkanes, are degraded by a co-oxidation mechanism. The formation
of a cyclic alcohol and a ketone has been observed. A monooxygenase introduces oxygen into the cyclic
ketone and the cyclic ring is cleaved.
Degradation of Toluene
Divergent Pathways for the Aerobic Degradation of Toluene
The pathways from the top to the bottom are found with Pseudomonas putida (TOL), Pseudomonas putida F1, Pseudomonas
mendocina KR1, Pseudomonas pickettii PKO1, and Bukholderia cepacia G4, respectively.
Degradation of fatty acid Coenzyme A
(- oxidation procedure)
➢ In this process, fatty acid molecules are
broken down in the cytosol in prokaryotes
and in the mitochondria in eukaryotes to
generate acetyl-CoA.
➢ It is referred to as beta-oxidation because FAD: Flavin adenine dinucleotide
the beta carbon of the fatty acid undergoes
oxidation to a carbonyl group. NAD: nicotinamide adenine
dinucleotide
Activation of fatty acids
[Link] the cytosol of the cell, long-chain fatty acids are activated by ATP and
coenzyme A, and fatty acyl-CoA is formed.
[Link] ATP is converted to AMP and pyrophosphate (PPi), which is cleaved by
pyrophosphatase to two inorganic phosphates (2 Pi). Because two high-energy
phosphate bonds are cleaved, the equivalent of two molecules of ATP is used for
fatty acid activation.
➢ FAD accepts hydrogens from a fatty acyl-CoA in the first step. A double bond is produced between
the α- and β-carbons, and an enoyl-CoA is formed. The FADH2 that is produced interacts with the
electron transport chain, generating ATP.
➢ Enzyme: Acyl-CoA dehydrogenase (Multiple variants of this enzyme)
➢ H2O adds across the double bond, and a β-hydroxyacyl-CoA is formed.
➢ Enzyme: Enoyl-CoA hydratase
➢ β -Hydroxyacyl-CoA is oxidized by NAD+ to a β-ketoacyl-CoA. The NADH that is produced
interacts with the electron transport chain, generating ATP.
➢ Enzyme: L-3-hydroxyacyl-CoA dehydrogenase (which is specific for the L-isomer of the β-
hydroxyacyl-CoA).
➢ The bond between the alpha and beta carbons of the β-ketoacyl-CoA is cleaved by a thiolase that
requires coenzyme A. Acetyl-CoA is produced from the two carbons at the carboxyl end of the
original fatty acyl-CoA, and the remaining carbons form a fatty acyl-CoA that is two carbons
shorter than the original.
➢ Enzyme: β –ketothiolase
➢ The shortened fatty acyl-CoA repeats these four steps. Repetitions continue until all the carbons of
the original fatty acyl-CoA are converted to acetyl-CoA.
Enzymes for -oxidation
•Acyl CoA dehydrogenase: Forms a double bond between the α and β
carbon atoms in the fatty acid chain. Produces one FADH2.
•Enoyl CoA hydratase: Incorporates a water molecule into the fatty
acid chain, thereby breaking the double bond between the α and β
carbon atoms.
•3-Hydroxy-acyl CoA dehydrogenase: Dehydrogenates the fatty acid
chain again, thereby forming a double bond between the β carbon
and the oxygen molecule. Produces one NADPH.
•Acyl CoA acyltransferase: Cleaves acetyl CoA off the end of the fatty
acid chain with the addition of CoA to the β carbon.
Oxidation of odd-chain fatty acids
Odd-chain fatty acids produce acetyl-CoA and propionyl-CoA.
➢ These fatty acids repeat the four steps of the β-oxidation spiral,
producing acetyl-CoA until the last cleavage when the three
remaining carbons are released as propionyl-CoA.
➢ Propionyl-CoA, can be converted to succinyl-CoA.
Oxidatively degraded to CO2 and H2O
provide the energy needed
to drive microbial metabolic processes
Krebs Cycle
It is an eight-step process. Krebs cycle or
TCA cycle takes place in the matrix of
mitochondria under aerobic conditions.
GDP: Guanosine di-phosphate
Surfactants: structural diversity: difficult to formulate
Guidelines for biodegradation and the need to ``acclimate'' the bacteria population
Synthetic surfactants
1. Highly branched detergents degrade slowly in part.
2. Consequently, surfactants with branched chains that are shorter than those with linear chains containing the same
number of carbon atoms degrade more slowly. The possible reason may be the ease of binding of the surfactant to a
degradative enzyme.
3. The slower rate of degradation of surfactants in concentrated sol. than in dilute sol. suggests that minor
constituents of the surfactant mixture inhibit the microbial degradative enzymes.
These guidelines also apply to the alkyl substituent of the alkyl- and alkyl- phenol ethoxylates
1. The alkyl chain will degrade slowly if it is highly branched, and then the ethoxylate grouping will degrade.
2. The rate of degradation of the alkyl chain decreases as the number of ethoxylate groups increases.
The extent of degradation of the ethoxylate chain of an alkylphenol ethoxylate decreases as the number of ethoxylate
groupings increases from 5 to 20.
As it increase the difficulty of transport of the more hydrophilic detergent (with more ethoxylate groupings) through the
hydrophobic cell membrane of microorganisms.
3. The alkylphenol ethoxylates degrade more slowly than the alcohol ethoxylates sometimes the partially degraded
compound ppt out at waste disposal and its anaerobic degradation product (Nonlyphenol) is toxic.
Environmental degradation of polymers
Proteins: polymers of amino acids
Starch and Cellulose: polymers of sugars
Natural rubber is not easily biodegradable
➢ Globally, only 18% of plastics waste are recycled, and
24% are incinerated
➢ 58% is placed in landfills (dumping yards)
➢ Relatively little decomposition takes place in landfills, so
very little of this plastic material disappears with time
(stable for more than 500 years)
Photochemical decomposition Triggered by Carbonyl Groups
absorb UV light at 300-325 nm and initiate reactions leading to the cleavage of the polymer backbone
Polymers have been prepared in which carbonyl groups were purposely incorporated into the chain to facilitate their
extensive photochemical degradation.
v Poly- styrene cups in which 1% of the styrene units also contained a ketone grouping broke down to a wettable
powder after standing outside in the sun for 3 weeks
Biodegradation
Most of the biodegradable polymers contain functional groups that are subject to attack by microbial enzymes and
contain ester or amide groups that can be hydrolyzed or linear chains that can be oxidatively cleaved
Problem: inability of microorganisms to ingest high polymers through their cell walls 500 Da.
v Some microorganisms that degrade polyesters secrete esterases, enzymes that catalyze the hydrolysis of ester
groups, which break the polymer into smaller fragments that can then be ingested.
Some typical polymers that are biodegradable under aerobic conditions
Stockholm Convention In 2001
POP: Persistent Organic Pollutant
➢ remain intact for exceptionally long periods of time
(many years),
➢ become widely distributed throughout the
environment as a result of natural processes
involving soil, water, and, most notably, air;
➢ accumulate in living organisms including humans,
and are found at higher concentrations at higher
levels in the food chain
➢ And are toxic to both humans and wildlife.