Improving

Biodegradation Potential
of Dioxin-Degrading
Bacteria
Polychlorinated Polycyclic
Aromatics

Dioxins
polychlorinated dibenzo-
P-dioxins

and
dibenzofurans
Dioxins is one of the most toxic
chemicals known and very stable
in the environment.

Therefore, the removal of dioxins
in river, sea and all public water
bodies is very important and
urgent matter.
Dibenzo-p-dioxins
are generated as by-products
in the manufacturing of
herbicides,
insecticides,
fungicides,
paper pulp bleaching, and
in incineration.
Dibenzo-p-dioxin and its
chlorinated relatives are
chemically stable and
accumulate in milk and
throughout the food chain,
creating significant health
concern.
Dioxin can be degraded in
water to the detection limit
using a photochemical
degradation process by a
combination of ozone and
ultra violet (UV) light
Photochemical Degradation
Photochemical Degradation
Dibenzofuran is released to
the environment in
atmospheric emissions
involved with the combustion
of coal, biomass, refuse, and
diesel fuel.
Wastewater emissions can
occur from coal tar, coal
gasification, and shale oil
operations.
If released to the atmosphere,
dibenzofuran will exist primarily in the
gas-phase where it will degrade relatively
rapidly by reaction with photochemically
produced hydroxyl radicals (estimated
half-life of 11.3 hr in average air).

A small percentage of the dibenzofuran
released to air will exist in the particulate
phase which may be relatively persistent
to atmospheric degradation.
A process that uses naturally
occurring or genetically
engineered microorganisms
such as yeast, fungi, and
bacteria to transfer harmful
substance into less toxic or non
toxic compounds
Bioremediation
BIODEGRADATIVE ECOSYSTEM

BIODEGRADATION PROCESS
ENVIRONMENTAL INFLUENCES

PHYSICAL CHEMICAL BIOLOGICAL

- TEMPERATURE INORGANIC PROPER MICROBIAL
- SOLUBILITY ORGANIC POPULATION
- PRESSURE INHIBITORS - CROSS FEEDING
- WATER AVAILABILITY - SYMBIOTIC
- SURFACE EFFECT - COMPETITION
Entry into Bacterial Cell
Dibenzo-p-dioxin
degradation
pathway
Growth
and
Reproduction
Catalyzed by Enzymes
CELL
Basic Metabolism Process of Bacteria
ENERGY
SOURCE
NUTRIENTS
NEW
CELL
MASS
H
2
O
CO
2

Carbon
source
Schematic Diagram of Biodegradation
Oil
Microbe
CO
2
+H
2
O
CO
2
+H
2
O
2
.
Microorganisms eat
oil and other
organic
contaminants.
Microbes digest
oil and convert it
to CO
2
and H
2
0
Microorganisms
release CO
2 ,
H
2
0
and metabolites
1.
3
.
Sphingomonas
Scientific classification

Kingdom: Bacteria
Phylum: Proteobacteria
Class: Alpha Proteobacteria
Order: Sphingomonadales
Family: Sphingomonadaceae
Genus: Sphingomonas
Sphingomonas
Gram-negative, rod-shaped,
chemoheterotrophic, strictly aerobic
possess ubiquinone 10 as the major
respiratory quinone,
contain glycosphingolipids (GSLs)
instead of lipopolysaccharide in their
cell envelopes, and typically produce
yellow-pigmented colonies.
Sphingomonas
were subdivided into four genera:

Sphingomonas,
Sphingobium,
Novosphingobium and
Sphingopyxis.
sphingomonads
widely distributed in nature,
Isolated from many different
land and water habitats,
as well as from
plant root systems,
clinical specimens,
and other sources
Due to their biodegradative and
biosynthetic capabilities,
sphingomonads have been utilized
for a wide range of biotechnological
applications, from bioremediation
of environmental contaminants to
production of extracellular
polymers such as sphingans used
extensively in the food and other
industries.
Sphingomonas adhaesiva
Sphingomonas aerolata
Sphingomonas aquatilis
Sphingomonas asaccharolytica
Sphingomonas aurantiaca
Sphingomonas baekryungensis
Sphingomonas chungbukensis
Sphingomonas cloacae
Sphingomonas echinoides
Sphingomonas elodea
Sphingomonas faeni
Sphingomonas koreensis
Sphingomonas mali
Sphingomonas melonis
Sphingomonas oligophenolica
Sphingomonas parapaucimobilis
Sphingomonas paucimobilis
Sphingomonas phyllosphaerae
Sphingomonas pituitosa
Sphingomonas pruni
Sphingomonas roseiflava
Sphingomonas sanguinis
Sphingomonas suberifaciens
Sphingomonas taejonensis
Sphingomonas trueperi
Sphingomonas ursincola
Sphingomonas wittichii
Sphingomonas xenophaga
Sphingomonas yabuuchiae
Species
Sphingomonas sp. A1 have got
Superchannels
Can take up macromolecules
like alginate and degrade by
alginate lyase
Pit in the outer membrane
Alginate binding proteins in
periplasm
ATP-binding cassette (ABC)
transporter
Spinnomonas strain A1
Can not degrade Dioxins
Plasmid pBE11

Plasmid pBE11
Alginate import genes
aly, ccpA, algS, algM1,
algM2, algQ1, algQ2, al-IV
Time course of Bacterial cell
growth (solid line) and
dibenzofuran concentration
(dashed line) grown in W medium
() with 0.5% alginate () 10 mM
dibenzofuran () or both ()
incubated at 30
o
C, 160 rpm for 4 d.
RW1 (pBE11)
RW1 (pKS13)
Spinnomonas
wittichii RW1 strain
Spinnomonas
wittichii RW1
(pKS13)
Spinnomonas wittichii
RW1 (pBE11) strain
Can degrade Dioxins Only slight growth
on dibenzofuran
Plasmid pBE11
Utilized 1, 10 and 100
mM dibenzofuran in
presence of alginate 0.5%
Utilized 1, 10 and 100 mM
dibenzofuran in presence of
alginate 0.5%
Showed lag time for
dibenzofuran utilization
Without Aliginate
Gene
No lag period
Longer cultivation time
4d
Shorter cultivation time
RESULTS
No growth in the
absence of alginate
Utilized 100mM dibenzofuran
in absence of alginate 0.5%
Very slow growth on
Dibenzo-p-dioxin
Could also degrade Dibenzo-
p-dioxin rapidly
Alginate import genes are
active in the absence of
alginate also.
alginate import is constitutive
No pit structure No pit structure Pit Structure observed
Spinnomonas
wittichii RW1
strain
Spinnomonas
wittichii RW1
(pKS13)
Spinnomonas wittichii
RW1 (pBE11) strain
Cell surface structure of RW1 or
RW1 (pKS13) White bar shows 0.1 m
Cell surface structure of RW1 (pBE11)
White bar shows 0.1 m
Conclusions
The superchannel promotes
dibenzofuran import in RW1 (pBE11)
Although there is no structural
relationship between dibenzofuran
and alginate, the bacteria reacted
similarly.
RW1 (pBE11) was effective in
environmental remediation of
dibenzofuran.
Improved bacterial remediation of
organic pollutants can be achieved
by increased uptake capabilities of
target organisms by incorporating
non specific superchannel.
This method has the potential of
improving the capacity of microbial
degraders suitable for
bioremediation of various pollutants.