Microbiol. Biotechnol. Lett.

(2019), 47(2), 278–288

http://dx.doi.org/10.4014/mbl.1903.03006
pISSN 1598-642X eISSN 2234-7305
Microbiology and Biotechnology Letters

De-novo Hybrid Protein Design for Biodegradation of

Organophosphate Pesticides
Garima Awasthi, Ruchi Yadav, and Prachi Srivastava*
Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow-226028, U.P., India

Received: March 11, 2019 / Revised: April 3, 2019 / Accepted: April 12, 2019

In the present investigation, we attempted to design a protocol to develop a hybrid protein with better
bioremediation capacity. Using in silico approaches, a Hybrid Open Reading Frame (Hybrid ORF) is devel-
oped targeting the genes of microorganisms known for degradation of organophosphates. Out of 21 genes
identified through BLAST search, 8 structurally similar genes (opdA, opd, opaA, pte RO, pdeA, parC, mpd
and phnE) involved in biodegradation were screened. Gene conservational analysis categorizes these
organophosphates degrading 8 genes into 4 super families i.e., Metallo-dependent hydrolases, Lactamase B,
MPP and TM_PBP2 superfamily. Hybrid protein structure was modeled using multi-template homology
modeling (3S07_A; 99%, 1P9E_A; 98%, 2ZO9_B; 33%, 2DXL_A; 33%) by Schrödinger software suit version
10.4.018. Structural verification of protein models was done using Ramachandran plot, it was showing
96.0% residue in the favored region, which was verified using RAMPAGE. The phosphotriesterase protein
was showing the highest structural similarity with hybrid protein having raw score 984. The 5 binding sites
of hybrid protein were identified through binding site prediction. The docking study shows that hybrid
protein potentially interacts with 10 different organophosphates. The study results indicate that the
hybrid protein designed has the capability of degrading a wide range of organophosphate compounds.

Keywords: Homology modeling, organophosphate degradation, hybrid ORF, bioremediation

Introduction degradation is not so effective, due to the toxicity of mix-

Organophosphates (OP) are highly toxic pesticides
and insecticides, used in different practices such as agri-
culture, gardens, and veterinary practices. The toxicity
of these compounds affects environment, ecosystem and
health of living beings. Biodegradation is found to be the
most authentic and applicable technique used for such
compounds. Many microorganisms are the established
sources of organophosphate degradation. It is well
reported that mixed microbial population can be more
effective than single micro-organism for the degradation
of wide range of toxic compounds [1, 2]. Sometimes bio-
*Corresponding author
Tel: +91-9453141916, Fax: +91-522-2814080
E-mail: psrivastava@amity.edu
© 2019, The Korean Society for Microbiology and Biotechnology
ture of organic pollutants. This limitation leads to
develop concepts and protocols for genetically engi-
neered microorganisms (GEMs) for biodegradation of
wide range of toxic compounds, as they contain artifi-
cially designed catabolic pathways [3].
GEMs have been reported as a potential source for bio-
degradation, which enhance degrading capabilities of a
wide range of chemical contaminants [4]. It has been
reported that the genetically engineered bacteria of
Escherichia coli and Pseudomonas putida strains were
stable and effective for the treatment of waste water [5].
The shotgun method was used clone methyl parathion
hydrolase encoding gene (mpd) with cognate regulator of
a methyl parathion (MP)- degrading strain Pseudomo-
nas putida DLL-1. Recombinant plasmid pBBR- mpd
was produced by using broad-host vector pBBR1MCS-2.

http://dx.doi.org/10.4014/mbl.1903.03006
 In Silico Designing of Hybrid Protein for Degradation of Organophosphate Compounds
This recombinant plasmid pBBR- mpd was transformed
in Sphingomonas sp. CDS-1 and finally CDS-pBBR-
mpd recombinant produced. Sphingomonas sp. DS-1
was recombinant strain for degradation of Organophos-
phate and carbamates. This recombinant organism was
having 7 times higher capacity of degrading methyl
parathione in comparison to wild type strain [6]. The
genetically engineered strain Escherichia coli JM109
(pGEX-AZR) have been reported for decolorization of
C.I. Direct Blue 71 [7]. In silico study has been carried
out for the designing and construction of metal binding
hybrid proteins expressed in Escherichia coli for the
removal of cadmium [8]. Similarly, few more research
have been carried out regarding the construction of
microbial consortia for the degradation of toxic com-
pounds such as plastic garbage [9], diesel oil [10] and
various xenobiotics compounds [11].
The studies of toxic compounds degrading proteins
have been carried out for predicting the possibilities of
biodegradation [12−16]. This study is focused to develop
a protocol for the construction of hybrid protein for bio-
degradation of wide range of organophosphates. The
hybrid protein will be designed from constructed micro-
bial gene consortia using homology modeling method.
Materials and Methods
The extensive literature search and data mining
(NCBI genomic databank) were done to collect the iden-
tified genes involved in degradation of organophos-
phates. The information gathered was in the form of
FASTA sequence and functional protein.
A BLAST search was done to identify similar genes.
Total 21 organophosphates degrading genes were identi-
fied in different strains of the microorganism. Eight dif-
ferent genes (opdA, opd, opaA, pteRO, pdeA, parC, mpd,
and phnE) were selected.
Superfamily (Conserved regions) of these genes was
identified using BLASTp and ScanProsite tool. Func-
tional classification of organophosphates degrading
genes was done on the basis of motif and its function.
Homology modeling of the hybrid sequence was done
using a multi-template method by Schrödinger software
suit [17−19]. The structure similarity of modeled hybrid
protein was analyzed by using FATCAT (Flexible struc-
ture Alignment by Chaining Aligned fragment pairs
279
allowing Twists) pair wise alignment tool [20].
Hybrid protein- ligand interaction study was done
using docking by Schrödinger software suit. The struc-
ture of organophosphate compound was searched from
the PubChem database (https://pubchem.ncbi.nlm.nih.gov/),
and the 2D chemical structure of these compounds was
downloaded and saved in SDF file. Library of these
organophosphate compounds was used for docking study
with the hybrid protein by glide dock method. The dock-
ing step includes ligand preparation, protein prepara-
tion and binding site prediction by site map tool, grid
generation, and glide dock. The Glide dock method gen-
erated pose viewer file, and this file was used to study
glide score. Protein-ligand interaction was visualized
using ligplot tool of Schrödinger software [21−25]. The
protein-ligand interaction map was used to study the
bond and other interactions between organophosphates
and hybrid protein.
Results and Discussion
Blast result of organophosphate degrading gene
Organophosphate degrading gene Opd A was identi-
fied in Agrobacterium tumefaciens [26], Opd in Flavo-
bacterium sp. [27], Pseudomonas putida, Brevundimonas
diminuta, Sphingobium fuliginis, Chryseobacterium bal-
ustinum, PTE-RO in Brevundimonas diminuta [28],
opaA in Alteromonas sp. [29], Mpd in Achromobacter sp.
mp-2 [30], Plesiomonas sp. M6 and Ochrobactrum sp.
Yw28, pdeA in Delftia acidovorans [31], parC in Pseudo-
monas aeruginosa [32], and phn E in Escherichia coli
K-12 [33] by using BLAST tool. The functional protein
identified were parathion hydrolase, phosphotriesterase,
organophosphate pesticide hydrolase and partition pro-
tein C. The 8 different genes were screened and selected
for the construction of consortium, from the identified 21
organophosphate degrading genes, as detail mentioned
in Table 1.
BLASTp result of organophosphate degrading gene
The second screening is based on different superfami-
lies. The superfamilies were identified for screened 8
genes, using BLASTp tool. The gene conservational
analysis shows that four superfamilies of organophas-
phate degrading genes identified are Metallo-dependent
hydrolases, Lactamase B, MPP (Metallophosphatase)
June 2019 | Vol. 47 | No. 2
280 Awasthi et al.

Table 1. List of Genes degrading Organophosphate and its superfamilies.

S. no Gene Protein Organisms Superfamily Accession no.
1. Opd A Phosphotriesterase A. tumefaciens Metallo-dependent hydrolases superfamily AY043245
(R. radiobacter)
2. Opd Parathion hydrolase Flavobacterium sp. Metallo-dependent hydrolases superfamily M29593.1
3. PTE-RO Phosphotriesterase variant B. diminuta Metallo-dependent hydrolases superfamily KJ680379
PTE-R0
4. OpaA Organophosphorus acid Alteromonas sp. Lactamase B superfamily U29240.1
anhydrolase-2
5. Mpd Organophosphate pesticide Achromobacter sp. mp-2 Lactamase B superfamily AY627034.1
hydrolase
6. PdeA Phosphodiesterase D. acidovorans MPP-superfamily superfamily AF548455.1
7. ParC Partition protein C P. aeruginosa No conserved domain have been detected KR106190
till yet
8. Phn E Organophosphate pesticide E. coli K-12 TM_PBP2 superfamily EG11283
hydrolase

and TM_PBP2 (Transmembrane subunit (TM) found in
Periplasmic Binding Protein (PBP)). Sequences of these
superfamilies were used for designing of hybrid ORF as
shown in Fig. 1. Finally, length of ORFs, start position
and stop position of ORFs of conserved region of selected
superfamilies were identified as details mentioned in
Table 2. The hybrid ORF gene sequence was constructed
using identified conserved region of superfamilies and
length was 3731 bp.
Template identification for homology modeling
The multi template homology modeling by Schrödinger
software suit was done to cover the maximum hybrid
ORF sequence for designing hybrid protein structure.
Four templates were selected from the blastp result of
hybrid ORF sequence and the finally selected templates
were 3S07_A, 1P9E_A, 2ZO9_B, and 2DXL_A, which
were showing similarity with the hybrid ORF sequence.
The identity% of 3S07_A and 1P9E_A is 99% and 98%
respectively whereas the same of 2ZO9_B, and 2DXL_A
is 33%. The query position of template 3S07_A and
1P9E_A are from 32 to 360 and from 649 to 979 respec-
tively, where as the query cover of both the template is
26%. Similarly, the query position of 2ZO9_B and
2DXL_A are same from 391 to 601 and query cover of
both the template is 17%. The template 3S07_A belongs
to phosphotriesterase family which is a binuclear metal-
loenzyme and 1P9E_A belong to Metallo-beta-lactamase
family. The 2ZO9_B and 2DXL_A belong to Calcineurin-
like Phosphoesterase family. These can hydrolyze
organophosphate pesticides and nerve agents, methyl
parathion and phosphoesterase and also capable of
nucleotide binding respectively. All these proteins of the
selected templates are also capable of metal binding.

Homology modeling multi template method by using

Schrödinger software suit
The hybrid protein structure was built by using the
selected four templates as shown in Fig. 2(A) and the
structural prediction was done through Ramachandran
plot [34−37] using Rampage [38] shown in Fig. 2(B). This
result shows stability of hybrid protein, where 96.0%
Fig. 1. Designing of Hybrid ORF degrading organophos- residues in most favored region, 4.0% in allowed region,
phates. and 0% in outlier region, which was above the expected

http://dx.doi.org/10.4014/mbl.1903.03006
 In Silico Designing of Hybrid Protein for Degradation of Organophosphate Compounds 281

Table 2. Detail of selected four conserved superfamilies and their function.

ORF start ORF stop Length of
S. no Superfamily Sequence Function
position position ORF
1 Metallo- GFTLTHEHICGSSAGFLRAWPEFFGSRKALAEKAVRGLRH 49 357 312 Phosphotriesterase
dependent ARSAGVQTIVDVSTFDIGRDVRLLAEVSRAADVHIVAATG
hydrolases LWFDPFYPLSMRMRSVEELTQFFLREIQHGIEDTGIRAGIIK
superfamily VATTGKAITPFQELVLKAAARASLATGVPVTTHTSASQRD
GEQQAAIFESEGLSPSRVCIGHSDDTDDLSYLTGLAARGY
LVGLDRMPYSALGLEGNASALALFGTRSWQTRALLIKALI
DRGYKDRILVSHDWLFGFSSYVTNIMDVMDRINPDGMA
FVPLRVIPFLREKGVPPETLAGVTVANPARFL
2 Lactamase B YLVNTGSKLVLVDTGAAGLFGPTLGRLAANLKAAGYQPE 100 302 203 organophosphate
superfamily QVDEIYITHMHPDHVGGLMVGEQLAFPNAVVRADQKE pesticide hydrolase
ADFWLSQTNLDKAPDDESKGFFKGAMASLNPYVKAGK
FKPFSGNTDLVPGIKALASHGHTPGHTTYVVESQGQKLA
LLGDLILVAAVQFDDPSVTIQLVSDSKSAAVERKKAF-
ADAAKGGYLIAAAH
3 MPP- FIHITDIHLVEQGRGALYGHDPGKRFERCIDSVIAEHADAA 4 237 236 Phosphodiesterase
superfamily SCVITGDLAHVGHPDAYRQLSEQCARLPMPVHLILGNHD
SRTNFRERFPQVPVDSNGFVQYEQAIGRFRGLFLDTNEP
GTHCGVFCEQRANWLSQRLAEADDSPVLLFMHHPAFH
LGIPVMDRIGLVDNEWLLTALKGHEHRVKHLFFGHIHRPI
SGSWRGIPFSTLRGTNHQVALHLRESEDIPGSFEPPQYAV
4 TM_PBP2 MAVTLQIAVWGTALAVVLSIPFGLMSAENLVPWWVYQP 69 247 188 organophosphate
superfamily VRRLMDACRAINEMVFAMLFVVAVGLGPGWGILPGLGF pesticide hydrolase
AGVLALFIHTTGVLSKLLSEAVEAIEPGPVEGIRATGANKLE
EILYGVLPQVMPLLISYSLYRFESNVRSATVVGMVGAGGIG
VTLWEAIRGFQFQQTCALMVLIIVTVSLL

90% in the most favored regions. It is expected to have a (favored region) and outer limit (allowed region). The
good quality model, if the most favored regions are over generously allowed regions are located as pale greenish
90%. This Ramachandran plot shows secondary struc- yellowish area. The disallowed regions (white area) gen-
ture like α helix and β sheet in the fully allowed part erally involve steric hindrance between the side chains

Fig. 2. (A) Structure of Hybrid ORF protein designed using multi template homology modeling by Schrödinger software suit.
(B) Ramachandran plot of hybrid protein structure designed, where 96.0% residues in favored region, 4.0% in allowed
region and 0% in outlier region.

June 2019 | Vol. 47 | No. 2

282 Awasthi et al.

Table 3. Comparative study of hybrid protein versus known protein.

UniProt Entry Structure Raw Equivalent
S. no Protein Microorganism PDB ID P-value RMSD gaps
ID similarity score positions
1 Phosphotriesterase Agrobacterium tumefaciens Q93LD7 3SO7 significantly 984 0.00E+00 329 0.00 0
similar (0.00%)
2 Parathion hydrolase Flavobacterium sp. P0A433 1P6C significantly 958.13 0.00E+00 327 0.39 0
similar (0.00%)
3 Phosphotriesterase Brevundimonas diminuta A0A060GYS1 4PCP significantly 955.29 0.00E+00 327 0.46 0
variant PTE-R0 similar (0.00%)
4 Organophosphate Achromobacter sp. mp-2 Q693X3 3T1G significantly 328.72 1.32E-03 246 2.92 82
pesticide hydrolase similar (25.00%)
5 Phosphodiesterase Delftia acidovorans Q8GMW5 1NNW NOT 172.56 1.69e-01 178 3.85 151
significantly similar (45.90%)
6 partition protein C Pseudomonas aeruginosa A0A0H4P9E0 No structure was identified

of one amino acid with the backbone of the succeeding
amino acid. The glycine is an exception, since it lacks the
side chain responsible for the clash and can adopt phi
and psi angles in disallowed region of the Ramachan-
dran plot, which is acceptable [39]. The multi template
homology modeling was done to cover the whole hybrid
ORF sequence for the designing hybrid protein structure.
residue, the active binding sites for molecules. The
Aspartic acid was major identified amino acid on all the
five binding sites. The other identified amino acids were
Phenylalanine (9), Leucine (8), Arginine (8), Glycine (7),
Alanine (6), Histidine (5), Serine (5) and Valine (5). The
binding sites help in the interaction of protein with the
ligand.

Comparative study of designed hybrid protein with

known protein
The comparative study of designed hybrid protein was
done with the screened proteins mentioned in Table 1.
To compare the structural similarity of hybrid protein
with the selected known protein was done using FATCAT
pair wise alignment tool. The hybrid protein was show-
ing structural similarity with the screened protein phos-
photriesterase, parathion hydrolase, phosphotriesterase
variant PTE-R0 and organophosphate pesticide hydro-
lase. Phosphodiesterase was showing no similarity with
hybrid protein. No structure was identified for Partition
protein C. The raw score, p value, equivalent position,
RMSD and gaps were analyzed as mentioned in Table 3.
The phosphotriesterase has shown the maximum simi-
larity and raw score 984, with hybrid protein.

Binding site of hybrid protein

The binding sites of hybrid protein are predicted to
increase the docking efficiency. The five binding sites
were identified in structure of hybrid ORF protein,
which can be visualized in Fig. 3. The details of binding
sites are mentioned in Table 4. The highest site score
Fig. 3. Binding sites of hybrid ORF protein by schrödinger
was 1.024 of site 1, having maximum sites and amino software suit, (A) site 1, (B) site 3, (C) site 2, (D) site 4, (E) site
acid residues. The other four sites also have amino acid 5.

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 In Silico Designing of Hybrid Protein for Degradation of Organophosphate Compounds 283

Table 4. Details of identified binding sites of hybrid ORF protein.

Binding
S. Site D Contact Hydro- Hydro- Don/ Fig.
site Size Volume Exposure Enclosure Balance Residues Name
No. score score potential phobic philic acc 13
name
1 Site 1 1.024 106 0.97 316.246 0.584 0.733 0.911 0.328 1.259 0.261 1.189 Chain A:, His 200, Thr 201, Leu (a)
270, Ser 202, Phe 271, Phe
305, Ala 203, Gly 272, Ser 204,
His 229, Ser 307, Arg 274, Tyr
308, Ile 105, Arg 253, Arg 279,
Tyr 256, His 54, Asp 232, His
56, Trp 130, Phe 131, Asp 132,
Gly 59, Ala 269, Lys 168, Asp
300, Leu 302
2 Site 3 0.883 64 0.604 122.108 0.492 0.742 1.03 0.128 1.821 0.07 0.493 Chain A: Arg 66, Phe 103, Glu (b)
158, Asp 159, Cyc 58, Gly 106,
Arg 107, Asp 108, Val 109, Thr
160, Ser 60, Ser 61, Ala 62, Gly
63, Arg 110, Phe 64, Leu 65
3 Site 2 0.78 55 0.691 127.939 0.601 0.653 0.873 0.251 1.221 0.206 0.914 Chain A: Ala 77, Asn 311, Leu (c)
78, Ile 312, Met 313, Phe 303,
Asp 314, Gly 304, Phe 305,
Asp 317, Val 309, Lys 81, Phe
71, Phe 72, Thr 310
4 Site 4 0.703 36 0.56 93.296 0.625 0.692 0.955 0.196 1.287 0.152 0.907 Chain A: His 88, Asp 322, Ala (d)
89, Trp 301, Gly 323, Leu 302,
Met 313, Met 324, Phe 303,
Ala 325, Asp 317, Lys 81, Arg
84, Gly 85, Asn 320, Pro 321
5 Site 5 0.606 22 0.505 96.04 0.732 0.692 0.983 0.218 1.077 0.203 1.566 Chain A: Val 124, Asp 34, Leu (e)
35, Leu 357, Val 36, Asn 37,
Pro 359, Val 195, Arg 163, Ile
153, Ala 164, Gly 165

Protein-Ligand interaction study using docking
To check the efficiency of hybrid protein, its ligand
interaction studies were done by docking study using
Schrödinger software suite. The docking study helps in
assessment of the binding affinity of ligand and protein.
The hybrid ORF protein was constructed for the degra-
dation of organophosphate compounds; therefore, vari-
ous organophosphate compounds were used as a Ligand.
The most commonly known ten organophosphate com-
pounds were selected randomly as a ligand for docking
from the literature. These organophosphate compounds
were Acephate, Parathion-methyl, Fonofos, Chloropyri-
fos-methyl, Diazinon, Bromofos, Chlorphoxim, Monocro-
tophos, Chlorpyrifos and Malathion. These selected
organophosphate compounds are used as insecticide and
acaricide, they are also cholinesterase inhibitor. Acephate,
Parathion-methyl and Chlorpyrifos are white crystalline
solid compound; Fonofos is a light-yellow liquid with a
pungent mercaptan-like odor used in cultivation of corn
plant. Diazinon is widely used as pesticide for rice, fruit,
trees and ornamental plants. Chlorpyrifos methyl,
Monocrotophos and Malathion widely used against pests
on cotton, sugar cane, tobacco, potatoes, peanuts, toma-
toes, and ornamentals plants. Malathion is a wide spec-
trum aliphatic organophosphate insecticide. Malathion,
Diazinon and Chlorpyrifos are widely used for both
domestic and commercial agricultural purposes. These
are used to kill mosquitos, cockroaches, fleas, termites,
pet flea, tick collars, ticks on cattle and Mediterranean
fruit flies. These are highly toxic compounds; weather
inhaled, ingested or absorbed by skin such as Chlor-
phoxim is toxic when it comes in contact with skin, Bro-
mophos cross the placental barrier. Malathion is toxic to
aquatic organisms but less toxic for birds and mammals.
Human exposed to high levels Chlorpyrifos has autoim-
mune antibodies, which is common in autoimmune dis-
orders also associated with higher risks of lung cancer
[40, 41].
The docking studies were showing the interaction of
ligand with the hybrid ORF protein. The highest 10 mol-

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284 Awasthi et al.

Table 5. Details of organophosphate compounds and modeled ORF protein interaction using docking.
S. No Compound name Pubchem ID Glide score Protein-ligand interaction

1. Acephate 319061378 -5.99

2. Parathion-methyl 249980167 -5.2

3. Fonofos 319071828 -5.13

4. Chlorpyrifos-methyl 319077673 -5.04

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 In Silico Designing of Hybrid Protein for Degradation of Organophosphate Compounds 285

Table 5. Continued.
S. No Compound name Pubchem ID Glide score Protein-ligand interaction

5. Diazinon 319556886 -4.78

6. Bromofos 250121756 -4.67

7. Chlorphoxim 319334751 -4.63

8. Monocrotophos 319302319 -4.51

June 2019 | Vol. 47 | No. 2

286 Awasthi et al.
Table 5. Continued.
S. No Compound name Pubchem ID Glide score
9. Chloropyrifos 319061835
10. Malathion 319556698
ecules, which shows the maximum docking score were
acephate (-5.987), parathion-methyl (-5.2), fonofos
(-5.125), chlorpyrifos (--5.043), diazinon (--4.775), chlor-
phoxim (-4.625), monocrotophos (-4.509), chloropyrifos
(-4.491) and malathion (-4.491). These glide score -5.987
to -4.491 are showing effective interaction between
ligand and protein. The details of the protein- Ligand
interaction map are shown in Table 5.
The interaction of hybrid protein with organophos-
phates was due to the hydrogen bonds (side chain and
back bone) and Pi-Pi staking. The arginine (253), trypto-
phan (130) and histidine (His 200) were showing interac-
tion via hydrogen bond with the ligand. The hydrogen
(back bone) bond interaction was observed in monocroto-
phos with Serine (H→Ser307). The hydrogen bond (side
http://dx.doi.org/10.4014/mbl.1903.03006
Protein-ligand interaction
-4.49
-4.01
chain) was observed in acephate (Arg→O), chlorpyrifos-
methyl (Arg→O), chlorphoxim (Arg→N and Arg→O),
chloropyrifos (Arg→N) and diazinon (Arg→N) with argi-
nine. Similarly, interaction was also observed in mala-
thion with tryptophan (Trp→O) and histidine (His→O).
The pi-pi staking interaction mainly occurs with tyro-
sine (Tyr 308), tryptophan (Try 130), phenylalanine (Phe
131) and histidine (His 200). The pi-pi staking was
observed in chlorphoxim, fonofos and parathion-methyl
with tyrosine (Tyr 308), in parathion-methyl with tryp-
tophan (Try 130) and in bromofos, chloropyrifos and
chlorphoxim with phenylalanine (Phe 131) amino acid.
All the organophosphates were showing solvent expo-
sure on groups like -Br, -Cl, -C and -O- attached at side
chain of aromatic rings. All the interactions were in the
 In Silico Designing of Hybrid Protein for Degradation of Organophosphate Compounds
vicinity of hydrophobic zone and salt bridge zone.
Potentiality of work signifies that the in silico
designed hybrid protein has a potent interaction with
organophosphate compounds. These docking results
indicate that the hybrid protein will be capable of
degrading wide range of organophosphate compounds. It
has been reported by the previous researchers that the
interaction of in silico constructed genetically engi-
neered microorganism with the target compounds to
study the biodegradation can be successfully performed
by docking analysis. This in silico analysis will optimize
and enhance the bioremediation process before wet labo-
ratory experiments [42, 43]. The in silico designed
hybrid protein model has shown high interaction with a
wide range of organophosphates and was verified by
molecule docking. This in silico study developed a proto-
col for the construction and analysis of the genetically
engineered microorganisms. This study will help in min-
imizing the wet laboratory efforts and results obtained
will support to development GEMs in vitro. This hybrid
protein consortium will be a landmark for fighting
against the problem of organophosphate-induced pollu-
tion and may give a solution to such kind of many more
issues.
Acknowledgments
We would like to express our sincere gratitude to Dr A B Pant, Princi-
pal Scientist, System Toxicology and Health Risk Assessment Group,
CSIR-Indian Institute of Toxicology Research, Lucknow, India, for pro-
viding all necessary help and scientific suggestions during this
research work. I would like to acknowledge Bioinformatics analysis
tools and data bases present in Amity Institute of biotechnology,
Amity University Uttar Pradesh Lucknow campus for conducting this
study. This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
Conflict of Interest
The authors have no financial conflicts of interest to declare.
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