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Isolation, Molecular, and Metabolic Profiling of Benzene-Remediating Bacteria Inhabiting the Tannery Industry Soil

Nadia Hussain

Nadia Hussain

  • Department of Pharmaceutical Sciences, College of Pharmacy, Al Ain University, Al Ain CampusAl Ain, United Arab Emirates
  • AAU Health and Biomedical Research Center, Al Ain University, Abu Dhabi CampusAbu Dhabi, United Arab Emirates
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Hussain, Nadia
, 
Farhan Mohiuddin

Farhan Mohiuddin

School of Biochemistry and Biotechnology, University of the PunjabLahore, Pakistan
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Mohiuddin, Farhan
, 
Fatima Muccee

Fatima Muccee

School of Biochemistry and Biotechnology, University of the PunjabLahore, Pakistan
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Muccee, Fatima
, 
Saboor Muarij Bunny

Saboor Muarij Bunny

School of Biochemistry and Biotechnology, University of the PunjabLahore, Pakistan
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Bunny, Saboor Muarij
 und 
Amal H.I. Al Haddad

Amal H.I. Al Haddad

Chief Operations Office, Sheikh Shakhbout Medical City (SSMC), PureHealthAbu Dhabi, United Arab Emirates
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Al Haddad, Amal H.I.
  
26. März 2025
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Artikel-Kategorie: ORIGINAL PAPER
Online veröffentlicht: 26. März 2025
Seitenbereich: 33 - 47
Eingereicht: 20. Okt. 2024
Akzeptiert: 27. Dez. 2024
DOI: https://doi.org/10.33073/pjm-2025-003
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© 2025 Nadia Hussain et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Phylogenetic tree constructed for present study bacteria using MEGA11 software.
Phylogenetic tree constructed for present study bacteria using MEGA11 software.

Fig. 2.

Growth curve analysis of present study benzene degrading bacteria.
Growth curve analysis of present study benzene degrading bacteria.

Fig. 3.

Comparison of FTIR spectra of control and present study benzene degrading bacteria to analyze the variation of the peaks contributed by bacterial benzene degradation. A) Control, B) Paracoccus aestuarii PUB1, C) Bacillus tropicus PUB2, D) Bacillus albus PUB3, E) Bacillus subtilis PUB4, F) Bacillus cereus PUB6.
Comparison of FTIR spectra of control and present study benzene degrading bacteria to analyze the variation of the peaks contributed by bacterial benzene degradation. A) Control, B) Paracoccus aestuarii PUB1, C) Bacillus tropicus PUB2, D) Bacillus albus PUB3, E) Bacillus subtilis PUB4, F) Bacillus cereus PUB6.

Fig. 4.

Four pathways of benzene metabolism suggested in benzene-degrading bacteria in the present study based on GC-MS based identification of metabolites. A1 – benzyl alcohol dehydrogenase, B1 – benzaldehyde dehydrogenase, C1 – benzoate CoA-ligase, D1 – benzoyl-CoA 2,3-dioxygenase, A2 – methyl monooxygenase, B2 – benzoyl alcohol dehydrogenase, C2 – benzaldehyde dehydrogenase, D2 – benzoate 1,2-dioxygenase, E2 – dihydrocyclohexadiene carbohydrate dehydrogenase, A3 – benzoate dioxygenase, B3 – benzoate cis-diol dehydrogenase, A4 – benzene phenol monooxygenase
Four pathways of benzene metabolism suggested in benzene-degrading bacteria in the present study based on GC-MS based identification of metabolites. A1 – benzyl alcohol dehydrogenase, B1 – benzaldehyde dehydrogenase, C1 – benzoate CoA-ligase, D1 – benzoyl-CoA 2,3-dioxygenase, A2 – methyl monooxygenase, B2 – benzoyl alcohol dehydrogenase, C2 – benzaldehyde dehydrogenase, D2 – benzoate 1,2-dioxygenase, E2 – dihydrocyclohexadiene carbohydrate dehydrogenase, A3 – benzoate dioxygenase, B3 – benzoate cis-diol dehydrogenase, A4 – benzene phenol monooxygenase

Fig. 5.

The β-ketoadipate pathway suggested in the present study benzene degrading bacteria based on GC-MS based identification of metabolites involving this pathway. a – catechol 1,2-dioxygenase, b – muconate cycloisomerase, c-d – beta-ketoadipate:succinyl CoA transferase, e – succinyl CoA:acetyl CoA C-succinyl transferase, f – succinyl-CoA synthetase, g – succinic dehydrogenase, h – fumarase, i – malate dehydrogenase, j – citrate synthase, k – fatty acid synthase
The β-ketoadipate pathway suggested in the present study benzene degrading bacteria based on GC-MS based identification of metabolites involving this pathway. a – catechol 1,2-dioxygenase, b – muconate cycloisomerase, c-d – beta-ketoadipate:succinyl CoA transferase, e – succinyl CoA:acetyl CoA C-succinyl transferase, f – succinyl-CoA synthetase, g – succinic dehydrogenase, h – fumarase, i – malate dehydrogenase, j – citrate synthase, k – fatty acid synthase

Metabolites identified in the present study bacteria, through comparison of m/z ratios of GC-MS spectra with molecular weights of earlier reported benzene degradation metabolites_

Metabolite identified Metabolites unidentified PUB4 PUB1 Fragmentation ion m/z Molecular weight (M/w) FTlR spectral peaks (cm−1)
Benzene methylation pathway
Toluene cis-benzoate dihydrodiol, cis-benzene dihydrodiol I I 39,51,65, 77, 91,92 92.14 1452 (aromatic deformation), 2851 and 2920 (=C–H and –C–H)
Benzyl alcohol I 65, 77, 79,91, 107, 108 108 3427 (OH), 3010 (unsaturated CH), 2900 (saturated CH)
Benzaldehyde I I 29, 39,51,52, 77, 78, 105, 106 106 1637–1733 (C=O vibs)
Benzoate I I 176, 121,93, 77, 65 121 3427 (OH), 1271 (C-O-C stretching)
Catechol I I 81,66, 53, 110 110 3427 (OH), 1658 (C=C), 1250 and 1281 (C-O), 746 and 850 out of plane bending of =C–H bond of aromatic ring
Benzene degradation via benzaldehyde
Benzyl alcohol benzoate, bcnzoyl-CoA, acetyl CoA I I 65, 77, 79,91, 107,108 108 3427 (OH), 3010 (unsaturated CH), 2900 (saturated CH)
Benzaldehyde I I 29, 39,51,52, 77, 78, 105, 106 106 1637-1733 (C=O vibs)
Benzene degradation via carboxylation
Benzoate I I 176, 121,93, 77, 65 121 3427 (OH), 1271 (C–O–C stretching)
cis-1,6-dihydroxy-2,4-Cyelohexadiene-1-Carboxylie acid I I 44, 73, 149 156 2800-3300 (C–H stretch)
Catechol I I 81,66, 53 110 Same as above
Benzene degradation via phenol
Phenol I I 17, 38, 39, 40, 63, 65, 77, 93, 94 94 3427 (OH), 1650 (C=C stretch)
Catechol I 1 81,66, 53, 110 110 Same as above
β-ketoadipate pathway
Catechol 3-oxoadiphyl CoA, succinyl CoA, succinate, fumarate I I 81,66, 53, 110 110 Same as above
cis,cis-muconate I --- 77, 106, 122, 126, 138, 171, 195, 228 126 1540–1650 (asymmetric stretch of CO2-), 1360–1450 (symmetric stretch of CO2-)
3-oxoadipatc enol lactone I --- 1637–1733 (C=O vibs), 3427 (OH),
3-oxoadipate I --- 40.44, 73, 84, 87, 101, 114, 115, 160 160 1637–1733 (C=O vibs)
Palmitate I I 41,43, 57, 73, 89, 99, 117, 127, 141, 169, 183, 201,215, 229, 239, 257, 271,285,299, 313 255 2848 and 2913 (–CH3 and –CH2 vibs), 1695 (C=O), 1464 (–CH2 and –CH3), 939 (–OH out plane vibs) and 1299 (–OH in-plane vibs), 723 and 685 (–OH swinging vibs)
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