Abstract
Endophytic microorganisms can metabolize organic contaminants and assist in plant growth, thus facilitating the phytoremediation of polluted environments. An endophytic bacterium capable of decoloring malachite green (MG) was isolated from the leaves of the wetland plant Suaeda salsa and was identified as Klebsiella aerogenes S27. Complete decolorization of MG (100 mg/l) was achieved in 8 h at 30 °C and pH 7.0. Ultraviolet-visible spectroscopy and Fourier-transform infrared spectroscopy analyses indicated the degradation of MG by the isolate. The enzymic assays of the strain showed the triphenylmethane reductase (TMR) activity. A gene encoding putative TMR-like protein (named as KaTMR) was cloned and heterologously expressed in Escherichia coli. KaTMR showed only 42.6–43.3% identities in amino acids compared with well-studied TMRs, and it phylogenetically formed a new branch in the family of TMRs. The degraded metabolites by recombinant KaTMR were detected by liquid chromatography-mass spectrometry, showing differences from the products of reported TMRs. The biotransformation pathway of MG was proposed. Phytotoxicity studies revealed the less-toxic nature of the degraded metabolites compared to the dye. This study presented the first report of an endophyte on the degradation and detoxification of triphenylmethane dye via a novel oxidoreductase, thus facilitating the study of the plant-endophyte symbiosis in the bioremediation processes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahemad M (2015) Phosphate-solubilizing bacteria-assisted phytoremediation of metalliferous soils: a review. 3 Biotech 5:111–121. https://doi.org/10.1007/s13205-014-0206-0
An SY, Min SK, Cha IH, Choi YL, Cho YS, Kim CH, Lee YC (2002) Decolorization of triphenylmethane and azo dyes by Citrobacter sp. Biotechnol Lett 24:1037–1040. https://doi.org/10.1023/a:1015610018103
Ayed L, Chaieb K, Cheref A, Bakhrouf A (2008) Biodegradation of triphenylmethane dye malachite green by Sphingomonas paucimobilis. World J Microbiol Biotechnol 25:705–711. https://doi.org/10.1007/s11274-008-9941-x
Ayed L, Chaieb K, Cheref A, Bakhrouf A (2010) Biodegradation and decolorization of triphenylmethane dyes by Staphylococcus epidermidis. Desalination 260:137–146. https://doi.org/10.1016/j.desal.2010.04.052
Azmi W, Sani RK, Banerjee UC (1998) Biodegradation of triphenylmethane dyes. Enzym Microb Technol 22:185–191
Bañuelos JA, García-Rodríguez O, El-Ghenymy A, Rodríguez-Valadez FJ, Godínez LA, Brillas E (2016) Advanced oxidation treatment of malachite green dye using a low cost carbon-felt air-diffusion cathode. J Environ Chem Eng 4:2066–2075. https://doi.org/10.1016/j.jece.2016.03.012
Casas N, Parella T, Vicent T, Caminal G, Sarra M (2009) Metabolites from the biodegradation of triphenylmethane dyes by Trametes versicolor or laccase. Chemosphere 75:1344–1349. https://doi.org/10.1016/j.chemosphere.2009.02.029
Culp SJ, Beland FA (1996) Malachite green: a toxicological review. J Am Coll Toxicol 15:219–238
Daneshvar N, Ayazloo M, Khataee AR, Pourhassan M (2007) Biological decolorization of dye solution containing malachite green by microalgae Cosmarium sp. Bioresour Technol 98:1176–1182. https://doi.org/10.1016/j.biortech.2006.05.025
Du LN, Li G, Zhao YH, Xu HK, Wang Y, Zhou Y, Wang L (2015) Efficient metabolism of the azo dye methyl orange by Aeromonas sp. strain DH-6: characteristics and partial mechanism. Int Biodeterior Biodegrad 105:66–72. https://doi.org/10.1016/j.ibiod.2015.08.019
Du LN, Zhao M, Li G, Zhao XP, Zhao YH (2012) Highly efficient decolorization of malachite green by a novel Micrococcus sp. strain BD15. Environ Sci Pollut Res Int 19:2898–2907. https://doi.org/10.1007/s11356-012-0796-1
Dutta V, Elhanafi D, Osborne J, Martinez MR, Kathariou S (2014) Genetic characterization of plasmid-associated triphenylmethane reductase in Listeria monocytogenes. Appl Environ Microbiol 80:5379–5385. https://doi.org/10.1128/aem.01398-14
Eapen S, Singh S, D'Souza SF (2007) Advances in development of transgenic plants for remediation of xenobiotic pollutants. Biotechnol Adv 25:442–451. https://doi.org/10.1016/j.biotechadv.2007.05.001
Finn RD, Coggill P, Eberhardt RY, Eddy SR, Mistry J, Mitchell AL, Potter SC, Punta M, Qureshi M, Sangrador-Vegas A, Salazar GA, Tate J, Bateman A (2016) The Pfam protein families database: towards a more sustainable future. Nucleic Acids Res 44:D279–D285. https://doi.org/10.1093/nar/gkv1344
Fu XY, Zhao W, Xiong AS, Tian YS, Zhu B, Peng RH, Yao QH (2013) Phytoremediation of triphenylmethane dyes by overexpressing a Citrobacter sp. triphenylmethane reductase in transgenic Arabidopsis. Appl Microbiol Biotechnol 97:1799–1806. https://doi.org/10.1007/s00253-012-4106-0
Gao F, Ding H, Shao L, Xu X, Zhao Y (2015) Molecular characterization of a novel thermal stable reductase capable of decoloration of both azo and triphenylmethane dyes. Appl Microbiol Biotechnol 99:255–267. https://doi.org/10.1007/s00253-014-5896-z
Huan M, Lian-Tai L, Cai-Fang Y, Jin-Jin S, Yuan G, Hong Q, Shun-Peng L (2011) Biodegradation of malachite green by strain Pseudomonas sp. K9 and cloning of the tmr2 gene associated with an ISPpu12. World J Microbiol Biotechnol 27:1323–1329. https://doi.org/10.1007/s11274-010-0580-7
Iyer R, Iken B, Damania A (2017) Whole genome of Klebsiella aerogenes PX01 isolated from San Jacinto River sediment west of Baytown, Texas reveals the presence of multiple antibiotic resistance determinants and mobile genetic elements. Genomics Data 14:7–9. https://doi.org/10.1016/j.gdata.2017.07.012
Jang MS, Lee YM, Kim CH, Lee JH, Kang DW, Kim SJ, Lee YC (2005) Triphenylmethane reductase from Citrobacter sp. strain KCTC 18061P: purification, characterization, gene cloning, and overexpression of a functional protein in Escherichia coli. Appl Environ Microbiol 71:7955–7960. https://doi.org/10.1128/aem.71.12.7955-7960.2005
Kagalkar AN, Jadhav MU, Bapat VA, Govindwar SP (2011) Phytodegradation of the triphenylmethane dye malachite green mediated by cell suspension cultures of Blumea malcolmii Hook. Bioresour Technol 102:0312e10318–0312e10318. https://doi.org/10.1016/j.biortech.2011.08.101
Khan R, Bhawana P, Fulekar MH (2013) Microbial decolorization and degradation of synthetic dyes: a review. Rev Environ Sci Biotechnol 12:75–97. https://doi.org/10.1007/s11157-012-9287-6
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948. https://doi.org/10.1093/bioinformatics/btm404
Li L, Peng AH, Lin ZZ, Zhong HP, Chen XM, Huang ZY (2017) Biomimetic ELISA detection of malachite green based on molecularly imprinted polymer film. Food Chem 229:403–408. https://doi.org/10.1016/j.foodchem.2017.02.090
Li LT, Hong Q, Yan X, Fang GH, Ali SW, Li SP (2009) Isolation of a malachite green-degrading Pseudomonas sp. MDB-1 strain and cloning of the tmr2 gene. Biodegradation 20:769–776. https://doi.org/10.1007/s10532-009-9265-z
Liu Q, Liu R, Ma Y, Song J (2018) Physiological and molecular evidence for Na+ and Cl− exclusion in the roots of two Suaeda salsa populations. Aquat Bot 146:1–7. https://doi.org/10.1016/j.aquabot.2018.01.001
Liu W, Hou J, Wang Q, Yang H, Luo Y, Christie P (2015) Collection and analysis of root exudates of Festuca arundinacea L. and their role in facilitating the phytoremediation of petroleum-contaminated soil. Plant Soil 389:109–119. https://doi.org/10.1007/s11104-014-2345-9
Lorenzo M, Moldes D, Rodriguez Couto S, Sanroman MA (2005) Inhibition of laccase activity from Trametes versicolor by heavy metals and organic compounds. Chemosphere 60:1124–1128. https://doi.org/10.1016/j.chemosphere.2004.12.051
Matpang P, Sriuttha M, Piwpuan N (2017) Effects of malachite green on growth and tissue accumulation in pak choy (Brassica chinensis Tsen & Lee). Agr Nat Resour 51:96–102
Meagher RB (2000) Phytoremediation of toxic elemental and organic pollutants. Curr Opin Plant Biol 3:153–162. https://doi.org/10.1016/S1369-5266(99)00054-0
Mosa KA, Saadoun I, Kumar K, Helmy M, Dhankher OP (2016) Potential biotechnological strategies for the cleanup of heavy metals and metalloids. Front Plant Sci 7:303. https://doi.org/10.3389/fpls.2016.00303
Mukherjee T, Das M (2014) Degradation of malachite green by Enterobacter asburiae strain XJUHX-4TM. Clean-Soil Air Water 42:849–856. https://doi.org/10.1002/clen.201200246
Novotny C, Rawal B, Bhatt M, Patel M, Sasek V, Molitoris HP (2001) Capacity of Irpex lacteus and Pleurotus ostreatus for decolorization of chemically different dyes. J Biotechnol 89:113–122
Parshetti GK, Kalme SD, Saratale GD, Govindwar SP (2006) Biodegradation of malachite green by Kocuria rosea MTCC 1532. Acta Chim Slov 53:492–498
Persson B, Kallberg Y, Oppermann U, Jornvall H (2003) Coenzyme-based functional assignments of short-chain dehydrogenases/reductases (SDRs). Chem Biol Interact 143-144:271–278
Rai MS, Rama Bhat P, Prajna PS, Jayadev K, Venkatakrishna Rao PS (2014) Degradation of malachite green and congo red using Aloe barabadensis mill. Extract Int J Curr Microbiol App Sc 3:330e340
Ren S, Guo J, Zeng G, Sun G (2006) Decolorization of triphenylmethane, azo, and anthraquinone dyes by a newly isolated Aeromonas hydrophila strain. Appl Microbiol Biotechnol 72:1316–1321. https://doi.org/10.1007/s00253-006-0418-2
Rosenblueth M, Martinez L, Silva J, Martinez-Romero E (2004) Klebsiella variicola, a novel species with clinical and plant-associated isolates. Syst Appl Microbiol 27:27–35. https://doi.org/10.1078/0723-2020-00261
Rudd T, Sterritt RM, Lester JN (1983) Mass balance of heavy metal uptake by encapsulated cultures of Klebsiella aerogenes. Microb Ecol 9:261–272. https://doi.org/10.1007/BF02097741
Schlüter A, Krahn I, Kollin F, Bönemann G, Stiens M, Szczepanowski R, Schneiker S, Puhler A (2007) IncP-1-β plasmid pGNB1 isolated from a bacterial community from a wastewater treatment plant mediates decolorization of triphenylmethane dyes. Appl Environ Microbiol 73:6345–6350. https://doi.org/10.1128/aem.01177-07
Shedbalkar U, Dhanve R, Jadhav J (2008) Biodegradation of triphenylmethane dye cotton blue by Penicillium ochrochloron MTCC 517. J Hazard Mater 157:472–479. https://doi.org/10.1016/j.jhazmat.2008.01.023
Shin SH, Kim S, Kim JY, Lee S, Um Y, Oh MK, Kim YR, Lee J, Yang KS (2012) Complete genome sequence of Enterobacter aerogenes KCTC 2190. J Bacteriol 194:2373–2374. https://doi.org/10.1128/JB.00028-12
Song J, Wang B (2015) Using euhalophytes to understand salt tolerance and to develop saline agriculture: Suaeda salsa as a promising model. Ann Bot.115:541-553. https://doi.org/10.1093/aob/mcu194
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Wang J, Gao F, Liu Z, Qiao M, Niu X, Zhang KQ, Huang X (2012) Pathway and molecular mechanisms for malachite green biodegradation in Exiguobacterium sp. MG2. PLoS One 7:e51808. https://doi.org/10.1371/journal.pone.0051808
Wei CY, Lin L, Luo LJ, Xing YX, Hu CJ, Yang LT, Li YR, An Q (2014) Endophytic nitrogen-fixing Klebsiella variicola strain DX120E promotes sugarcane growth. Biol Fertil Soils 50:657–666. https://doi.org/10.1007/s00374-013-0878-3
Wu HF, Liu XL, Zhao JM, Yu JB (2012) Toxicological responses in halophyte Suaeda salsa to mercury under environmentally relevant salinity. Ecotox Environ Safe 85:64–71. https://doi.org/10.1016/j.ecoenv.2012.03.016
Wu L, Lin ZZ, Zhong HP, Chen XM, Huang ZY (2017) Rapid determination of malachite green in water and fish using a fluorescent probe based on CdTe quantum dots coated with molecularly imprinted polymer. Sensors Actuators B Chem 239:69–75. https://doi.org/10.1016/j.snb.2016.07.166
Wu Y, Xiao X, Xu C, Cao D, Du D (2013) Decolorization and detoxification of a sulfonated triphenylmethane dye aniline blue by Shewanella oneidensis MR-1 under anaerobic conditions. Appl Microbiol Biotechnol 97:439–7446. https://doi.org/10.1007/s00253-012-4476-3
Yang X, Zheng J, Lu Y, Jia R (2016) Degradation and detoxification of the triphenylmethane dye malachite green catalyzed by crude manganese peroxidase from Irpex lacteus F17. Environ Sci Pollut Res Int 23:9585–9597. https://doi.org/10.1007/s11356-016-6164-9
Zeng J, Eckenrode Heather M, Dounce Susan M, Dai HL (2013) Time-resolved molecular transport across living cell membranes. Biophys J 104:139–145. https://doi.org/10.1016/j.bpj.2012.11.3814
Zhou Q (2001) Chemical pollution and transport of organic dyes in water-soil-crop systems of the Chinese Coast. Bull Environ Contam Toxicol 66:784–793. https://doi.org/10.1007/s00128-001-0077-z
Zhou X, Zhang J, Pan Z, Li D (2018) Review of methods for the detection and determination of malachite green and leuco-malachite green in aquaculture. Crit Rev Anal Chem 14:1–20. https://doi.org/10.1080/10408347.2018.1456314
Funding
This study was funded by National Natural Science Foundation of China (Grant No. 31640002), the Natural Science Foundation of Shandong Province (Grant No. ZR2015JL013), the China Postdoctoral Science Foundation (Grant No. 2016 M600551), and the International Postdoctoral Exchange Fellowship, China (Grant No. 20170058).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Research involving human participants and/or animals
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(PDF 399 kb)
Rights and permissions
About this article
Cite this article
Shang, N., Ding, M., Dai, M. et al. Biodegradation of malachite green by an endophytic bacterium Klebsiella aerogenes S27 involving a novel oxidoreductase. Appl Microbiol Biotechnol 103, 2141–2153 (2019). https://doi.org/10.1007/s00253-018-09583-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-018-09583-0