Degradation of malachite green dye by Tenacibaculum sp. HMG1 isolated from Pacific deep-sea sediments
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摘要: 本文在太平洋深海沉积物中分离得到一株孔雀石绿降解菌株,鉴定命名为Tenacibaculum sp.HMG1。通过菌株生长实验和高效液相色谱的研究表明, HMG1菌株可以在20 mg/L的孔雀石绿中维持较快的生长速率,并且在12 h内可降解98.8%的孔雀石绿,这证明该菌株具有很高的孔雀石绿耐受能力和降解活性。通过基因组测序在HMG1菌株发现一条过氧化物酶基因可能参与了孔雀石绿的降解,随后利用原核表达获得了相应的重组蛋白。实验表明,该重组过氧化物酶具有极强的活性,可在1000 mg/L的孔雀石绿中发挥降解功能。本文利用液相色谱-质谱联用(LC-MS)技术对孔雀石绿的菌株降解产物和重组酶降解产物进行鉴定,并基于鉴定结果推测了两种降解途径。结果发现两种降解方式存在共同的降解途径。此外,孔雀石绿降解条件的实验结果证明重组过氧化物酶可以在低温(20℃)、复杂的pH值(6.0–9.0)、高盐度(100 mmol/L)、金属离子和EDTA等反应条件下依旧维持很高的孔雀石绿降解活性。以上实验结果表明,HMG1菌株和重组过氧化物酶均在孔雀石绿污染生物修复方面具有很大潜力。Abstract: A deep-sea bacterium from the Pacific Ocean identified as Tenacibaculum sp. HMG1 was found to have strong malachite green (MG) degradation activity. The MG tolerance and decolorizing activities of strain HMG1 were confirmed by bacterial growth and high-performance liquid chromatography (HPLC) analyses. Strain HMG1 was capable of removing 98.8% of the MG in cultures within 12 h and was able to grow vigorously at 20 mg/L MG. A peroxidase gene detected in the genome of strain HMG1 was found to be involved in the MG biodegradation process. The corresponding recombinant peroxidase (rPOD) demonstrated high degradative activity at 1 000 mg/L MG. Based on the common candidate intermediates, strain HMG1 was inferred to have one primary MG degradation pathway containing rPOD. In addition, five other candidate intermediates of the rPOD-MG degradative process were detected. The optimal conditions for MG degradation were determined and showed that strain HMG1 and the rPOD enzyme could maintain high bioactivity at a low temperature (20℃), variable pH values (6.0-9.0), higher salinities (100 mmol/L) and other factors, such as multiple metal ions, H2O2 and EDTA. MG-tolerant strain Tenacibaculum sp. HMG1 and its peroxidase have prospective applications as environmental amendments for MG degradation during coastal remediation.
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Ali L, Algaithi R, Habib H M, et al. 2013. Soybean peroxidase-mediated degradation of an azo dye—a detailed mechanistic study. BMC Biochemistry, 14:35-47 Bhunia A, Durani S, Wangikar P P.. 2001. Horseradish peroxidase catalyzed degradation of industrially important dyes. Biotechnology and Bioengineering, 72(5):562-567 Cha C J, Daniel D R, Cerniglia C E. 2001. Biotransformation of malachite green by the fungus Cunninghamella elegans. Applied and Environmental Microbiology, 67(9):4358-4360 Chen C H, Chang C F, Liu S M. 2010. Partial degradation mechanisms of malachite green and methyl violet B by Shewanella decolorationis NTOU1 under anaerobic conditions. Journal of Hazardous Materials, 177(1-3):281-289 Chen G Y, Miao S. 2010. HPLC determination and MS confirmation of malachite green, gentian violet, and their leuco metabolite residues in channel catfish muscle. Journal of Agriculture Food Chemistry, 58(12):7109-7114 Chi W J, Park D Y, Seo Y B, et al. 2014. Cloning, expression, and biochemical characterization of a novel GH16 β-agarase AgaG1 from Alteromonas sp. GNUM-1. Applied Microbiology and Biotechnology, 98(10):4545-4555 Darajeh N, Idris A, Truong P, et al. 2014. Phytoremediation potential of vetiver system technology for improving the quality of palm oil mill effluent. Advances in Materials Science and Engineering, 2014:683579 Dash H R, Mangwani N, Chakraborty J, et al. 2013. Marine bacteria:potential candidates for enhanced bioremediation. Applied Microbiology and Biotechnology, 97(2):561-571 De Souza S M A G U, Forgiarini E, de Souza A A U. 2007. Toxicity of textile dyes and their degradation by the enzyme horseradish peroxidase (HRP). Journal of Hazardous Materials, 147(3):1073-1078 Du L N, Wang S, Li G, et al. 2011. Biodegradation of malachite green by Pseudomonas sp. strain DY1 under aerobic condition:characteristics, degradation products, enzyme analysis and phytotoxicity. Ecotoxicology, 20(2):438-446 Du L N, Zhao M, Li G, et al. 2013. Biodegradation of malachite green by Micrococcus sp. strain BD15:Biodegradation pathway and enzyme analysis. International Biodeterioration & Biodegradation, 78:108-116 Forgacsa E, Cserhátia T, Oros G. 2004. Removal of synthetic dyes from wastewaters:a review. Environment International, 30(7):953-971 Garg V K, Kumar R, Gupta R. 2004. Removal of malachite green dye from aqueous solution by adsorption using agro-industry waste:a case study of Prosopis cineraria. Dyes Pigments, 62(1):1-10 Goszczynski S, Paszczynski A, Pastigrigsby M B, et al. 1994. New pathway for degradation of sulfonated azo dyes by microbial peroxidases of Phanerochaete chrysosporium and Streptomyces chromofuscus. Journal of Bacteriology, 176(5):1339-1347 Han W J, Cheng Y Y, Wang D D, et al. 2016. Biochemical characteristics and substrate degradation pattern of a novel exo-type β-agarase from the polysaccharide-degrading marine bacterium Flammeovirga sp. strain MY04. Applied and Environmental Microbiology, 82(16):4944-4954 Jadhav J P, Govindwar S P. 2006. Biotransformation of malachite green by Saccharomyces cerevisiae MTCC 463. Yeast, 23(4):315-323 Kedderis G L, Hollenberg P F. 1983. Characterization of the N-demethylation reactions catalyzed by horseradish peroxidase. Journal of Biological Chemistry, 258(13):8129-8138 Kedderis G L, Koop D R, Hollenberg P F. 1983. N-demethylation reactions catalyzed by chloroperoxidase. Journal of Biological Chemistry, 255(21):10174-10182 Kuhad R C, Kuhar S, Sharma K K, et al. 2013. Microorganisms and enzymes involved in lignin degradation vis-à-vis production of nutritionally rich animal feed:an overview. In:Kuhad R, Singh A, eds. Biotechnology for Environmental Management and Resource Recovery. India:Springer, 3-44 Littlefield N A, Blackwell B N, Hewitt C C, et al. 1985. Chronic toxicity and carcinogenicity studies of gentian violet in mice. Fundamental and Applied Toxicology, 5(5):902-912 Miwa G T, Walsh J S, Kedderis G L, et al. 1983. The use of intramolecular isotope effects to distinguish between deprotonation and hydrogen atom abstraction mechanisms in cytochrome P-450 and ceroxidase-catalyzed N-demethylation reactions. Journal of Biological Chemistry, 258(23):14445-14449 Novič M, Pihlar B, Dular M. 1988. Use of flow injection analysis based on iodometry for automation of dissolved oxygen (Winkler method) and chemical oxygen demand (dichromate method) determinations. Fresenius' Zeitschrift für analytische Chemie, 332(7):750-755 Parshetti G K, Kalme S D, Saratale G D, et al. 2006. Biodegradation of malachite green by Kocuria rosea MTCC 1532. Acta Chimica Slovenica, 53(4):492-498 Pinhassi J, Zweifel U L, Hagström A. 1997. Dominant marine bacterioplankton species found among colony-forming bacteria. Applied and Environmental Microbiology, 63(9):3359-3366 Poulos T L, Kraut J.. 1980. The stereochemistry of peroxidase catalysis. The Journal of Biological Chemistry, 255(17):8199-8205 Raghukumar C, D'Souza-Ticlo D, Verma A. 2008. Treatment of colored effluents with lignin-degrading enzymes:an emerging role of marine-derived fungi. Critical Reviews in Microbiology, 34(3-4):189-206 Ren Q, Jiang L J, Song W, et al. 2007. Isolation and identification of malachite green-degrading bacteria M3 and its degradation characteristics. Journal of Ecology and Rural Environment (in Chinese), 23(3):65-69 Robinson T, Mcmullan G, Marchant R, et al. 2001. Remediation of dyes in textile effluent:a critical review on current treatment technologies with a proposed alternative. Bioresource Technology, 77(3):247-255 Sakalle K, Rajkumar S. 2009. Isolation of crude oil degrading marine bacteria and assessment for biosurfactant production. The Internet Journal Microbiology, 7(2):1-7 Srivastava S, Sinha R, Roy D. 2004. Toxicological effects of malachite green. Aquatic Toxicology, 66(3):319-329 Tao Y B, Wang F, Meng L J, et al. 2017. Biological decolorization and degradation of malachite green by Pseudomonas sp. YB2:process optimization and biodegradation pathway. Current Microbiology, 74(10):1210-1215 Torres J M O, Cardenas C V, Moron L S, et al. 2011. Dye decolorization activities of marine-derived fungi isolated from Manila Bay and Calatagan Bay, Philippines. Philippine Journal of Science, 140(2):133-143 Von C H, Kelly S, Li Y, et al.. 2002. Species diversity improves the efficiency of mercury-reducing biofilms under changing environmental conditions. Applied and Environmental Microbiology, 68(6):2829-2837 Vu B, Chen M, Crawford R J, et al. 2009. Bacterial extracellular polysaccharides involved in biofilm formation. Molecules, 14(7):2535-2554 Wang J A, Gao F, Liu Z Z, et al. 2012. Pathway and molecular mechanisms for malachite green biodegradation in Exiguobacterium sp. MG2. PLoS One, 7(12):e51808 Weisburg W G, Barns S M, Pelletier D A, et al. 1991. 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology, 173(2):697-703 Xia Y H, Gu W C. 2000. Statistical analysis of mean sea temperatures at tidal gauge stations along Guangxi Coast. Marine Science Bulletin, 19(4):15-21 Zhang J W, Zeng R Y. 2008. Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic. Marine Biotechnology, 10(1):75-82 Zhai Y X, Zhang C, Ning J S, et al. 2007. Survey of malachite green residue in aquatic product. Marine Fisheries Research (in Chinese), 28(1):101-108 Zhang P P, Ren S Z, Xu M Y, et al. 2009. Recent advances in microbial decolorization of triphenylmethane dyes. Microbiology (in Chinese), 36(9):1410-1417
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