Production, purification and evaluation of biodegradation potential of PHB depolymerase of Stenotrophomonas sp. RZS7
Autoři:
R. Z. Sayyed aff001; S. J. Wani aff001; Abdullah A. Alarfaj aff002; Asad Syed aff002; Hesham Ali El-Enshasy aff003
Působiště autorů:
Department of Microbiology, PSGVP Mandal’s, Arts, Science, and Commerce College, SHAHADA, Maharashtra, India
aff001; Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia
aff002; Institute of Bioproducts Development (IBD), Universiti Teknologi Malaysia (UTM), Johor Bahru, Johor, Malaysia
aff003; City of Scientific Research and Technology Applications, New Burg Al Arab, Alexandria, Egypt
aff004
Vyšlo v časopise:
PLoS ONE 15(1)
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pone.0220095
Souhrn
There are numerous reports on poly-β-hydroxybutyrate (PHB) depolymerases produced by various microorganisms isolated from various habitats, however, reports on PHB depolymerase production by an isolate from plastic rich sites scares. Although PHB has attracted commercial significance, the inefficient production and recovery methods, inefficient purification of PHB depolymerase and lack of ample knowledge on PHB degradation by PHB depolymerase have hampered its large scale commercialization. Therefore, to ensure the biodegradability of biopolymers, it becomes imperative to study the purification of the biodegrading enzyme system. We report the production, purification, and characterization of extracellular PHB depolymerase from Stenotrophomonas sp. RZS7 isolated from a dumping yard rich in plastic waste. The isolate produced extracellular PHB depolymerase in the mineral salt medium (MSM) at 30°C during 4 days of incubation under shaking. The enzyme was purified by three methods namely ammonium salt precipitation, column chromatography, and solvent purification. Among these purification methods, the enzyme was best purified by column chromatography on the Octyl-Sepharose CL-4B column giving optimum yield (0.7993 Umg-1mL-1). The molecular weight of purified PHB depolymerase was 40 kDa. Studies on the assessment of biodegradation of PHB in liquid culture medium and under natural soil conditions confirmed PHB biodegradation potential of Stenotrophomonas sp. RZS7. The results obtained in Fourier-Transform Infrared (FTIR) analysis, High-Performance Liquid Chromatography (HPLC) study and Gas Chromatography Mass-Spectrometry (GC-MS) analysis confirmed the biodegradation of PHB in liquid medium by Stenotrophomonas sp. RZS7. Changes in surface morphology of PHB film in soil burial as observed in Field Emission Scanning Electron Microscopy (FESEM) analysis confirmed the biodegradation of PHB under natural soil environment. The isolate was capable of degrading PHB and it resulted in 87.74% biodegradation. A higher rate of degradation under the natural soil condition is the result of the activity of soil microbes that complemented the biodegradation of PHB by Stenotrophomonas sp. RZS7.
Klíčová slova:
Esters – Gas chromatography-mass spectrometry – High performance liquid chromatography – Mass spectra – Polymers – Scanning electron microscopy – Biodegradation – Ammonium sulfate precipitation
Zdroje
1. Sayyed RZ, Gangurde NS, Chincholkar SB, Hypochlorite digestion method for efficient recovery of PHB from A. feacalis, Indian J Microbiol, 2009; 49:230–232. doi: 10.1007/s12088-009-0036-7 23100774
2. Gangurde NS, Patil YP, Jain R, Sayyed RZ, Poly-β-hydroxybutyrate biodegradation by mixed culture population vis-à-vis single culture population under varying environmental conditions: a new approach, Indian J Exp Biol, 2017; 55:311–320.
3. Wani SJ, Shaikh SS, Tabassum B, Thakur A, Gulati A, Stenotrophomonas sp. RZS7, a novel PHB degrader isolated from plastic contaminated soil in Shahada, Maharashtra, Western India, 3 Biotech, 2016; 6:179.
4. Wani SJ, Sayyed RZ, Production, efficient recovery and partial characterization of biodegradable polymer produced by soil Streptomyces, Indian J Biotechnol, 2016; 15: 127–129.
5. Gangurde NS, Sayyed RZ, Shashi K, Gulati A, Development of eco-friendly bioplastic like PHB from distillery effluent microorganisms, Env Sci Poll Res, 2013; 20:488–497.
6. Soam A, Singh A, Singh R, and Shahi S, Optimization of culture conditions for biopolymer producing Bacillus mycoides (wss2) bacteria from sewage. Curr Discov 2012; 1:27–32.
7. Kumar MS, Mudliar SN, Reddy KMK, Chakraborti T, Production of Biodegradable Plastic from Activated Sludge Generated from the Food Processing Industrial Wastewater Treatment Plant, Biores Technol, 2004; 95:327–330.
8. Sudesh K, Abe H, Doi Y, Synthesis and properties of polyhydroxyalkanoates: biological polyesters. Progress in Polym Sci, 2000; 25:1503–1555.
9. Jendrossek D and Handrick R, Microbial degradation of polyhydroxyalkanoates. Ann. Rev Microbiol 2002; 56:403–432.
10. Iyer S, Shah R, Sharma A, Jendrossek D, Desai A, Purification of Aspergillus fumigatus (Pdf1) poly(β-hydroxybutyrate) (PHB) depolymerase using a new, single-step substrate affinity chromatography method: characterization of the PHB depolymerase exhibiting novel self-aggregation behavior. J Polym Environ 2000; 8:197–203.
11. Sadocco PS, Nocerino E, Dubini-Paglia A, Seres EG, Characterization of a poly(3-hydroxybutyrate) depolymerase from Aureobacterium saperdae: Active site and kinetics of hydrolysis studies. J Environ Polym Degrad 1997; 5:57–65.
12. Papaneophytou CP, Pantazaki AA, Kyriakidis DA, An extracellular polyhydroxybutyrate depolymerase in thermus thermophilus hb8. Appl Microbiol Biotechnol, 2009; 83:659–668. doi: 10.1007/s00253-008-1842-2 19214501
13. Hsu KJ, Tseng M, Don TM, Yang MK, Biodegradation of Poly(β-hydroxybutyrate) by a novel isolate of Streptomyces bangladeshensis 77T-4. Bot Stud, 2012; 53:307–313.
14. Han JS, Kim MN, Purification and characterization of extracellular Poly (3-hydroxybutyrate) Depolymerase from Penicillium simplicissimum LAR13. The J Microbiol, 2002; 40:20–25.
15. Kobayashi T, Sugiyama A, Kawase Y, Saito T, Mergaert J, Swings J, Biochemical and genetic characterization of an extracellular Poly(3-Hydroxybutyrate) depolymerase from Acidovorax sp. strain TP4. J Env Pol Deg 1999; 7:9–18.
16. Kim HJ, Nam JS, Bae KS, Rhee YH, Characterization of an extracellular medium-chain length poly (3-hydroxyalkanoate) depolymerase from Streptomyces sp. KJ-72, Antony van Leeuwenhoek 2003; 83:183–189.
17. Lucas N, Bieniame C, Belloy C, Queneudec M, Silvestre F, Nava-Saucedo JE, Polymer biodegradation: mechanisms and estimation techniques. Chemosphere 2008; 73:429–442. doi: 10.1016/j.chemosphere.2008.06.064 18723204
18. Sudesh K, Abe H, Doi Y, Synthesis and properties of polyhydroxyalkanoates: biological polyesters. Prog Polym Sci, 2000; 25:1503–1555.
19. Wani SJ, Shaikh SS, Sayyed RZ, Medium optimization for PHB depolymerase production by Stenotrophomonas maltophilia using Plackett Burman design & response surface methodology. Intntl J of Scientific & Engg Res, 2015; 6:818–829.
20. Lowry OH, Rosebrough NJ, Farr AL and Randall RJ, Protein measurement with the Folin phenol reagent. J Biol Chem; 1951, 193:265–275. 14907713
21. Liang W, Wenfu Z, Xiaojuan W, Xianyu C, Guo QC, Kaitian X, Processability Modifications of Poly(3-hydroxybutyrate) by Plasticizing, Blending, and Stabilizing. J Appl Poly Sci, 2008; 107:166–173.
22. Law JH, Slepecky W, Assay of poly-β-hydroxbutyric acid. J Bacteriol, 1960; 82:33–36.
23. Dalev PG, Patil RD, Mark JE, Vassileva E, Fakirao S, Biodegradation of chemically modified gelatin film in soil. J Applied Poly Sci, 2000; 78:1341–1347.
24. Mustafa K, Helen BJ, Don M, Quantitative determination of the biodegradable polymer Poly β hydroxybutyrate in a recombinant E. coli strain by use of Mid infrared spectroscopy and multivariative statistics. Appl and Env Microbiol, 2000; 66(8): 3415–3420.
25. Gebauer B, Jendrossek D, Assay of poly(3-hydroxybutyrate) depolymerase activity and product determination. Appl Environ Microbiol, 2006; 72: 6094–6100. doi: 10.1128/AEM.01184-06 16957234
26. Nehal T, Ujjval T, Patel KC, Biosynthesis of medium chain length poly(3-hydroxyalkanoates) (mcl-PHAs) by Comamonas testosterone during cultivation on vegetable oils. Biores Technol, 2005; 96:1843–1850.
27. Parker RE, Continuous distribution: tests of significance. In: Parker RE(ed) Introductory statistics for biology, 2nd Edn; 1979; Cambridge University Press, London, pp 18–42.
28. Zhou H, Wang Z, Chen S, Liu D, Xia H, Purification and characterization of extracellular poly(β-hydroxybutyrate) depolymerase from Penicillium sp. DS9701-D2. Polymer-Plastics Technol Engg, 2008; 48: 58–63.
29. Shivakumar S, Jagadish SJ, Zatakia H, Dutta J, Purification, characterization and kinetic studies of a novel poly (β) hydroxybutyrate (PHB) depolymerase PhaZ Pen from Penicillium citrinum S2. Appl Biochem and Biotechnol, 2011; 164: 1225–1236.
30. Shah AA, Hasan F, Hameed A, Ahmed S, Isolation and characterization of poly (3-hydroxybutyrate-co-3-hydroxy valerate) degrading actinomycetes and purification of PHBV depolymerase from newly isolated Streptoverticillium kashmirense AF1. Annals of Microbiol 2007;57: 583–588
31. Kim DY, Yun JH, Kim HW, Bae KS, Rhee YH, Purification and characterization of poly (3-hydroxybutyrate) depolymerase from a fungal isolate, Emericellopsis minima W2. J Microbiol 2002; 40: 129–133.
32. Varda M, Nishith D, Darshan M, Production and evaluation of microbial plastic for its degradation capabilities. J Environ Res Develop, 2014; 8(4): 934–940.
33. McLafferty FW, Turecek F, Interpretation of mass spectra Fourth edition. University Science Books, Mill Valley, California, 1993; pp 211–216.
34. Wennan H, Weidong T, Guang Z, Guo QC, Zengming Z, Production of novel polyhydroxyalkanoates by Pseudomonas stutzeri 1317 from glucose and soybean oil. FEMS Microbiol Lett, 1998; 169: 45–49.
35. Gautam N, Kaur I, Soil burial biodegradation studies of starch grafted polyethylene and identification of Rhizobium meliloti therefrom. J Env Chem and Ecotoxicol, 2013; 5(6):147–158.
36. Calabia BP, Tokiwa Y, A novel PHB depolymerase from a thermophilic Streptomyces sp. Biotechnol Lett, 2006; 28:383–388. doi: 10.1007/s10529-005-6063-5 16614903
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