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
prolekare.web.journal.doi_sk: 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


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