Cloning, expression and biochemical characterization of lignin-degrading DyP-type peroxidase from Bacillus sp. Strain BL5

Khan S. I., Zada N. S., Sahinkaya M., Colak D. N., Ahmed S., Hasan F., ...Daha Fazla

ENZYME AND MICROBIAL TECHNOLOGY, cilt.151, 2021 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 151
  • Basım Tarihi: 2021
  • Doi Numarası: 10.1016/j.enzmictec.2021.109917
  • Dergi Adı: ENZYME AND MICROBIAL TECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Artic & Antarctic Regions, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Chimica, Compendex, EMBASE, Environment Index, Food Science & Technology Abstracts, INSPEC, MEDLINE, Veterinary Science Database
  • Anahtar Kelimeler: DyP-type peroxidase, Bacillus sp., Black liquor, Lignin degradation, FT-IR, SEM, GC-MS, KRAFT LIGNIN, DECOLORIZING PEROXIDASE, THERMOBIFIDA FUSCA, HYDROGEN-PEROXIDE, POLYMERIC LIGNIN, DEGRADATION, IDENTIFICATION, MECHANISM, VALORIZATION, PYROLYSIS
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

Lignin is a major byproduct of pulp and paper industries, which is resistant to depolymerization due to its heterogeneous structure. The enzymes peroxidases can be utilized as potent bio-catalysts to degrade lignin. In the current study, an Efeb gene of 1251bp encoding DyP-type peroxidase from Bacillus sp. strain BL5 (DyPBL5) was amplified, cloned into a pET-28a (+) vector and expressed in Escherichia coli BL21 (DE3) cells. A 46 kDa protein of DyPBL5 was purified through ion-exchange chromatography. Purified DyPBL5 was active at wide temperature (25-50 degrees C) and pH (3.0-8.0) range with optimum activity at 35 degrees C and pH 5.0. Effects of different chemicals on DyPBL5 were determined. The enzyme activity was strongly inhibited by SDS, DDT and beta-mercaptoethanol, whereas stimulated in the presence of organic solvents such as methanol and ethanol. The kinetic parameters were determined and K-m, V-max and K-cat values were 1.06 mM, 519.75 mu mol/min/mg and 395 S-1, respectively. Docking of DyPBL5 with ABTS revealed that, Asn 244, Arg 339, Asp 383 and Thr 389 are putative amino acids, taking part in the oxidation of ABTS. The recombinant DyPBL5 resulted in the reduction of lignin contents up to 26.04 %. The SEM and FT-IR analysis of test samples gave some indications about degradation of lignin by DyPBL5. Various low molecular weight lignin degradation products were detected by analyzing the samples through gas chromatography mass spectrometry. High catalytic efficiency and lignin degradation rate make DyPBL5 an ideal bio-catalyst for remediation of lignin-contaminated sites.