Molecular and biochemical characterization of a recombinant endoglucanase rCKT3eng, from an extreme halophilic Haloarcula sp. strain CKT3

Safak H., Otur Ç., Kurt Kızıldoğan A.

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, cilt.151, ss.1173-1180, 2020 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 151
  • Basım Tarihi: 2020
  • Doi Numarası: 10.1016/j.ijbiomac.2019.10.161
  • Dergi Adı: INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, EMBASE, Food Science & Technology Abstracts, INSPEC, MEDLINE, Veterinary Science Database
  • Sayfa Sayıları: ss.1173-1180
  • Anahtar Kelimeler: Haloarcula sp. CKT3, Crude cellulase, rCKT3eng endoglucanase, Characterization, Specific activity, ENHANCED PRODUCTION, PROTEIN-STRUCTURE, CELLULASE, ENZYMES, EXPRESSION, ALIGNMENT, BACTERIA
  • Ondokuz Mayıs Üniversitesi Adresli: Evet

Özet

Halophilic cellulases are indispensable enzymes of heavy industrial processes as resistant biocatalysts due to high level activity at extreme conditions. In this study, crude cellulase from an extreme halophilic Haloatrula sp. CKT3 was characterized. Then, recombinant expression of putative endo-1,4-beta-glucanase gene, of CKT3 strain, in E coli BL21(DE3) was performed with the aim of obtaining highly pure, active and robust industrial enzyme for such industrial aplications. The crude cellulase had optimal activity (16.9 U/mg) at 70 degrees C, pH 7.0 and 4 M Nacl exhibiting good thermostability, high pH and halotolerance. Indeed, it is very stable in water-insoluble organic solvents with log P-o/w >= 2.13 and highly resistant to SDS (10%). Recombinant CKT3eng has a molecular weight of 36.9 kDa and 99% aminoadd identity to endo-1,4-beta-D-glucanase from / Ialoarcula at gentinensis. Its 3D structure was predicted using Phyre2 and I-TASSER. rCKT3eng enzyme provided 31.6 U/mg activity at optimal 50 degrees C. pH 7.0 and 3 M NaCl. In addition to its quite similar stability values and resistance to organic solvents and SDS, rCKT3eng has superiority over crude enzyme with 1.87-fold higher specific activity. Therefore, rCKT3eng offers a promising enzyme for industrial use with its valuable activity and stability in extreme conditions. (C) 2019 Elsevier B.V. All rights reserved.