Cloning, biochemical characterization and molecular docking of novel thermostable beta-glucosidase BglA9 from Anoxybacillus ayderensis A9 and its application in de-glycosylation of Polydatin

Zada N. S., Belduz A. O., Güler H. İ., Sahinkaya M., Khan S. I., Saba M., ...Daha Fazla

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, cilt.193, ss.1898-1909, 2021 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 193
  • Basım Tarihi: 2021
  • Doi Numarası: 10.1016/j.ijbiomac.2021.11.021
  • 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.1898-1909
  • Anahtar Kelimeler: Polydatin, Enzymatic de-glycosylation, Anoxybacillus ayderensis A9, POLYGONUM-CUSPIDATUM, HYDROLYSIS, GLYCOSIDASE, PURIFICATION, RESVERATROL, BIOTRANSFORMATION, EXPRESSION, TOLERANCE, PICEID, THERMOPHILUM
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

This study reports a novel BglA9 gene of 1345 bp encoding beta-glucosidase from Anoxybacillus ayderensis A9, which was amplified and expressed in E. coli BL21 (DE3): pLysS cells, purified with Ni-NTA column having molecular weight of 52.6 kDa and was used in the bioconversion of polydatin to resveratrol. The kinetic parameters values using pNPG as substrate were Km (0.28 mM), Vmax (43.8 mu mol/min/mg), kcat (38.43 s-1) and kcat/Km (135.5 s-1 mM-1). The BglA9 was active in a broad pH range and had an activity half-life around 24 h at 50 degrees C. The deglycosylation efficiency of BglA9 for polydatin was determined by estimating the amount of glucose released after enzymatic reaction by a dinitrosalicylic acid (DNS) assay. The kinetic parameters of BglA9 for polydatin were 5.5 mM, 20.84 mu mol/min/mg, 18.28 s- 1and 3.27 s-1 mM-1 for Km, Vmax, kcat, and kcat/Km values, respectively. The Ki value for glucose was determined to be 1.7 M. The residues Gln19, His120, Glu355, Glu409, Glu178, Asn222 may play a crucial role in the deglycosylation as revealed by the 3D structure of enzyme docked with polydatin.