Purification and biochemical characterization of a new thermostable laccase from Enterococcus faecium A2 by a three-phase partitioning method and investigation of its decolorization potential

Birge A., Alcicek E. A., Baltacı M. Ö., Şişecioğlu M., Adıgüzel A.

ARCHIVES OF MICROBIOLOGY, cilt.204, sa.8, 2022 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 204 Sayı: 8
  • Basım Tarihi: 2022
  • Doi Numarası: 10.1007/s00203-022-03054-x
  • Dergi Adı: ARCHIVES OF MICROBIOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, CAB Abstracts, Chemical Abstracts Core, EMBASE, Environment Index, Food Science & Technology Abstracts, MEDLINE, Veterinary Science Database
  • Anahtar Kelimeler: Enterococcus faecium, Laccase, Three-phase partitioning, Biochemical characterization, Decolorization, PH-STABLE LACCASE, MULTICOPPER OXIDASE, TRAMETES-VERSICOLOR, ENZYMATIC DECOLORIZATION, STREPTOMYCES-COELICOLOR, PYCNOPORUS-SANGUINEUS, TOLERANT LACCASE, ALKALINE LACCASE, TEXTILE DYES, EXPRESSION
  • Atatürk Üniversitesi Adresli: Evet

Özet

Three-phase partitioning (TPP) is a simple, fast, cost-effective, and highly efficient process that can be used in the purification of laccases. In this study, microorganisms with laccase activity were isolated from water samples collected from the Agri-Diyadin hot spring. The isolate with the highest laccase activity was found to be the A2 strain. As a result of molecular (16S rRNA sequence) and conventional (morphological, biochemical, and physiological) analyses, it was determined that the A2 isolate was 99% similar to Enterococcus faecium (Genbank number: MH424896). The laccase was purified to 4.9-fold with 110% recovery using the TPP. The molecular mass of the enzyme was found by SDS-PAGE to be 50.11 kDa. Optimum pH 6.0 and optimum temperature for laccase were determined as 80 degrees C. The laccase exhibited pH stability over a wide range (pH 3.0-9.0) and a high thermostability, retaining over 90% of its activity after 1 h of incubation at 20-90 degrees C. The laccase exhibited high thermostability, with a heat inactivation half-life of approximately 24 h at 80 degrees C. The enzyme remained highly stable in the presence of surfactants and increased its activity in the presence of organic solvents, Cr2+, Cu2+, and Ag+ metal ions. The K-m, V-max, k(cat), and k(cat)/K-m values of laccase for 2,2 '-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) substrate were 0.68 mM, 5.29 mu mol mL(-1) min(-1), 110.2 s(-1), and 162.1 s(-1) mM(-1), respectively.