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Purification and biochemical characterization of a new thermo-stable laccase from Enterococcus faecium A2 by a three-phase partitioning method and investigation of its decolorization potential
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  • Aybüke Birge,
  • Esra Alcicek,
  • Mustafa Ozkan Baltaci,
  • Melda Şişecioğlu,
  • Ahmet Adiguzel
Aybüke Birge
Atatürk Üniversitesi
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Esra Alcicek
Atatürk Üniversitesi
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Mustafa Ozkan Baltaci
Atatürk Üniversitesi
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Melda Şişecioğlu
Atatürk Üniversitesi

Corresponding Author:[email protected]

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Ahmet Adiguzel
Atatürk Üniversitesi
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Abstract

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 enzyme activity were isolated from water samples collected from the Agri-Diyadin hotspring. The isolate with the highest laccase activity was 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 0.95-fold with 64.59% recovery using the TPP. The molecular mass of the enzyme was found by SDS-PAGE to be 50.11 kDa. Optimum pH 8.0 and optimum temperature for laccase enzyme were determined as 80 °C. It was determined that the enzyme maintained its activity at a rate of 90% after 1 h of incubation in the temperature range of 20-90 °C and showed activity and stability in a wide pH range (pH 3.0-9.0). The enzyme remained highly stable in the presence of surfactants, and increased its activity in the presence of organic solvents, Cr2+, Cu2+, and Ag+ metals. The Km and Vmax values of laccase enzyme for ABTS substrate were 0.68 mM and 5.29 μmol mL-1min-1.