Purification and characterization of extracellular a-amylase from a thermophilic Anoxybacillus thermarum A4 strain

Baltas N. , Dincer B. , Ekinci A. P. , KOLAYLI S., ADIGÜZEL A.

BRAZILIAN ARCHIVES OF BIOLOGY AND TECHNOLOGY, vol.59, 2016 (Journal Indexed in SCI) identifier

  • Publication Type: Article / Article
  • Volume: 59
  • Publication Date: 2016
  • Doi Number: 10.1590/1678-4324-2016160346
  • Keywords: Anoxybacillus thermarum A4, Extracellular alpha-Amylase, Highly thermostable, Purification, THERMOSTABLE ALPHA-AMYLASE, BACILLUS-LICHENIFORMIS, RAW STARCH, SEQUENCE, FAMILY


alpha-Amylase from Anoxybacillus thermarum A4 was purified using ammonium sulphate precipitation and Sephadex G-100 gel filtration chromatography, with 29.8-fold purification and 74.6% yield. A4 amylase showed best performance for soluble potato starch hydrolysis at 70 degrees C and pH 5.5-10.5. A4 amylase was extremely stable at +4 degrees C, and the enzyme retained over 65% of its original alpha-amylase activity at 70 degrees C and 43% at 90 degrees C. The enzyme's Km values for soluble starch, amylopectin and amylose substrates were obtained as 0.9, 1.3 and 0.5 mg/mL, respectively. EDTA, Hg2+, B4O72-, OH-, CN-, and urea exhibited different inhibition effects; their IC50 values were identified as 8.0, 5.75, 16.5, 15.2, 8.2 and 10.9 mM, respectively. A4 amylase exhibited extreme stability toward some surfactants and perfect match for a wide variety of commercial solid and liquid detergents at 55 degrees C. So, it may be considered to be potential applications for detergent and other industrial uses.