BRAZILIAN ARCHIVES OF BIOLOGY AND TECHNOLOGY A N I N T E R N A T I O N A L J O U R N A L Purification and characterization of extracellular α-amylase from a thermophilic Anoxybacillus thermarum A4 strain

ABSTRAC

Characterization and inhibition studies of carbonic anhydrase from gill of Russian Sturgeon Fish (Acipenser gueldenstaedtii)

KURTOGLU, ILKER ZEKI/0000-0002-4214-7997WOS: 000385270300104PubMed: 26290152An alpha-carbonic anhydrase (CA, EC 4.2.1.1) was purified and characterized kinetically from gill of Acipenser gueldenstaedtii as an endangered sturgeon species. the carbonic anhydrase was purified 66-folds with yield 20.7% by Sepharose-4B-L-tyrosine-sulfanilamide affinity column and the specific activity was determined as 222.2 EU/mg protein. K-m and V-max kinetic values for gill carbonic anhydrase were calculated by a Lineweaver-Burk graph using p-nitrophenol acetate (p-NPA) as a substrate, and was defined as 2.5mM and 5 x 10(6) mu M/min, respectively. It was observed that CA from the sturgeon gill in the presence of the sulfanilamide and acetazolamide as an inhibitor had very low IC50 values such as 13.0 and 0.1 mu M, respectively. in addition, it was determined that the enzyme was inhibited by Fe-2+,Fe- Co-2+,Co- Ni2+, and Zn2+-Ba2+ with the IC50 values of 0.2, 1.7, 1.2, and 1.1 mM, respectively.Recep Tayyip Erdogan UniversityRecep Tayyip Erdogan UniversityThis work was financially supported by the Recep Tayyip Erdogan University

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

Adiguzel, Ahmet/0000-0001-8848-6647WOS: 000396245100008alpha-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

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

ABSTRACT α-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 °C and pH 5.5-10.5. A4 amylase was extremely stable at +4 °C, and the enzyme retained over 65% of its original α-amylase activity at 70 °C and 43% at 90 °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+, B4O7 2-, 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 °C. So, it may be considered to be potential applications for detergent and other industrial uses