Biochemical characterization of an acid phosphatase from Thermus thermophilus

• Published in: Bioscience, biotechnology, and biochemistry Vol. 74; no. 4; pp. 727 - 735
• Main Authors: Tham, S.J., National Cheng Kung Univ., Tainan (Taiwan), Chang, C.D, Huang, H.J, Lee, Y.F, Huang, T.S, Chang, C.C
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 2010; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: Acetates; ACID PHOSPHATASE; Acid Phosphatase - metabolism; BIOCHEMISTRY; BIOCHIMIE; Biological and medical sciences; BIOQUIMICA; Buffers; Enzymes; Escherichia coli - metabolism; FOSFATASA ACIDA; Fundamental and applied biological sciences. Psychology; GENE; GENES; GENETICALLY MODIFIED MICROORGANISMS; http://www.fao.org/aos/agrovoc#c_24092; http://www.fao.org/aos/agrovoc#c_27583; http://www.fao.org/aos/agrovoc#c_3214; http://www.fao.org/aos/agrovoc#c_34287; http://www.fao.org/aos/agrovoc#c_36920; http://www.fao.org/aos/agrovoc#c_7657; http://www.fao.org/aos/agrovoc#c_88; http://www.fao.org/aos/agrovoc#c_910; MICRO-ORGANISME MODIFIE GENETIQUE; MICRO-ORGANISME THERMOPHILE; MICROORGANISMOS MODIFICADOS GENETIC; MICROORGANISMOS TERMOFILOS; Nitrophenols; NUCLEOTIDE SEQUENCE; Optimization; Organophosphorus Compounds - metabolism; PHOSPHATASE ACIDE; Phosphoric Monoester Hydrolases - metabolism; PROTEINAS RECOMBINANTES; PROTEINE RECOMBINANTE; Recombinant; recombinant phosphatase; RECOMBINANT PROTEINS; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; Sodium; Substrate Specificity; Tartrates; TEMPERATURA; TEMPERATURE; THERMOPHILIC MICROORGANISMS; Thermus thermophilus; Thermus thermophilus - enzymology; Thermus thermophilus - metabolism; Characterization; Thermus thermophilus; Enzyme; Phosphoric monoester hydrolases; Bacteria; Hydrolases; recombinant phosphatase; Esterases; Acid phosphatase
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.90773

A recombinant putative acid phosphatase from Thermus thermophilus was expressed and purified from Escherichia coli. The recombinant phosphatase displayed activities in a broad range of temperature, from 40 to 90 °C, with optimal temperature at 70 °C. In addition, the recombinant enzyme had activities in a wide range of pH, from 3.6 to 9.1, with optimal pH at 6 in acetate buffer and with optimal pH at 6.5 in Hepes buffer. Furthermore, it showed significant thermal stability and still possessed 44% residual activity after 70 °C treatment for 15 min. Moreover, the recombinant phosphatase showed broad substrates specificities for monophosphate esters, p-nitrophenyl phosphate (pNPP) being the most preferred substrate, and it was able to resist inhibition by sodium tartrate. Additionally, the recombinant protein formed stable oligomer under partially denatured conditions and required calcium ions for enzymic activity.