title: A novel phospholipase A2/esterase from hyperthermophilic archaeon Aeropyrum pernix K1.
abstract: An open reading frame of the hyperthermophilic archaeon Aeropyrum pernix K1 APE2325, which composed of 474 bases, was cloned and expressed in Escherichia coli BL21 (DE3) Codon Plus-RIL. The recombinant protein was purified by Ni-chelation affinity chromatography. It showed a single band with a molecular mass of 18kDa in SDS-PAGE. The purified enzyme exhibited both phospholipase A(2) and esterase activities with the optimal catalytic temperature at 90 degrees C. The enzyme activity was Ca(2+)-independent. Kinetic analysis revealed its Km, k cat, and Vm for the p-nitrophenyl propionate substrate were 103microM, 39s(-1), and 249micromol/min/mg, respectively. The recombinant protein was thermostable and its half-life at 100 degrees C was about 1h.
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun 130023, PR China. yfeng@mail.jlu.edu.cn
authors.forename: Baijing
authors.initials: B
authors.lastname: Wang
authors.forename: Dongmei
authors.initials: D
authors.lastname: Lu
authors.forename: Renjun
authors.initials: R
authors.lastname: Gao
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yang
authors.forename: Shugui
authors.initials: S
authors.lastname: Cao
authors.forename: Yan
authors.initials: Y
authors.lastname: Feng
mesh_terms: D041522:Aeropyrum / Q000201:enzymology*; D000595:Amino Acid Sequence; D004492:Edetic Acid; D004795:Enzyme Stability; D004950:Esterases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008670:Metals; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010741:Phospholipases A / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D054467:Phospholipases A2; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D008670:Metals; D011994:Recombinant Proteins; D004492:Edetic Acid; D004950:Esterases; D010741:Phospholipases A; D054467:Phospholipases A2