Biochemical characterization of the δ-carbonic anhydrase from the marine diatom Thalassiosira weissflogii, TweCA.
Diatom genome sequences clearly reveal the presence of different systems for HCO3(-) uptake. Carbon-concentrating mechanisms (CCM) based on HCO3(-) transport and a plastid-localized carbonic anhydrase (CA, EC 4.2.1.1) appear to be more probable than the others because CAs have been identified in the genome of many diatoms. CAs are key enzymes involved in the acquisition of inorganic carbon for photosynthesis in phytoplankton, as they catalyze efficiently the interconversion between carbon dioxide and bicarbonate. Five genetically distinct classes of CAs exist, α-, β-, γ-, δ- and ζ and all of them are metalloenzymes. Recently we investigated for the first time the catalytic activity and inhibition of the δ-class CA from the marine diatom Thalassiosira weissflogii, named TweCA. This enzyme is an efficient catalyst for the CO2 hydration and its inhibition profile with sulfonamide/sulfamate and anions have also been investigated. Here, we report the detailed biochemical characterization and chemico-physical properties of the δ-CA of T. weissflogii. The δ-CA encoding gene was cloned and expressed in Artic Express cells and the recombinant protein purified to homogeneity. Interesting to note that TweCA has no intrinsic esterase activity with 4-nitrophenyl acetate (pNpA) as substrate although the phylogenetic analysis showed that δ-CAs are closer to the α-CAs than to the other classes of such enzymes.
Istituto di Biochimica delle Proteine (IBP) - CNR , Napoli , Italy .
Sonia
S
Del Prete
Daniela
D
Vullo
Viviana
V
De Luca
Claudiu T
CT
Supuran
Clemente
C
Capasso
D000595:Amino Acid Sequence; D001639:Bicarbonates / Q000737:chemistry / Q000378:metabolism; D002245:Carbon Dioxide / Q000737:chemistry / Q000378:metabolism; D002257:Carbonic Anhydrase Inhibitors / Q000737:chemistry*; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000235:genetics; D017377:Diatoms / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics; D016415:Sequence Alignment; D013449:Sulfonamides / Q000737:chemistry*
D001639:Bicarbonates; D002257:Carbonic Anhydrase Inhibitors; D011994:Recombinant Proteins; D013449:Sulfonamides; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases
CO2 hydration; diatoms; esterase activity; metalloenzyme; phylogenetic analysis; δ-Carbonic anhydrase