PMID- 31766749
OWN - NLM
STAT- MEDLINE
DCOM- 20200420
LR  - 20200420
IS  - 1660-3397 (Electronic)
IS  - 1660-3397 (Linking)
VI  - 17
IP  - 12
DP  - 2019 Nov 22
TI  - A Novel Alkaline Phosphatase/Phosphodiesterase, CamPhoD, from Marine Bacterium 
      Cobetia amphilecti KMM 296.
LID - 10.3390/md17120657 [doi]
LID - 657
AB  - A novel extracellular alkaline phosphatase/phosphodiesterase from the structural 
      protein family PhoD that encoded by the genome sequence of the marine bacterium 
      Cobetia amphilecti KMM 296 (CamPhoD) has been expressed in Escherichia coli 
      cells. The calculated molecular weight, the number of amino acids, and the 
      isoelectric point (pI) of the mature protein's subunit are equal to 54832.98 Da, 
      492, and 5.08, respectively. The salt-tolerant, bimetal-dependent enzyme CamPhoD 
      has a molecular weight of approximately 110 kDa in its native state. CamPhoD is 
      activated by Co(2+), Mg(2+), Ca(2+), or Fe(3+) at a concentration of 2 mM and 
      exhibits maximum activity in the presence of both Co(2+) and Fe(3+) ions in the 
      incubation medium at pH 9.2. The exogenous ions, such as Zn(2+), Cu(2+), and 
      Mn(2+), as well as chelating agents EDTA and EGTA, do not have an appreciable 
      effect on the CamPhoD activity. The temperature optimum for the CamPhoD activity 
      is 45  degrees C. The enzyme catalyzes the cleavage of phosphate mono- and diester bonds 
      in nucleotides, releasing inorganic phosphorus from p-nitrophenyl phosphate 
      (pNPP) and guanosine 5'-triphosphate (GTP), as determined by the Chen method, 
      with rate approximately 150- and 250-fold higher than those of bis-pNPP and 
      5'-pNP-TMP, respectively. The Michaelis-Menten constant (K(m)), V(max), and 
      efficiency (k(cat)/K(m)) of CamPhoD were 4.2 mM, 0.203 mM/min, and 7988.6 
      S(-1)/mM; and 6.71 mM, 0.023 mM/min, and 1133.0 S(-1)/mM for pNPP and bis-pNPP as 
      the chromogenic substrates, respectively. Among the 3D structures currently 
      available, in this study we found only the low identical structure of the 
      Bacillus subtilis enzyme as a homologous template for modeling CamPhoD, with a 
      new architecture of the phosphatase active site containing Fe(3+) and two Ca(2+) 
      ions. It is evident that the marine bacterial phosphatase/phosphidiesterase 
      CamPhoD is a new structural member of the PhoD family.
FAU - Noskova, Yulia
AU  - Noskova Y
AD  - Laboratories of Marine Biochemistry and Bioassays and Mechanisms of Action of 
      Biologically Active Substances, G.B. Elyakov Pacific Institute of Bioorganic 
      Chemistry, Far Eastern Branch, the Russian Academy of Sciences, 690022 
      Vladivostok, Russia.
FAU - Likhatskaya, Galina
AU  - Likhatskaya G
AD  - Laboratories of Marine Biochemistry and Bioassays and Mechanisms of Action of 
      Biologically Active Substances, G.B. Elyakov Pacific Institute of Bioorganic 
      Chemistry, Far Eastern Branch, the Russian Academy of Sciences, 690022 
      Vladivostok, Russia.
FAU - Terentieva, Natalia
AU  - Terentieva N
AD  - Laboratories of Marine Biochemistry and Bioassays and Mechanisms of Action of 
      Biologically Active Substances, G.B. Elyakov Pacific Institute of Bioorganic 
      Chemistry, Far Eastern Branch, the Russian Academy of Sciences, 690022 
      Vladivostok, Russia.
FAU - Son, Oksana
AU  - Son O
AD  - School of Economics and Management of Far East Federal University, 690950 
      Vladivostok, Russia.
FAU - Tekutyeva, Liudmila
AU  - Tekutyeva L
AD  - School of Economics and Management of Far East Federal University, 690950 
      Vladivostok, Russia.
FAU - Balabanova, Larissa
AU  - Balabanova L
AD  - Laboratories of Marine Biochemistry and Bioassays and Mechanisms of Action of 
      Biologically Active Substances, G.B. Elyakov Pacific Institute of Bioorganic 
      Chemistry, Far Eastern Branch, the Russian Academy of Sciences, 690022 
      Vladivostok, Russia.
AD  - School of Economics and Management of Far East Federal University, 690950 
      Vladivostok, Russia.
LA  - eng
PT  - Journal Article
DEP - 20191122
PL  - Switzerland
TA  - Mar Drugs
JT  - Marine drugs
JID - 101213729
RN  - 0 (Recombinant Proteins)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
RN  - EC 3.1.4.1 (Phosphodiesterase I)
RN  - Cobetia amphilecti
SB  - IM
MH  - Alkaline Phosphatase/*chemistry/genetics/isolation & purification/metabolism
MH  - Aquatic Organisms/*enzymology/genetics
MH  - Enzyme Assays
MH  - Halomonadaceae/*enzymology/genetics
MH  - Phosphodiesterase I/*chemistry/genetics/isolation & purification/metabolism
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/chemistry/genetics/isolation & purification/metabolism
PMC - PMC6950083
OTO - NOTNLM
OT  - Cobetia amphilecti
OT  - PhoD
OT  - bimetal-dependent phosphodiesterase
OT  - marine bacterium
OT  - recombinant alkaline phosphatase
COIS- The authors declare no conflict of interest.
EDAT- 2019/11/27 06:00
MHDA- 2020/04/21 06:00
PMCR- 2019/12/01
CRDT- 2019/11/27 06:00
PHST- 2019/10/22 00:00 [received]
PHST- 2019/11/18 00:00 [revised]
PHST- 2019/11/19 00:00 [accepted]
PHST- 2019/11/27 06:00 [entrez]
PHST- 2019/11/27 06:00 [pubmed]
PHST- 2020/04/21 06:00 [medline]
PHST- 2019/12/01 00:00 [pmc-release]
AID - md17120657 [pii]
AID - marinedrugs-17-00657 [pii]
AID - 10.3390/md17120657 [doi]
PST - epublish
SO  - Mar Drugs. 2019 Nov 22;17(12):657. doi: 10.3390/md17120657.