Cloning, sequencing and characterization of the alkaline phosphatase gene (phoD) fromZymomonas mobilis

Abstract

The phoD gene encoding the membrane-bound alkaline phosphatase (ALPI) from Zymomonas mobilis CP4 was cloned and sequenced. Both the translated sequence and the properties of the recombinant enzyme were unusual. Z. mobilis ALPI was monomeric (M(r) 62,926) and hydrolysed nucleotides more effectively than sugar phosphates. The translated sequence contained a single hydrophobic segment near the N-terminus which may serve as a membrane-anchor in Z. mobilis, although the recombinant enzyme was recovered in the cytoplasmic fraction of Escherichia coli. The predicted amino acid sequence for ALPI did not align well with other ALPs or other known genes. However, some similarity to E. coli ALP was noted in the metal-binding and phosphate-binding regions. Two other regions were identified with similarity to the active sites of pyruvate kinase and mammalian 5'-nucleotide phosphodiesterase (also membrane-bound), respectively. It is likely that Z. mobilis phoD represents a new class of alkaline phosphatase genes which has not been described previously.

Purification and characterization of alkaline phosphatase containing phosphotyrosyl phosphatase activity from the bacterium Prevotella intermedia

“…These results strongly indicated that the PiALP would belong to the same family with PhoD from Z. mobilis and PhoV from Synechococcus . On the other hand, in previous studies [9, 10], PTPase activities in these two ALPases were not investigated. Unfortunately, no information is available about PTPase activities of PhoD and PhoV at present.…”

“…A significant high sequence similarity was found between PiALP and the ALPases of Z. mobilis (PhoD) and the cyanobacterium Synechococcus (PhoV). The PiALP contained a putative phosphorylation site (LAVSLSSTD(Y/A)V) that is conserved between PhoD and PhoV, although the involvement of this site in regulation of activity remains speculative [9, 10]. These results strongly indicated that the PiALP would belong to the same family with PhoD from Z. mobilis and PhoV from Synechococcus .…”

“…As shown in Table 4 , a distinct sequence was obtained, Thr‐Asp‐Met‐Leu‐Ala‐Val‐Ser‐Val‐Ser‐Ser‐Thr‐Asp‐Ala‐Ile‐Gly‐His‐Lys‐Tyr‐Gly‐Thr. The amino acid sequence had 65% identity with two ALPases, PhoD from Zymomonas mobilis [9]and PhoV from Synechococcus [10]. Interestingly, a sequence homologous to the putative phosphorylation site [10], Leu‐Ala‐Val‐Ser‐Leu‐Ser‐Ser‐Thr‐Asp‐Ala‐Val, was found.…”

Secreted Phosphatase and Deoxyribonuclease Are Required by Pseudomonas aeruginosa To Defend against Neutrophil Extracellular Traps

“…Interestingly, PhoD phosphatases possess both monoesterase as well as phosphodiesterase (PDase) activities (30,31). Moreover, EddA homologs, such as the Zymomonas mobilis CP4 PhoD alkaline phosphatase, have been shown to hydrolyze nucleotides more effectively than sugar phosphates (33).…”

Cloning and Expression of the Unique Ca2+-ATPase from Flavobacterium odoratum

“…The F. odoratum Ca 2ϩ -ATPase exhibits moderate sequence homology (30%) with an alkaline phosphatase (phoD) from Zymomonas mobilis (23). Despite weak homology to short regions of several Ca 2ϩ -ATPases, including the Synechococcus pacL, which has been hypothesized to be a Ca 2ϩ -ATPase, the Z. mobilis sequence contains none of the highly conserved regions found in other P-type ATPases, including the phosphorylation site.…”