Acquisition of a new type of fructose-1,6-bisphosphatase with resistance to hydrogen peroxide in cyanobacteria: molecular characterization of the enzyme from Synechocystis PCC 6803

Tamoi, Masahiro; Murakami, Akiko; Takeda, Toru; Shigeoka, Shigeru

doi:10.1016/s0167-4838(97)00208-2

Cited by 64 publications

(47 citation statements)

References 41 publications

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“…Bacterial FBPase are bifunctional enzymes with both FBPase and SBPase activities. The class II variants typically have high SBPase-to-FBPase ratios (34) and play a role in one variant of the rearrangement phase of the RuMP pathway.…”

Section: Resultsmentioning

confidence: 99%

Plasmid-Dependent Methylotrophy in Thermotolerant Bacillus methanolicus

et al. 2004

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Bacillus methanolicus can efficiently utilize methanol as a sole carbon source and has an optimum growth temperature of 50°C. With the exception of mannitol, no sugars have been reported to support rapid growth of this organism, which is classified as a restrictive methylotroph. Here we describe the DNA sequence and characterization of a 19,167-bp circular plasmid, designated pBM19, isolated from B. methanolicus MGA3. Sequence analysis of pBM19 demonstrated the presence of the methanol dehydrogenase gene, mdh, which is crucial for methanol consumption in this bacterium. In addition, five genes (pfk, encoding phosphofructokinase; rpe, encoding ribulose-5-phosphate 3-epimerase; tkt, encoding transketolase; glpX, encoding fructose-1,6-bisphosphatase; and fba, encoding fructose-1,6-bisphosphate aldolase) with deduced roles in methanol assimilation via the ribulose monophosphate pathway are encoded by pBM19. A shuttle vector, pTB1.9, harboring the pBM19 minimal replicon (repB and ori) was constructed and used to transform MGA3. Analysis of the resulting recombinant strain demonstrated that it was cured of pBM19 and was not able to grow on methanol. A pTB1.9 derivative harboring the complete mdh gene could not restore growth on methanol when it was introduced into the pBM19-cured strain, suggesting that additional pBM19 genes are required for consumption of this carbon source. Screening of 13 thermotolerant B. methanolicus wild-type strains showed that they all harbor plasmids similar to pBM19, and this is the first report describing plasmid-linked methylotrophy in any microorganism. Our findings should have an effect on future genetic manipulations of this organism, and they contribute to a new understanding of the biology of methylotrophs.

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Section: Resultsmentioning

confidence: 99%

Plasmid-Dependent Methylotrophy in Thermotolerant Bacillus methanolicus

et al. 2004

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“…Primary sequence alignment with biochemically characterized class II FBPases, including Synechocystis sp. PCC6803 Fbp1 (25), E. coli GlpX (16), Mycobacterium tuberculosis H37Rv Rv1099c (50), and C. glutamicum Fbp (42), revealed Ͼ50 conserved amino acid residues (Fig. 2).…”

Section: Bioinformatic Analysis and Phylogeny Of The Fbpases Glpxmentioning

confidence: 99%

“…FBPase II is present in Escherichia coli, encoded by glpX, and in Synechocystis sp. strain PCC6803 (25). FBPase III is present, e.g., in Bacillus subtilis (encoded by fbp) (26), and FBPase IV is present in Pyrococcus furiosus (encoded by fbpA) (27), Methanococcus jannaschii (28), and Archaeoglobus fulgidus (29).…”

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confidence: 99%

Characterization of Fructose 1,6-Bisphosphatase and Sedoheptulose 1,7-Bisphosphatase from the Facultative Ribulose Monophosphate Cycle Methylotroph Bacillus methanolicus

Stolzenberger¹,

Lindner

Persicke

et al. 2013

J Bacteriol

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The genome of the facultative ribulose monophosphate (RuMP) cycle methylotroph Bacillus methanolicus encodes two bisphosphatases (GlpX), one on the chromosome (GlpX C ) and one on plasmid pBM19 (GlpX P ), which is required for methylotrophy. Both enzymes were purified from recombinant Escherichia coli and were shown to be active as fructose 1,6-bisphosphatases (FBPases). The FBPase-negative Corynebacterium glutamicum ⌬fbp mutant could be phenotypically complemented with glpX C and glpX P from B. methanolicus. GlpX P and GlpX C share similar functional properties, as they were found here to be active as homotetramers in vitro, activated by Mn 2؉ ions and inhibited by Li ؉ , but differed in terms of the kinetic parameters. GlpX C showed a much higher catalytic efficiency and a lower K m for fructose 1,6-bisphosphate (86.3 s ؊1 mM ؊1 and 14 ؎ 0.5 M, respectively) than GlpX P (8.8 s ؊1 mM ؊1 and 440 ؎ 7.6 M, respectively), indicating that GlpX C is the major FBPase of B. methanolicus. Both enzymes were tested for activity as sedoheptulose 1,7-bisphosphatase (SBPase), since a SBPase variant of the ribulose monophosphate cycle has been proposed for B. methanolicus. The substrate for the SBPase reaction, sedoheptulose 1,7-bisphosphate, could be synthesized in vitro by using both fructose 1,6-bisphosphate aldolase proteins from B. methanolicus. Evidence for activity as an SBPase could be obtained for GlpX P but not for GlpX C . Based on these in vitro data, GlpX P is a promiscuous SBPase/FBPase and might function in the RuMP cycle of B. methanolicus.

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“…The type I FBPase is the most widely distributed among living organisms and is the primary FBPase in Esche-richia coli, most bacteria, a few archaea, and all eukaryotes (9,(11)(12)(13)(14)(15). The type II FBPases are represented by the E. coli GlpX and FBPase F-I from Synechocystis PCC6803 (9,16); type III is represented by the Bacillus subtilis FBPase (17); type IV is represented by the dual activity FBPases/inosine monophosphatases FbpA from Pyrococcus furiosus (18), MJ0109 from Methanococcus jannaschii (19), and AF2372 from Archaeoglobus fulgidus (20); and type V is represented by the FBPases TK2164 from Pyrococcus (Thermococcus) kodakaraensis and ST0318 from Sulfolobus tokodai (10,21).…”

mentioning

confidence: 99%

Structural and Biochemical Characterization of the Type II Fructose-1,6-bisphosphatase GlpX from Escherichia coli

Brown

Singer

Лунин

et al. 2009

Journal of Biological Chemistry

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Gluconeogenesis is an important metabolic pathway, which produces glucose from noncarbohydrate precursors such as organic acids, fatty acids, amino acids, or glycerol. Fructose-1,6-bisphosphatase, a key enzyme of gluconeogenesis, is found in all organisms, and five different classes of these enzymes have been identified. Here we demonstrate that Escherichia coli has two class II fructose-1,6-bisphosphatases, GlpX and YggF, which show different catalytic properties. We present the first crystal structure of a class II fructose-1,6-bisphosphatase (GlpX) determined in a free state and in the complex with a substrate (fructose 1,6-bisphosphate) or inhibitor (phosphate). The crystal structure of the ligand-free GlpX revealed a compact, globular shape with two ␣/␤-sandwich domains. The core fold of GlpX is structurally similar to that of Li ؉ -sensitive phosphatases implying that they have a common evolutionary origin and catalytic mechanism. The structure of the GlpX complex with fructose 1,6-bisphosphate revealed that the active site is located between two domains and accommodates several conserved residues coordinating two metal ions and the substrate. The third metal ion is bound to phosphate 6 of the substrate. Inorganic phosphate strongly inhibited activity of both GlpX and YggF, and the crystal structure of the GlpX complex with phosphate demonstrated that the inhibitor molecule binds to the active site. Alanine replacement mutagenesis of GlpX identified 12 conserved residues important for activity and suggested that Thr 90 is the primary catalytic residue. Our data provide insight into the molecular mechanisms of the substrate specificity and catalysis of GlpX and other class II fructose-1,6-bisphosphatases.Fructose-1,6-bisphosphatase (FBPase, 2 EC 3.1.3.11), a key enzyme of gluconeogenesis, catalyzes the hydrolysis of fructose 1,6-bisphosphate to form fructose 6-phosphate and orthophosphate. It is the reverse of the reaction catalyzed by phosphofructokinase in glycolysis, and the product, fructose 6-phosphate, is an important precursor in various biosynthetic pathways (1). In all organisms, gluconeogenesis is an important metabolic pathway that allows the cells to synthesize glucose from noncarbohydrate precursors, such as organic acids, amino acids, and glycerol. FBPases are members of the large superfamily of lithium-sensitive phosphatases, which includes three families of inositol phosphatases and FBPases (the phosphoesterase clan CL0171, 3167 sequences, Pfam data base). These enzymes show metal-dependent and lithium-sensitive phosphomonoesterase activity and include inositol polyphosphate 1-phosphatases, inositol monophosphatases (IMPases), 3Ј-phosphoadenosine 5Ј-phosphatases (PAPases), and enzymes acting on both inositol 1,4-bisphosphate and PAP (PIPases) (2). They possess a common structural core with the active site lying between ␣ϩ␤ and ␣/␤ domains (3). Li ϩ -sensitive phosphatases are putative targets for lithium therapy in the treatment of manic depressive patients (4), whereas FBPases are targets f...

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Acquisition of a new type of fructose-1,6-bisphosphatase with resistance to hydrogen peroxide in cyanobacteria: molecular characterization of the enzyme from Synechocystis PCC 6803

Cited by 64 publications

References 41 publications

Plasmid-Dependent Methylotrophy in Thermotolerant Bacillus methanolicus

Plasmid-Dependent Methylotrophy in Thermotolerant Bacillus methanolicus

Characterization of Fructose 1,6-Bisphosphatase and Sedoheptulose 1,7-Bisphosphatase from the Facultative Ribulose Monophosphate Cycle Methylotroph Bacillus methanolicus

Structural and Biochemical Characterization of the Type II Fructose-1,6-bisphosphatase GlpX from Escherichia coli

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