Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus

Bacillus sphaericus, a bacterium of biotechnological interest due to its ability to produce mosquitocidal toxins, is unable to use sugars as carbon source. However, ptsHI genes encoding HPr and EI proteins belonging to a PTS were cloned, sequenced and characterized. Both HPr and EI proteins were ful...

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Autores principales:	Alice, A.F., Pérez-Martínez, G., Sánchez-Rivas, C.
Formato:	JOUR
Materias:	6 phosphofructokinase bacterial toxin carbon carrier protein deaminase glycine histidine n acetylglucosamine phosphoenolpyruvate sugar phosphotransferase phosphotransferase serine streptozocin sugar animal cell Bacillus sphaericus Bacillus subtilis bacterial strain bacterium mutant biotechnology carbon source catabolism controlled study energy resource enzyme activity enzyme metabolism gene sequence gene structure genetic code genetic complementation genetic transcription glycolysis in vitro study insecticidal activity molecular cloning mosquito nonhuman nucleotide sequence priority journal protein function protein phosphorylation protein purification review Staphylococcus aureus toxin synthesis transport kinetics wild type Animalia Bacteria (microorganisms) Posibacteria Prokaryota
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_13500872_v149_n7_p1687_Alice
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_13500872_v149_n7_p1687_Alice2023-10-03T16:09:52Z Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus Alice, A.F. Pérez-Martínez, G. Sánchez-Rivas, C. 6 phosphofructokinase bacterial toxin carbon carrier protein deaminase glycine histidine n acetylglucosamine phosphoenolpyruvate sugar phosphotransferase phosphotransferase serine streptozocin sugar animal cell Bacillus sphaericus Bacillus subtilis bacterial strain bacterium mutant biotechnology carbon source catabolism controlled study energy resource enzyme activity enzyme metabolism gene sequence gene structure genetic code genetic complementation genetic transcription glycolysis in vitro study insecticidal activity molecular cloning mosquito nonhuman nucleotide sequence priority journal protein function protein phosphorylation protein purification review Staphylococcus aureus toxin synthesis transport kinetics wild type Animalia Bacillus sphaericus Bacillus sphaericus Bacillus subtilis Bacteria (microorganisms) Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus Bacillus sphaericus, a bacterium of biotechnological interest due to its ability to produce mosquitocidal toxins, is unable to use sugars as carbon source. However, ptsHI genes encoding HPr and EI proteins belonging to a PTS were cloned, sequenced and characterized. Both HPr and EI proteins were fully functional for phosphoenolpyruvate-dependent transphosphorylation in complementation assays using extracts from Staphylococcus aureus mutants for one of these proteins. HPr(His6) was purified from wild-type and a Ser46/Gln mutant of B. sphaericus, and used for in vitro phosphorylation experiments using extracts from either B. sphaericus or Bacillus subtilis as kinase source. The results showed that both phosphorylated forms, P-Ser46-HPr and P-His15-HPr, could be obtained. The findings also proved indirectly the existence of an HPr kinase activity in B. sphaericus. The genetic structure of these ptsHI genes has some unusual features, as they are co-transcribed with genes encoding metabolic enzymes related to N-acetylglucosamine (GlcNAc) catabolism (nagA, nagB and an undetermined orf2). In fact, this bacterium was able to utilize this amino sugar as carbon and energy source, but a ptsH null mutant had lost this characteristic. Investigation of GlcNAc uptake and streptozotocin inhibition in both a wild-type and a ptsH null mutant strain led to the proposal that GlcNAc is transported and phosphorylated by an EIINag element of the PTS, as yet uncharacterized. In addition, GlcNAc-6-phosphate deacetylase and GlcN-6-phosphate deaminase activities were determined; both were induced in the presence of GlcNAc. These results, together with the authors' recent findings of the presence of a phosphofructokinase activity, are strongly indicative of a glycolytic pathway in B. sphaericus. They also open new possibilities for genetic improvements in industrial applications. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_13500872_v149_n7_p1687_Alice
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	6 phosphofructokinase bacterial toxin carbon carrier protein deaminase glycine histidine n acetylglucosamine phosphoenolpyruvate sugar phosphotransferase phosphotransferase serine streptozocin sugar animal cell Bacillus sphaericus Bacillus subtilis bacterial strain bacterium mutant biotechnology carbon source catabolism controlled study energy resource enzyme activity enzyme metabolism gene sequence gene structure genetic code genetic complementation genetic transcription glycolysis in vitro study insecticidal activity molecular cloning mosquito nonhuman nucleotide sequence priority journal protein function protein phosphorylation protein purification review Staphylococcus aureus toxin synthesis transport kinetics wild type Animalia Bacillus sphaericus Bacillus sphaericus Bacillus subtilis Bacteria (microorganisms) Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus
spellingShingle	6 phosphofructokinase bacterial toxin carbon carrier protein deaminase glycine histidine n acetylglucosamine phosphoenolpyruvate sugar phosphotransferase phosphotransferase serine streptozocin sugar animal cell Bacillus sphaericus Bacillus subtilis bacterial strain bacterium mutant biotechnology carbon source catabolism controlled study energy resource enzyme activity enzyme metabolism gene sequence gene structure genetic code genetic complementation genetic transcription glycolysis in vitro study insecticidal activity molecular cloning mosquito nonhuman nucleotide sequence priority journal protein function protein phosphorylation protein purification review Staphylococcus aureus toxin synthesis transport kinetics wild type Animalia Bacillus sphaericus Bacillus sphaericus Bacillus subtilis Bacteria (microorganisms) Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus Alice, A.F. Pérez-Martínez, G. Sánchez-Rivas, C. Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus
topic_facet	6 phosphofructokinase bacterial toxin carbon carrier protein deaminase glycine histidine n acetylglucosamine phosphoenolpyruvate sugar phosphotransferase phosphotransferase serine streptozocin sugar animal cell Bacillus sphaericus Bacillus subtilis bacterial strain bacterium mutant biotechnology carbon source catabolism controlled study energy resource enzyme activity enzyme metabolism gene sequence gene structure genetic code genetic complementation genetic transcription glycolysis in vitro study insecticidal activity molecular cloning mosquito nonhuman nucleotide sequence priority journal protein function protein phosphorylation protein purification review Staphylococcus aureus toxin synthesis transport kinetics wild type Animalia Bacillus sphaericus Bacillus sphaericus Bacillus subtilis Bacteria (microorganisms) Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus
description	Bacillus sphaericus, a bacterium of biotechnological interest due to its ability to produce mosquitocidal toxins, is unable to use sugars as carbon source. However, ptsHI genes encoding HPr and EI proteins belonging to a PTS were cloned, sequenced and characterized. Both HPr and EI proteins were fully functional for phosphoenolpyruvate-dependent transphosphorylation in complementation assays using extracts from Staphylococcus aureus mutants for one of these proteins. HPr(His6) was purified from wild-type and a Ser46/Gln mutant of B. sphaericus, and used for in vitro phosphorylation experiments using extracts from either B. sphaericus or Bacillus subtilis as kinase source. The results showed that both phosphorylated forms, P-Ser46-HPr and P-His15-HPr, could be obtained. The findings also proved indirectly the existence of an HPr kinase activity in B. sphaericus. The genetic structure of these ptsHI genes has some unusual features, as they are co-transcribed with genes encoding metabolic enzymes related to N-acetylglucosamine (GlcNAc) catabolism (nagA, nagB and an undetermined orf2). In fact, this bacterium was able to utilize this amino sugar as carbon and energy source, but a ptsH null mutant had lost this characteristic. Investigation of GlcNAc uptake and streptozotocin inhibition in both a wild-type and a ptsH null mutant strain led to the proposal that GlcNAc is transported and phosphorylated by an EIINag element of the PTS, as yet uncharacterized. In addition, GlcNAc-6-phosphate deacetylase and GlcN-6-phosphate deaminase activities were determined; both were induced in the presence of GlcNAc. These results, together with the authors' recent findings of the presence of a phosphofructokinase activity, are strongly indicative of a glycolytic pathway in B. sphaericus. They also open new possibilities for genetic improvements in industrial applications.
format	JOUR
author	Alice, A.F. Pérez-Martínez, G. Sánchez-Rivas, C.
author_facet	Alice, A.F. Pérez-Martínez, G. Sánchez-Rivas, C.
author_sort	Alice, A.F.
title	Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus
title_short	Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus
title_full	Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus
title_fullStr	Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus
title_full_unstemmed	Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus
title_sort	phosphoenolpyruvate phosphotransferase system and n-acetylglucosamine metabolism in bacillus sphaericus
url	http://hdl.handle.net/20.500.12110/paper_13500872_v149_n7_p1687_Alice
work_keys_str_mv	AT aliceaf phosphoenolpyruvatephosphotransferasesystemandnacetylglucosaminemetabolisminbacillussphaericus AT perezmartinezg phosphoenolpyruvatephosphotransferasesystemandnacetylglucosaminemetabolisminbacillussphaericus AT sanchezrivasc phosphoenolpyruvatephosphotransferasesystemandnacetylglucosaminemetabolisminbacillussphaericus
_version_	1807316789087961088

Phosphoenolpyruvate phosphotransferase system and N-acetylglucosamine metabolism in Bacillus sphaericus

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