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Biosynthesis of Azorhizobium caulinodans Nod factors: study of the activity of the NodABCS proteins by expression of the genes in Escherichia coli

(1995) JOURNAL OF BIOLOGICAL CHEMISTRY. 270(49). p.29217-29223
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Abstract
By in vitro and in vivo studies with Escherichia coil expressing different combinations of the nodABCS genes of Azorhizobium caulinodans, Nod factor intermediates were identified and their structures determined using mass spectrometry. Substrate-product relationships were studied by time course experiments, and the Nod factor biosynthetic pathway was partially resolved. E. coil strains, harboring nodA and/or nocB, did not produce Nod metabolites, whereas the strain expressing nodC produced chitooligosaccharides. Thus, the first committed step was the production of the carbohydrate backbone. Bacitracin and tunicamycin did not affect this step, suggesting that undecaprenyl pyrophosphate-linked intermediates were not involved. The second step was the deacetylation of chitooligosaccharides by NodB since the E. coil strain expressing nodBC produced chitooligosaccharides, deacetylated at the non-reducing end and since the NodC products were precursors of the NodBC products. A strain expressing nodBCS produced N-methylated oligosaccharides, whereas a strain expressing nodCS produced unmethylated oligosaccharides. Time course experiments showed that methylation occurred after deacetylation. Thus, NodS acted after NodB. The NodBCS metabolites were partially converted to Lipo-chitooligosaccharides when the nodABCS genes were expressed, showing that NodA was involved in the acylation and acted after NodS.
Keywords
NODULATION, LOCUS-1, STREPTOCOCCUS, IDENTIFICATION, ENTEROBACTERIAL COMMON ANTIGEN, CHITIN, SEQUENCE

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Chicago
Mergaert, Peter, Wim D’Haeze, Danny Geelen, Danielle Promé, Marc Van Montagu, Roberto Geremia, Jean-Claude Promé, and Marcella Holsters. 1995. “Biosynthesis of Azorhizobium Caulinodans Nod Factors: Study of the Activity of the NodABCS Proteins by Expression of the Genes in Escherichia Coli.” Journal of Biological Chemistry 270 (49): 29217–29223.
APA
Mergaert, P., D’Haeze, W., Geelen, D., Promé, D., Van Montagu, M., Geremia, R., Promé, J.-C., et al. (1995). Biosynthesis of Azorhizobium caulinodans Nod factors: study of the activity of the NodABCS proteins by expression of the genes in Escherichia coli. JOURNAL OF BIOLOGICAL CHEMISTRY, 270(49), 29217–29223.
Vancouver
1.
Mergaert P, D’Haeze W, Geelen D, Promé D, Van Montagu M, Geremia R, et al. Biosynthesis of Azorhizobium caulinodans Nod factors: study of the activity of the NodABCS proteins by expression of the genes in Escherichia coli. JOURNAL OF BIOLOGICAL CHEMISTRY. 1995;270(49):29217–23.
MLA
Mergaert, Peter, Wim D’Haeze, Danny Geelen, et al. “Biosynthesis of Azorhizobium Caulinodans Nod Factors: Study of the Activity of the NodABCS Proteins by Expression of the Genes in Escherichia Coli.” JOURNAL OF BIOLOGICAL CHEMISTRY 270.49 (1995): 29217–29223. Print.
@article{196222,
  abstract     = {By in vitro and in vivo studies with Escherichia coil expressing different combinations of the nodABCS genes of Azorhizobium caulinodans, Nod factor intermediates were identified and their structures determined using mass spectrometry. Substrate-product relationships were studied by time course experiments, and the Nod factor biosynthetic pathway was partially resolved. E. coil strains, harboring nodA and/or nocB, did not produce Nod metabolites, whereas the strain expressing nodC produced chitooligosaccharides. Thus, the first committed step was the production of the carbohydrate backbone. Bacitracin and tunicamycin did not affect this step, suggesting that undecaprenyl pyrophosphate-linked intermediates were not involved. The second step was the deacetylation of chitooligosaccharides by NodB since the E. coil strain expressing nodBC produced chitooligosaccharides, deacetylated at the non-reducing end and since the NodC products were precursors of the NodBC products. A strain expressing nodBCS produced N-methylated oligosaccharides, whereas a strain expressing nodCS produced unmethylated oligosaccharides. Time course experiments showed that methylation occurred after deacetylation. Thus, NodS acted after NodB. The NodBCS metabolites were partially converted to Lipo-chitooligosaccharides when the nodABCS genes were expressed, showing that NodA was involved in the acylation and acted after NodS.},
  author       = {Mergaert, Peter and D'Haeze, Wim and Geelen, Danny and Prom{\'e}, Danielle and Van Montagu, Marc and Geremia, Roberto and Prom{\'e}, Jean-Claude and Holsters, Marcella},
  issn         = {0021-9258},
  journal      = {JOURNAL OF BIOLOGICAL CHEMISTRY},
  keyword      = {NODULATION,LOCUS-1,STREPTOCOCCUS,IDENTIFICATION,ENTEROBACTERIAL COMMON ANTIGEN,CHITIN,SEQUENCE},
  language     = {eng},
  number       = {49},
  pages        = {29217--29223},
  title        = {Biosynthesis of Azorhizobium caulinodans Nod factors: study of the activity of the NodABCS proteins by expression of the genes in Escherichia coli},
  url          = {http://dx.doi.org/10.1074/jbc.270.49.29217},
  volume       = {270},
  year         = {1995},
}

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