Structure of O-specific polysaccharide of Oligotropha carboxidovorans OM5 - a wastewater bacterium

Komaniecka, Iwona; Choma, Adam; Zamlynska, Katarzyna; Sroka-Bartnicka, Anna; Sowiński, Paweł

doi:10.1016/j.carres.2017.01.001

Cited by 11 publications

(6 citation statements)

References 31 publications

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“…A similar polysaccharide of 11 units of α-1,4-linked 3-O-methyl-Dmannose has been identified in Streptomyces griseus (15,16) but in this unique structure some units were acetylated at O-6 positions, a feature common to MGLP. Recently, the wastewater bacterium Oligotropha carboxidovorans was also found to synthesize a related polysaccharide of 35-40 3-O-methyl-D-mannose units, but in this case assembled with α-1,2 linkages (17).…”

Section: Significancementioning

confidence: 99%

Biosynthesis of mycobacterial methylmannose polysaccharides requires a unique 1- O -methyltransferase specific for 3- O -methylated mannosides

Ripoll‐Rozada

Costa

Manso

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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Mycobacteria are a wide group of organisms that includes strict pathogens, such as Mycobacterium tuberculosis, as well as environmental species known as nontuberculous mycobacteria (NTM), some of which—namely Mycobacterium avium—are important opportunistic pathogens. In addition to a distinctive cell envelope mediating critical interactions with the host immune system and largely responsible for their formidable resistance to antimicrobials, mycobacteria synthesize rare intracellular polymethylated polysaccharides implicated in the modulation of fatty acid metabolism, thus critical players in cell envelope assembly. These are the 6-O-methylglucose lipopolysaccharides (MGLP) ubiquitously detected across the Mycobacterium genus, and the 3-O-methylmannose polysaccharides (MMP) identified only in NTM. The polymethylated nature of these polysaccharides renders the intervening methyltransferases essential for their optimal function. Although the knowledge of MGLP biogenesis is greater than that of MMP biosynthesis, the methyltransferases of both pathways remain uncharacterized. Here, we report the identification and characterization of a unique S-adenosyl-l-methionine–dependent sugar 1-O-methyltransferase (MeT1) from Mycobacterium hassiacum that specifically blocks the 1-OH position of 3,3′-di-O-methyl-4α-mannobiose, a probable early precursor of MMP, which we chemically synthesized. The high-resolution 3D structure of MeT1 in complex with its exhausted cofactor, S-adenosyl-l-homocysteine, together with mutagenesis studies and molecular docking simulations, unveiled the enzyme’s reaction mechanism. The functional and structural properties of this unique sugar methyltransferase further our knowledge of MMP biosynthesis and provide important tools to dissect the role of MMP in NTM physiology and resilience.

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Section: Significancementioning

confidence: 99%

Biosynthesis of mycobacterial methylmannose polysaccharides requires a unique 1- O -methyltransferase specific for 3- O -methylated mannosides

Ripoll‐Rozada

Costa

Manso

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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“…LPS was isolated from previously delipidated and enzymatically digested cells, using the hot 45% phenol/water extraction method [17,18], as described by Komaniecka and co-workers [29]. The LPS was obtained after ultracentrifugation (104,000× g, 4 h, 4 • C).…”

Section: Lps and Lipid A Isolation And Purificationmentioning

confidence: 99%

Lipid A from Oligotropha carboxidovorans Lipopolysaccharide That Contains Two Galacturonic Acid Residues in the Backbone and Malic Acid A Tertiary Acyl Substituent

Choma

Zamlynska

Mazur

et al. 2020

IJMS

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The free-living Gram-negative bacterium Oligotropha carboxidovorans (formerly: Pseudomonas carboxydovorans), isolated from wastewater, is able to live in aerobic and, facultatively, in autotrophic conditions, utilizing carbon monoxide or hydrogen as a source of energy. The structure of O. carboxidovorans lipid A, a hydrophobic part of lipopolysaccharide, was studied using NMR spectroscopy and high-resolution mass spectrometry (MALDI-ToF MS) techniques. It was demonstrated that the lipid A backbone is composed of two d-GlcpN3N residues connected by a β-(1→6) glycosidic linkage, substituted by galacturonic acids (d-GalpA) at C-1 and C-4’ positions. Both diaminosugars are symmetrically substituted by 3-hydroxy fatty acids (12:0(3-OH) and 18:0(3-OH)). Ester-linked secondary acyl residues (i.e., 18:0, and 26:0(25-OH) and a small amount of 28:0(27-OH)) are located in the distal part of lipid A. These very long-chain hydroxylated fatty acids (VLCFAs) were found to be almost totally esterified at the (ω-1)-OH position with malic acid. Similarities between the lipid A of O. carboxidovorans and Mesorhizobium loti, Rhizobium leguminosarum, Caulobacter crescentus as well as Aquifex pyrophylus were observed and discussed from the perspective of the genomic context of these bacteria.

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“…The α-(1→4) linkage between mannoses found in MMP appears to be extremely rare in nature 13 , which renders the key α-(1→4)-mannosyltransferase a functionally and structurally unique glycosyltransferase. Indeed, mannans from plants show a clear prevalence of β-(1→4) linkages, whereas those from fungi and bacteria essentially contain α-(1→2), α-(1→3), and α-(1→6) linkages and branching 14 – 16 . Also, in the mycobacterial mannose polymers phosphatidyl- myo -inositol mannosides (PIM), lipomannan (LM) and lipoarabinomannan (LAM), mannose units are essentially connected through α-(1→6) and α-(1→2) glycosidic bonds 7 .…”

Section: Introductionmentioning

confidence: 99%

Self-recycling and partially conservative replication of mycobacterial methylmannose polysaccharides

et al. 2023

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The steep increase in nontuberculous mycobacteria (NTM) infections makes understanding their unique physiology an urgent health priority. NTM synthesize two polysaccharides proposed to modulate fatty acid metabolism: the ubiquitous 6-O-methylglucose lipopolysaccharide, and the 3-O-methylmannose polysaccharide (MMP) so far detected in rapidly growing mycobacteria. The recent identification of a unique MMP methyltransferase implicated the adjacent genes in MMP biosynthesis. We report a wide distribution of this gene cluster in NTM, including slowly growing mycobacteria such as Mycobacterium avium, which we reveal to produce MMP. Using a combination of MMP purification and chemoenzymatic syntheses of intermediates, we identified the biosynthetic mechanism of MMP, relying on two enzymes that we characterized biochemically and structurally: a previously undescribed α–endomannosidase that hydrolyses MMP into defined-sized mannoligosaccharides that prime the elongation of new daughter MMP chains by a rare α-(1→4)-mannosyltransferase. Therefore, MMP biogenesis occurs through a partially conservative replication mechanism, whose disruption affected mycobacterial growth rate at low temperature.

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Structure of O-specific polysaccharide of Oligotropha carboxidovorans OM5 - a wastewater bacterium

Cited by 11 publications

References 31 publications

Biosynthesis of mycobacterial methylmannose polysaccharides requires a unique 1- O -methyltransferase specific for 3- O -methylated mannosides

Biosynthesis of mycobacterial methylmannose polysaccharides requires a unique 1- O -methyltransferase specific for 3- O -methylated mannosides

Lipid A from Oligotropha carboxidovorans Lipopolysaccharide That Contains Two Galacturonic Acid Residues in the Backbone and Malic Acid A Tertiary Acyl Substituent

Self-recycling and partially conservative replication of mycobacterial methylmannose polysaccharides

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