Ceriporiopsis subvermispora is a selective white-rot fungus that degrades lignin at a site far from the hyphae and extracellular enzymes, without intensive damage to the cellulose. In selective ligninolysis, low molecular mass metabolites play a principal role and amphipathic substances are involved to control the degradation and transport of hydrophobic aromatic molecules, including lignin and lipids; however, secretion of the amphipathic substances by this fungus has not been well understood, except for alk(en)yl itaconates called ceriporic acids, which have a weak amphiphilicity. Herein, we report for the first time that the fungus secretes cerebrosides that are classified as glycosphingolipids. By using liquid chromatography electron spray ionization mass spectrometry (LC-ESI-MS) and nuclear magnetic resonance (NMR) spectroscopy coupled with stable isotope feeding experiments with 13C-glucose and 15N-ammonium sulfate, the cerebrosides were determined to be N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-4E,8E-sphingadienine, N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-4E,8Z-sphingadienine, and N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-9-methyl-4E,8E-sphingadienine. The cerebrosides are strong amphipathic substances and potential metabolites for regulating difference and symbiosis within the microbial community.

glycolipid, cerebroside, Basidiomycete, white-rot fungi

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

CCCCCCCCCCCCCC[C@@H](O)C(=O)NC(CO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)C(O)/C=C/CC/C=C(\C)CCCCCCCCC

728.065 g/mol (C₄₁H₇₇NO₉)

Ceriporiopsis subvermispora is a selective white-rot fungus that degrades lignin at a site far from the hyphae and extracellular enzymes, without intensive damage to the cellulose. In selective ligninolysis, low molecular mass metabolites play a principal role and amphipathic substances are involved to control the degradation and transport of hydrophobic aromatic molecules, including lignin and lipids; however, secretion of the amphipathic substances by this fungus has not been well understood, except for alk(en)yl itaconates called ceriporic acids, which have a weak amphiphilicity. Herein, we report for the first time that the fungus secretes cerebrosides that are classified as glycosphingolipids. By using liquid chromatography electron spray ionization mass spectrometry (LC-ESI-MS) and nuclear magnetic resonance (NMR) spectroscopy coupled with stable isotope feeding experiments with 13C-glucose and 15N-ammonium sulfate, the cerebrosides were determined to be N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-4E,8E-sphingadienine, N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-4E,8Z-sphingadienine, and N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-9-methyl-4E,8E-sphingadienine. The cerebrosides are strong amphipathic substances and potential metabolites for regulating difference and symbiosis within the microbial community.

glycolipid, cerebroside, Basidiomycete, white-rot fungi

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6	C7	C8	C9	C10	C11	C12	C13	C14	C15	C16	C17	C18
1	bDGlcp	104.9	75.0	77.9	71.6	78.1	62.7
2	l?2HOPam	174.2	73.1	36.0	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	24.2	14.5
	xXSphdC18	69.8	54.7	72.9	134.5	131.3	33.7	33.5	132.0	130.8	33.5	?	?	?	?	?	?	?	?


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6	H7	H8	H9	H10	H11	H12	H13	H14	H15	H16	H17	H18
1	bDGlcp	4.24	3.16	3.33	3.25	3.25	3.65-3.84
2	l?2HOPam	-	3.96	1.54-1.68	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.87	0.88
	xXSphdC18	3.69-4.09	3.97	4.12	5.71	5.46	1.96	2.05	5.41	5.42	2.05	?	?	?	?	?	?	?	?


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6	C7/H7	C8/H8	C9/H9	C10/H10	C11/H11	C12/H12	C13/H13	C14/H14	C15/H15	C16/H16	C17/H17	C18/H18
1	bDGlcp	104.9/4.24	75.0/3.16	77.9/3.33	71.6/3.25	78.1/3.25	62.7/3.65-3.84
2	l?2HOPam		73.1/3.96	36.0/1.54-1.68	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	24.2/1.87	14.5/0.88
	xXSphdC18	69.8/3.69-4.09	54.7/3.97	72.9/4.12	134.5/5.71	131.3/5.46	33.7/1.96	33.5/2.05	132.0/5.41	130.8/5.42	33.5/2.05	?/?	?/?	?/?	?/?	?/?	?/?	?/?	?/?

The spectrum also has 8 signals at unknown positions (not plotted).

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

CCCCCCCCC/C=C/CC/C=C/C(O)C(CO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)NC(=O)C(O)CCCCCCCCCCCCCC

714.038 g/mol (C₄₀H₇₅NO₉)

Ceriporiopsis subvermispora is a selective white-rot fungus that degrades lignin at a site far from the hyphae and extracellular enzymes, without intensive damage to the cellulose. In selective ligninolysis, low molecular mass metabolites play a principal role and amphipathic substances are involved to control the degradation and transport of hydrophobic aromatic molecules, including lignin and lipids; however, secretion of the amphipathic substances by this fungus has not been well understood, except for alk(en)yl itaconates called ceriporic acids, which have a weak amphiphilicity. Herein, we report for the first time that the fungus secretes cerebrosides that are classified as glycosphingolipids. By using liquid chromatography electron spray ionization mass spectrometry (LC-ESI-MS) and nuclear magnetic resonance (NMR) spectroscopy coupled with stable isotope feeding experiments with 13C-glucose and 15N-ammonium sulfate, the cerebrosides were determined to be N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-4E,8E-sphingadienine, N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-4E,8Z-sphingadienine, and N-hydroxyoctadecanoyl-1-O-β-D-glucopyranosyl-9-methyl-4E,8E-sphingadienine. The cerebrosides are strong amphipathic substances and potential metabolites for regulating difference and symbiosis within the microbial community.

glycolipid, cerebroside, Basidiomycete, white-rot fungi

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6	C7	C8	C9	C10	C11	C12	C13	C14	C15	C16
1	bDGlcp	104.9	75.0	77.9	71.6	78.1	62.7
2	l?2HOPam	174.2	73.1	36.0	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	30.8	24.2	14.5
	Subst	69.8	54.7	72.9	134.5	131.3	33.7	27.9-28.2	131.3	130.2	27.9-28.2


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6	H7	H8	H9	H10	H11	H12	H13	H14	H15	H16
1	bDGlcp	4.24	3.16	3.33	3.25	3.25	3.65-3.84
2	l?2HOPam	-	3.96	1.54-1.68	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.27	1.87	0.88
	Subst	3.69-4.09	3.97	4.12	5.71	5.46	1.96	2.02-2.10	5.35	5.35	2.02-2.10


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6	C7/H7	C8/H8	C9/H9	C10/H10	C11/H11	C12/H12	C13/H13	C14/H14	C15/H15	C16/H16
1	bDGlcp	104.9/4.24	75.0/3.16	77.9/3.33	71.6/3.25	78.1/3.25	62.7/3.65-3.84
2	l?2HOPam		73.1/3.96	36.0/1.54-1.68	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	30.8/1.27	24.2/1.87	14.5/0.88
	Subst	69.8/3.69-4.09	54.7/3.97	72.9/4.12	134.5/5.71	131.3/5.46	33.7/1.96	27.9-28.2/2.02-2.10	131.3/5.35	130.2/5.35	27.9-28.2/2.02-2.10

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

CCCCCCCCCC/C=C\CC/C=C/C(O)C(CO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)NC(=O)C(O)CCCCCCCCCCCCCC

728.065 g/mol (C₄₁H₇₇NO₉)