Chapter 4
Lipid and Lipoarabinomannan Isolation
and Characterization
Marie-Antoinette Lane ´ elle, Lucie Spina, Je ´ ro ˆ me Nigou,
Anne Lemassu, and Mamadou Daffe ´
Abstract
The very high content of structurally diverse and biologically active lipids of exotic structures is the hallmark
of Mycobacteria. As such the lipid composition is commonly used to characterize mycobacterial strains at
the species and type-species levels. The present chapter describes the methods that allow the purification of
the most commonly isolated biologically active lipids and those used for analyzing extractable lipids and
their constituents, cell wall-linked mycolic acids (MA), and lipoarabinomannan (LAM). These involve
various chromatographic techniques and analytical procedures necessary for structural and metabolic
studies of mycobacterial lipids. In addition, as the use of physical methods has brought important overhang
on chemical structures of the very-long-chain MA, which typify mycobacteria, NMR and mass spectrometry
data of these specific fatty acids are included.
Key words Lipids, Glycolipids, Mycolic acids, Cell wall, Lipid analysis
1 Introduction
Mycobacteria are extraordinarily rich in lipid, constituting up to
40% of their dry weight. Compared to the lipid-rich cell wall of
diderm Gram-negative bacteria, with some 20%, the mycobacterial
cell walls contain up to 60% of lipids [1]. This high lipid content has
been associated with various other singular properties of mycobacteria, such as their acid fastness, the granuloma formation, and their
impermeability to antibiotics and solutes [2]. Expectedly, the
sequenced mycobacterial genomes contain a high percent of
genes encoding proteins putatively involved in lipid metabolism.
Mycobacteria, like Gram-negative, contain two membranes
[3]: (1) the plasma membrane whose composition is similar to
other membranes, i.e., a bilayer of phospholipids and proteins,
and (2) an outer membrane (mycomembrane), whose inner leaflet
consists of α-branched and β-hydroxylated C60-C90 fatty acyls
called mycolic acyl (MA) chains, covalently attached to the cell
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_4, © Springer Science+Business Media, LLC, part of Springer Nature 2021
109
Lipid and Lipoarabinomannan Isolation
and Characterization
Marie-Antoinette Lane ´ elle, Lucie Spina, Je ´ ro ˆ me Nigou,
Anne Lemassu, and Mamadou Daffe ´
Abstract
The very high content of structurally diverse and biologically active lipids of exotic structures is the hallmark
of Mycobacteria. As such the lipid composition is commonly used to characterize mycobacterial strains at
the species and type-species levels. The present chapter describes the methods that allow the purification of
the most commonly isolated biologically active lipids and those used for analyzing extractable lipids and
their constituents, cell wall-linked mycolic acids (MA), and lipoarabinomannan (LAM). These involve
various chromatographic techniques and analytical procedures necessary for structural and metabolic
studies of mycobacterial lipids. In addition, as the use of physical methods has brought important overhang
on chemical structures of the very-long-chain MA, which typify mycobacteria, NMR and mass spectrometry
data of these specific fatty acids are included.
Key words Lipids, Glycolipids, Mycolic acids, Cell wall, Lipid analysis
1 Introduction
Mycobacteria are extraordinarily rich in lipid, constituting up to
40% of their dry weight. Compared to the lipid-rich cell wall of
diderm Gram-negative bacteria, with some 20%, the mycobacterial
cell walls contain up to 60% of lipids [1]. This high lipid content has
been associated with various other singular properties of mycobacteria, such as their acid fastness, the granuloma formation, and their
impermeability to antibiotics and solutes [2]. Expectedly, the
sequenced mycobacterial genomes contain a high percent of
genes encoding proteins putatively involved in lipid metabolism.
Mycobacteria, like Gram-negative, contain two membranes
[3]: (1) the plasma membrane whose composition is similar to
other membranes, i.e., a bilayer of phospholipids and proteins,
and (2) an outer membrane (mycomembrane), whose inner leaflet
consists of α-branched and β-hydroxylated C60-C90 fatty acyls
called mycolic acyl (MA) chains, covalently attached to the cell
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_4, © Springer Science+Business Media, LLC, part of Springer Nature 2021
109