Lactobacillic acid

Lactobacillic acid, scientifically 10-(2-hexylcyclopropyl)­ decanoic acid, is a naturally occurring chemical compound from the group of fatty acids. Its salts are called lactobacillates. A special feature is the cyclopropane ring in the carbon chain. Lactobacillic acid, with 19 carbon atoms, is an odd-chain fatty acid. The fatty acid was detected in the 1950s in bacteria of the genus Lactobacillus, but is also found in numerous other bacterial species. The bacterial biosynthesis of lactobacillic acid takes place from cis-vaccenic acid (cis-11-octadecenoic acid), a unsaturated fatty acid that has one carbon atom less. Bacteria in a batch culture form the fatty acid at the end of the exponential phase of growth or in the early stationary growth phase. Previous studies have shown that the biosynthesis and storage of lactobacillic acid in the cell membrane is associated with a protective effect for the bacterial cells, although the exact mechanism has not been conclusively clarified. In bacteriology, the fatty acid is mainly used for analytical purposes, for example in the identification of bacteria.

History

Discovery In the 1950s, a working group at the University of Pittsburgh conducted research on bacteria of the genus Lactobacillus, which require biotin as a growth factor. It has previously been shown that biotin is no longer necessary for the growth of bacteria if certain fatty acids are present in the culture medium instead. When investigating the influence of biotin on fatty acid metabolism, the researchers initially concentrated on the species L. arabinosus, which according to current systematics is classified as L. plantarum is carried out. They cultivated the bacteria in a semi-synthetic culture medium, harvested the cells and extracted the "free" lipids with acetone and diethyl ether. This fraction makes up about 20% of the total lipids. To obtain the "bound" lipids, an acid hydrolysis was then carried out, in which fatty acids bound as esters were released and also extracted with diethyl ether. The fatty acids were methylated with diazomethane to the methyl esters and separated according to their boiling points using fractional distillation. Based on the distillation curve, the presence of esters of C16, C18 and C19 fatty acids was expected. The fatty acid obtained from the C19 fraction showed a melting point at 28-29 °C after purification by recrystallization. The compound was investigated using numerous physical and chemical methods and its molecular formula was determined as C19H36O2. In 1950, this was only the second fatty acid with 19 carbon atoms to be isolated from microorganisms. As early as 1929, the tuberculostearic acid, molecular formula C19H38O2) was discovered, which was isolated from the tuberculosis pathogen Mycobacterium tuberculosis. In analogy to its name, the biochemists suggested the English name 'lactobacillic acid', as the fatty acid originated from a Lactobacillus species. Lactobacillic acid has also been isolated from L. casei. In the publication, the scientists emphasized the implications of their discovery, which contradicted the opinion at the time that only fatty acids with an even number of carbon atoms occur in nature.

Structure elucidation

The "free" and "bound" lipids do not differ significantly in their fatty acid composition. In addition to lactobacillic acid with a share of 31%, palmitic acid (C16:0), stearic acid (C18:0) and cis-vaccenic acid (C18:1 cis-11) with a proportion of 37%, 2% and 20% respectively. The test results of the newly discovered fatty acid showed that it is a saturated fatty acid. It is stable towards oxidizing agents that would react with a double bond in the carbon chain. In the reaction with hydrogen bromide (HBr), however, an addition of HBr occurs in the molecule. Hydrogenation is also possible, resulting in several isomers fatty acids with the molecular formula C19H38O2, one of which has been identified as nonadecanoic acid. The other compound is a branched-chain fatty acid with a methyl group as a branch (methyloctadecanoic acid), although the scientists could not distinguish at the time whether one or more isomers of it were present. Based on the results of the chemical and physical (infrared spectroscopy and X-ray diffraction) methods for structure elucidation, a saturated fatty acid with a cyclopropane ring in the carbon chain was proposed as the structure.

from its lipids. By comparison with synthetically produced fatty acids, they were able to determine the position of the cyclopropane ring and proposed the name 11,12-methylene­octadecanoic acid, without specifying the stereoisomerism of the structure. Viewed from the cyclopropane ring, different substituents are present on two carbon atoms, resulting in cis-trans-isomerism (see figure). The substituents can be on the same side (cis) or on different sides (trans) of the ring bond. Hofmann et al. were initially unable to clarify which structure was actually present at the time of discovery. However, in 1954 they hypothesized that the cis isomer was present. This was confirmed by a Canadian research group in 2005. A clear description of the spatial arrangement of the substituents is possible with the help of the Cahn-Ingold-Prelog priority rules, according to which the absolute configuration of the molecule is given as 11R,12S. The derived name (11R,12S)-methylenoctadecanoic acid is commonly used, even if it does not correspond to the recommendations of the IUPAC nomenclature. For the systematic naming of lactobacillic acid, the absolute configuration is given from the cyclopropane ring, with a C6 substituent (hexyl group) and a C10 substituent containing the carboxy group (decanoic acid), thus giving 10-[(1R,2S)-2-hexyl­cyclopropyl]­decanoic acid

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