Draft genome sequence of Lactobacillus delbrueckii subsp. bulgaricus LBP UFSC 2230: a tool for preliminary identification of enzymes involved in CLA metabolism.

Several Lactobacillus ssp. are recognized as potential conjugated linoleic acid (CLA) producers. We have previously reported the ability of a range of Lactobacillus delbrueckii subsp. bulgaricus strains to produce CLA in fermented milk, being a potential candidate for the fermented dairy food chain. This study reports the draft genome sequence of L. bulgaricus strain LBP UFSC 2230, isolated from Italian Grana Padano cheese. Draft genome sequence originated in a total of 4,310,842 paired-end reads that were quality trimmed and assembled into 135 contigs with a total length of 604,745,873 bp, including 2086 protein coding genes and an average GC content of 49.7%. Draft genome sequence represents an important tool to identify the enzymes involved in this strain's CLA metabolism. We identified a gene encoding an enzyme involved in biohydrogenation of linoleic acid pathway, oleate hydratase.

Oleate Hydratase in Lactobacillus delbrueckii subsp. bulgaricus LBP UFSC 2230 Catalyzes the Reversible Conversion between Linoleic Acid and Ricinoleic Acid.

Conjugated linoleic acid (CLA) has been the subject of numerous studies in recent decades because of its associated health benefits. CLA is an intermediate product of the biohydrogenation pathway of linoleic acid (LA) in bacteria. Several bacterial species capable of efficiently converting LA into CLA have been widely reported in the literature, among them Lactobacillus delbrueckii subsp. bulgaricus LBP UFSC 2230. Over the last few years, a multicomponent enzymatic system consisting of three enzymes involved in the biohydrogenation process of LA has been proposed. Sequencing the genome of L. delbrueckii subsp. bulgaricus LBP UFSC 2230 revealed only one gene capable of encoding an oleate hydratase (OleH), unlike the presence of multiple genes typically found in similar strains. This study investigated the biological effect of the OleH enzyme of L. delbrueckii subsp. bulgaricus LBP UFSC 2230 on the hydration of LA and dehydration of ricinoleic acid (RA) and its possible role in the production of CLA. The OleH was cloned, expressed, purified, and characterized. Fatty acid measurements were made by an internal standard method using a gas chromatography-coupled flame ionization detector (GC-FID) system. It was found that the enzyme is a hydratase/dehydratase, leading to a reversible transformation between LA and RA. In addition, the results showed that L. delbrueckii subsp. bulgaricus LBP UFSC 2230 OleH protein plays a role in stress tolerance in Escherichia coli. In conclusion, the OleH of L. delbrueckii subsp. bulgaricus LBP UFSC 2230 catalyzes the initial stage of saturation metabolism of LA, although it has not converted the substrates directly into CLA. IMPORTANCE This study provides insight into the enzymatic mechanism of CLA synthesis in L. delbrueckii subsp. bulgaricus and broadens our understanding of the bioconversion of LA and RA by OleH. The impact of OleH on the production of the c9, t11 CLA isomer and stress tolerance by E. coli has been assisted. The results provide an understanding of the factors which influence OleH activity. L. delbrueckii subsp. bulgaricus LBP UFSC 2230 OleH presented two putative fatty acid-binding sites. Recombinant OleH catalyzed both LA hydration and RA dehydration. OleH was shown to play a role in bacterial growth performance in the presence of LA.

Biohydrogenation of Linoleic Acid by Lactic Acid Bacteria for the Production of Functional Cultured Dairy Products: A Review.

Conjugated linoleic acid (CLA) isomers have attracted significant attention due to their important physiological properties, which have been observed in humans. Many lactic acid bacteria (LAB) demonstrate the ability to produce CLA isomers (C18:2 cis-9, trans-11 and C18:2 trans-10, cis-12) from the linoleic acid (LA) present in milk or in synthetic media. CLA isomers can be synthesized in vitro by LAB using vegetable oils rich in LA. The aim of this review is to present an update on the studies that have been conducted on the production of CLA isomers from LA mainly by LAB and of the factors that influence this conversion (source and concentration of LA and fermentation conditions). In addition, this review presents the relationship between the consumption of CLA isomers and their health benefits in humans such as anti-atherosclerosis and anti-carcinogenic effects. There is considerable variation between the studies concerning the beneficial effects of CLA in animal models, which have not been reflected in human studies. This can be attributed to the differences in the doses of CLA isomers used and to the different sources of CLA. Furthermore, the regulatory and scientific information classifying the physiological properties of CLA, which serve as support for the claims of its potential as a functional ingredient, are presented. More research is needed to determine whether CLA production by LAB can be enhanced and to determine the optimal requirements for these microbial cultures. Furthermore, safety and efficacy of CLA consumption have to be investigated in the future.