Synthesis of conjugated linoleic acid by the linoleate isomerase complex in food-derived lactobacilli

Abstract

Aims To assess strains of lactobacilli for their capacity to produce functional fatty acid-conjugated linoleic acid. To assess the linoleate isomerase for CLA production in the most efficient CLA producer. Methods and Results In this study, strains of food-derived lactobacilli were cultured in media with linoleic acid and CLA production was assessed. Most of the selected strains produced CLA at different levels, with Lactobacillus plantarum ZS2058 being the most efficient CLA producer converting over 50% of linoleic acid to c9, t11-CLA and t9, t11-CLA. Some intermediates 10-hydroxy-cis-12-octadecenoic acid, 10-oxo-cis-12-octadecenoic acid and 10-oxo-trans-11-octadecenoic acid were determined via GC-MS. The genes coding the multicomponent linoleate isomerase containing myosin-cross-reactive antigen, short-chain dehydrogenase/oxidoreductase and acetoacetate decarboxylase for CLA production in Lact. plantarum ZS2058 were cloned and expressed in Escherichia coli. With the mixture of recombinant E. coli, c9, t11-CLA and three kinds of intermediates were produced from linoleic acid, which were in line with those in the lactobacilli. Conclusions The ability for CLA production by lactobacilli exhibited variation. Lactobacillus plantarum and Lact. bulgaricus were the most efficient producers in the selected strains. Lact. plantarum ZS2058 converted linoleic acid to CLAs with 10-hydroxy-cis-12-octadecenoic acid, 10-oxo-cis-12-octadecenoic acid and 10-oxo-trans-11-octadecenoic acid as intermediates. The multiple-step reactions for CLA production catalysed by multicomponent linoleate isomerase in Lact. plantarum ZS2058 were confirmed successfully. Significance and Impact of the study Multicomponent linoleate isomerase provides important results for the illustration of the mechanism for CLA production in lactic acid bacteria. Food-derived lactobacilli with CLA production ability offers novel opportunities for functional foods development.

Characterization of the triple-component linoleic acid isomerase inLactobacillus plantarumZS2058 by genetic manipulation

“…According to our previous results (Yang et al . ), three intermediates were produced during CLA production in Lact. plantarum ZS2058.…”

“…; Yang et al . ). Those results indicated that the mechanism for LA isomerization to CLA might be shared by different Lactobacillus species.…”

“…The genetic determinants from Lact. plantarum ZS2058 were cloned in Escherichia coli, respectively, and with the mixture of the recombinants, CLA was produced successfully (Yang et al 2014). The complete genome sequence of Lact.…”

Mining bifidobacteria from the neonatal gastrointestinal tract for conjugated linolenic acid production

“…Fatty acids were derived to corresponding methyl esters with (trimethylsilyl)-diazomethane (Sigma) as described previously. 10,11 The FAMEs were extracted in hexane and loaded on an Rtx-wax column (30 m £ 0.25 mm £ 0.25 mm) using a gas chromatograph (GC2010 plus; Shimadzu, Japan) fitted with a QP2010 Ultra mass spectrometer. Injections of 1 ml were performed automatically at a split ratio of 10:1.…”

“…The temperature program was described previously. 10 The injector and detector were operated at 240 C. Electron energy of 70 eV and ion source temperature of 220 C were used. The percentage conversion to CLNA isomers was calculated by dividing the amount of CLNA present in the broth after inoculation/incubation with the amount of linolenic acid present in the spiked broth before incubation.…”