Genetic determinates for conjugated linolenic acid production in Lactobacillus plantarum ZS2058

Abstract

Aims: To investigate the genetic determinates for conjugated linolenic acid (CLNA) production in Lactobacillus plantarum ZS2058, a high CLNA producer. Methods and Results: After culturing with a-linolenic acid (ALA) in the medium, the fatty acid compositions of supernatant fluid and cell pellets were analysed via GC-MS. cis9,trans11,cis15-CLNA was identified to be the predominant isomer. And during CLNA production, 10-hydroxy-cis12-cis15octadecenoic acid (10-HOEA) and 10-oxo-cis12-cis15-octadecenoic acid (10-OXOA) were accumulated. The E. coli recombinants harbouring genes encoding myosin-cross-reactive antigen (MCRA), short-chain dehydrogenase/ oxidoreductase (DH) and acetoacetate decarboxylase (DC), respectively, were analysed for their roles in CLNA production. The results indicated that MCRA converted ALA to 10-HOEA, following converted to 10-OXOA by DH. While with the combination of three recombinants, ALA could be transformed into CLNA plus 10-HOEA and 10-OXOA. When the three genes were deleted, none of the L. plantarum ZS2058 knockout mutants could produce any CLNA, after complementation, and all the complementary mutants recovered the CLNAproduction ability at similar levels as the wild strain. Conclusions: Lactobacillus plantarum ZS2058 produced CLNA from ALA with 10-HOEA and 10-OXOA as intermediates. The triple-component isomerase of MCRA, DH and DC was the unique genetic determinant for CLNA generation. Significance and Impact of the Study: The current results firstly provided conclusive evidence that the triple-component isomerase complex was shared by both CLA and CLNA production in lactobacilli. ley et al. 2009;Hennessy et al. 2011;Gorissen et al. 2015;Yang et al. 2015). Generally, CLNA could be divided into two parts according to its sources, which are plant-derived CLNA and microbial-derived CLNA. The most studied plant-derived CLNA included jacaranda acid (cis8,trans10, cis12-CLNA), marigold acid (trans8,trans10,cis12-CLNA), pomegranate acid (cis9,trans11,cis13-CLNA) and eleostearic acid (cis9,trans11,trans13-CLNA) (Yuan et al. 2014). Compared with plant-derived CLNA isomers, microbial CLNA showed different conjugated double bonds, such as cis9,trans11,cis15-CLNA, trans9,trans11,cis15-CLNA and cis6,cis9,trans11-CLNA (Hennessy et al. 2012).

c9, t11, c15-CLNA and t9, t11, c15-CLNA from Lactobacillus plantarum ZS2058 Ameliorate Dextran Sodium Sulfate-Induced Colitis in Mice

“…Preparation and Detection of c9, t11, c15-CLNA and t9, t11, c15-CLNA. The α-linolenic acid was isomerized by L. plantarum ZS2058, which can transform approximately 60% of ALA into CLNA, and 31 CLNA1 and CLNA2 Alleviate DSS-Induced Colitis in Mice. The changes in the body weight and disease activity index (DAI) were measured daily during DSS treatment, and the colon length was measured after the mice were euthanized.…”

“…The α-linolenic acid was isomerized by L. plantarum ZS2058, which can transform approximately 60% of ALA into CLNA, and then the CLNA mixtures were extracted from the fermentation. Fatty acids with conjugated double bonds are characterized by an absorption peak at 233 nm.…”

The Arginine Repressor ArgR 2 Controls Conjugated Linoleic Acid Biosynthesis by Activating the cla Operon in Lactiplantibacillus plantarum

“…The enzymes involved in this process included CLA-HY, CLA short-chain dehydrogenase (CLA-DH), and CLA acetoacetate decarboxylase (CLA-DC) ( 16 ). Yang et al also verified this pathway in L. plantarum ZS2058 based on cre-lox gene-editing technology ( 17 , 18 ). Moreover, our previous work had proven that strains with different CLA biosynthesis abilities possessed different transcriptional levels of cla-hy , cla-dh , and cla-dc , suggesting that the upregulation of the CLA yield may be achieved by regulating the transcription of these genes ( 19 ).…”

Antiproliferation Activity and Mechanism of c9, t11, c15-CLNA and t9, t11, c15-CLNA from Lactobacillus plantarum ZS2058 on Colon Cancer Cells

“…Studies on the mechanism of action of this inhibitory activity indicate potential roles for increased cellular lipid peroxidation, altered cellular phospholipid composition, and reductions in the cellular content of Bcl-2 [19]. In our previous work, Lactobacillus plantarum ZS2058 was capable of isomerising α-linolenic acid into CLNA [20]. The structures of Lactobacillus-derived CLNA are c9, t11, c15-CLNA (CLNA1) and t9, t11, c15-CLNA (CLNA2), respectively, where the second or third double bond is in a different position compared with plant-derived CLNA.…”