Characterization of the triple-component linoleic acid isomerase in<i>Lactobacillus plantarum</i>ZS2058 by genetic manipulation

Yang, Bo; Qi, Hui; Gu, Zhennan; Zhang, H.; Chen, Wei; Chen, H.; Chen, Y.Q.

doi:10.1111/jam.13570

Cited by 26 publications

(33 citation statements)

References 34 publications

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“…As discussed above, evidence shows the existence of a multienzymatic complex, and studies have confirmed that at least lactobacilli synthesize CFA through a mechanism that may not involve only LAI (84,90,91): the initial step is the hydration of LA by 10-LAH, resulting in 10-HOE acid that is oxidized by a short-chain hydrogenase/oxidoreductase (DH), which is afterwards transformed into 10-oxotrans-11-octadecenoic acid by the action of an acetoacetate decarboxylase (DC) and reduced by DH (10-hydroxy-trans-11-octadecenoic), while finally, LAH leads to the production of RA and C 18:2 t9, t11 (Fig. 3).…”

Section: Multienzymatic Complexes and Possible Synthesis Pathwaysmentioning

confidence: 82%

“…Moreover, in a subsequent study, the same group confirmed, through a gene deletion mutant, that the multiple-component LAI (MCRA, DH, and DC) was responsible for the L. plantarum ZS2058 CLA-producing capacity (91). The mcra gene deletion mutant failed to produce CLA or any intermediate.…”

Section: Cla Production: the Action Of A Multicomponent System?mentioning

confidence: 82%

“…The three genes were then overexpressed in the corresponding deletion mutants, which resulted in each strain recovering the ability to convert LA to CLA. In fact, other species, including Bifidobacterium bifidum, B. breve, Bifidobacterium longum, Lactobacillus casei, L. plantarum, Lactobacillus brevis, Lactobacillus johnsonii, L. reuteri, L. rhamnosus, L. lactis, and Lactococcus cremoris, contained these three genes (91).…”

Section: Cla Production: the Action Of A Multicomponent System?mentioning

confidence: 99%

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Microbial Production of Conjugated Linoleic Acid and Conjugated Linolenic Acid Relies on a Multienzymatic System

Salsinha

Pimentel

Fontes

et al. 2018

Microbiol Mol Biol Rev

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Conjugated linoleic acids (CLAs) and conjugated linolenic acids (CLNAs) have gained significant attention due to their anticarcinogenic and lipid/energy metabolism-modulatory effects. However, their concentration in foodstuffs is insufficient for any therapeutic application to be implemented. From a biotechnological standpoint, microbial production of these conjugated fatty acids (CFAs) has been explored as an alternative, and strains of the genera ,, and have shown promising producing capacities. Current screening research works are generally based on direct analytical determination of production capacity (e.g., trial and error), representing an important bottleneck in these studies. This review aims to summarize the available information regarding identified genes and proteins involved in CLA/CLNA production by these groups of bacteria and, consequently, the possible enzymatic reactions behind such metabolic processes. Linoleate isomerase (LAI) was the first enzyme to be described to be involved in the microbiological transformation of linoleic acids (LAs) and linolenic acids (LNAs) into CFA isomers. Thus, the availability of gene sequences has allowed the development of genetic screening tools. Nevertheless, several studies have reported that LAIs have significant homology with myosin-cross-reactive antigen (MCRA) proteins, which are involved in the synthesis of hydroxy fatty acids, as shown by hydratase activity. Furthermore, it has been suggested that CLA and/or CLNA production results from a stress response performed by the activation of more than one gene in a multiple-step reaction. Studies on CFA biochemical pathways are essential to understand and characterize the metabolic mechanism behind this process, unraveling all the gene products that may be involved. As some of these bacteria have shown modulation of lipid metabolism , further research to be focused on this topic may help us to understand the role of the gut microbiota in human health.

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Section: Multienzymatic Complexes and Possible Synthesis Pathwaysmentioning

confidence: 82%

Section: Cla Production: the Action Of A Multicomponent System?mentioning

confidence: 82%

Section: Cla Production: the Action Of A Multicomponent System?mentioning

confidence: 99%

See 1 more Smart Citation

Microbial Production of Conjugated Linoleic Acid and Conjugated Linolenic Acid Relies on a Multienzymatic System

Salsinha

Pimentel

Fontes

et al. 2018

Microbiol Mol Biol Rev

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“…M206033, China Center for Type Culture Collection) was first cultured in de Man, Rogosa and Sharpe (MRS) medium, which consisted of 1% tryptone, 1% meat extract, 0·5% yeast extract, 2% glucose, 0·1% Tween 80, 0·2% K 2 HPO 4 , 0·5% sodium acetate, 0·2% diammonium citrate, 0·02% MgSO 4 ·7H 2 O and 0·005% MnSO 4 ·H 2 O (pH 6·5) at 37°C in an anaerobic atmosphere (10% H 2 , 10% CO 2 , and 80% N 2 ) in a anaerobic workstation (AW400TG; Electrotek Scientific Ltd, West Yorkshire, UK). Six genetically modified strains of L. plantarum ZS2058 (Yang et al ), including ZS2058Δ mcra , ZS2058Δ mcra /pNZ44‐ mcra , ZS2058Δ dh , ZS2058Δ dh /pNZ44‐ dh , ZS2058Δ dc and ZS2058Δ dc /pNZ44‐ dc, were cultured in MRS medium as the wild type. Escherichia coli BL21 (DE3) carrying the plasmid pET28a harbouring mcra , dh and dc from L. plantarum ZS2058, respectively, were routinely cultured aerobically in Luria–Bertani (LB) medium (1% tryptone, 0·5% yeast extract, 1% NaCl) at 37°C in the presence of kanamycin (10 μg ml −1 ) as a selective marker.…”

Section: Methodsmentioning

confidence: 99%

“…Interestingly, Liavonchanka et al () found that PAI could convert different fatty acids to their conjugated fatty acids, including CLA and CLNA, indicating CLA and CLNA production might share the same enzyme(s) in some bacteria. In our previous study, linoleate isomerase complex for CLA production in L. plantarum ZS2058 was identified via overexpression (Yang et al ) and knockout mutation (Yang et al ). In the current work, the genetic determinates for CLNA generation will be further investigated for L. plantarum ZS2058.…”

Section: Introductionmentioning

confidence: 99%

Genetic determinates for conjugated linolenic acid production in Lactobacillus plantarum ZS2058

Yang

Chen

et al. 2019

J Appl Microbiol

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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).

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Characterization of a recombinant 10‐linoleic acid hydratase from Lactiplantibacillus plantarumZS2058 and biosynthesis of 10‐ hydroxy‐cis‐12‐octadecenoic acid

et al. 2021

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BACKGROUND: 10-Hydroxy-cis-12-octadecenoic acid (10-HOE, 10-OH C18:1), an emerging functional fatty acid, has anti-fungal and anti-inflammatory effects. 10-HOE is synthesized by bacterial 10-linoleic acid hydratase (10-LHT) with linoleic acid as the substate. However, the characterization of 10-LHT and its targeted synthesis of 10-HOE have been rarely reported. In this study, the recombinant 10-LHT from Lactiplantibacillus plantarum ZS2058 was characterized, and the biocatalysis of 10-HOE using crude enzyme was optimized.RESULTS: The recombinant 10-LHT catalyzed the conversion of linoleic acid (C18:2) to 10-HOE as identified using gas chromatography-mass spectrometry (GC-MS). It showed a molecular weight of about 70 kDa on sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), and was a flavin adenine dinucleotide (FAD)-dependent enzyme. The activity of 10-LHT was optimal at pH 6.5 and 25 °C, and it was pH-stable but thermo-sensitive. The optimal condition for the 10-HOE biosynthesis using crude enzyme was 5 g L −1 linoleic acid (C18:2), 148.0 U mL −1 10-LHT, 0.05 mmol L −1 FAD, 2% methanol and 100 mmol L −1 sodium chloride at 25 °C and pH 6.5. A conversion yield of 47.8 ± 1.5% and the corresponding 10-HOE concentration of 2.4 ± 0.1 g L −1 were achieved at 48 h under the optimal reaction conditions. CONCLUSION: This work achieved the highest conversion yield of 10-HOE with the highest substrate concentration, and provides some useful information for the industrial production of 10-HOE.

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

Abstract: Multicomponent linoleic acid isomerase provided important results for illustration unique mechanism for CLA production in Lact. plantarum ZS2058. Lactobacilli with CLA production ability offer novel opportunities for functional food development.

Cited by 26 publications

References 34 publications

Microbial Production of Conjugated Linoleic Acid and Conjugated Linolenic Acid Relies on a Multienzymatic System

Microbial Production of Conjugated Linoleic Acid and Conjugated Linolenic Acid Relies on a Multienzymatic System

Genetic determinates for conjugated linolenic acid production in Lactobacillus plantarum ZS2058

Characterization of a recombinant 10‐linoleic acid hydratase from Lactiplantibacillus plantarumZS2058 and biosynthesis of 10‐ hydroxy‐cis‐12‐octadecenoic acid

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