Measuring Conjugated Linoleic Acid (CLA) Production by Bifidobacteria

Ahern, Grace; Sinderen, Douwe van; Yang, Bo; Ross, R. Paul; Stanton, Catherine

doi:10.1007/978-1-0716-1274-3_8

Cited by 2 publications

(1 citation statement)

References 43 publications

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“…The crude enzyme (1 mg, quantified by BCA kit) obtained in 2.8 was mixed with 10 mg/mL LA (at a final concentration of 0.3 mg/mL) and 100 μL of 2 mg/mL C15:0 as an internal standard, and left to react at 37 °C with shaking (200 rpm) for 3 h. Once the reaction was complete, the fatty acids were extracted using isopropanol-hexane (v:v 1:2). The absorbance of CLA products was determined as described previously, 33 with a 200 μL hexane layer (containing fatty acids) being removed to a 96-well plate (Greiner Bio-One No. 655801), and the OD value was measured at 233 nm.…”

Section: Identification Of the Bbi-likementioning

confidence: 99%

Computational Analysis and Heterologous Expression of BBI-like Proteins from Food-Grade Bifidobacterium Species Reveal Possibly a Key Factor in Conjugated Linoleic Acid Bioconversion

Mei,

Chen,

Yang

et al. 2023

J. Agric. Food Chem.

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Bifidobacteria are well-known probiotics that can produce multiple health-promoting bioactivities, including the bioconversion of conjugated linoleic acid (CLA). However, there is a lack of insights into the genetic diversity of functional proteins in Bif idobacterium at the species level, particularly owing to the vastly different CLA conversion capabilities of these strains. Here, we performed a comprehensive bioinformatics analysis and the in vitro expression of bbi-like sequences that are widely distributed among CLA-producing Bif idobacterium strains. The BBI-like protein sequences from all four species of bifidobacterial CLAproducing strains were predicted to be stable integral membrane proteins with a transmembrane topology number of seven or nine. All BBI-like proteins were found to be expressed in the Escherichia coli BL21(DE3) hosts with a pure c9, t11-CLA-producing activity. Furthermore, their activities significantly differed in the same genetic background, and their sequence differences were indicated to be important potential factors contributing to the high activity levels in CLA-producing Bifidobacterium breve strains. The use of microorganisms, such as food-grade or industrial-grade strains, to obtain single CLA isomers will accelerate CLA-related food and nutrition research and further enrich the scientific theory of bifidobacteria as probiotics.

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Section: Identification Of the Bbi-likementioning

confidence: 99%