Production of Conjugated Linoleic Acid by Lactic Acid Bacteria: Screening and Optimization

Angelo M. Tapia1*, Jose Arceo N. Bautista1, Bernadette C. Mendoza2,
Laura J. Pham3, Ione G. Sarmago4, and Maria Cynthia R. Oliveros1

1Institute of Animal Science, College of Agriculture and Food Science
2Institute of Biological Sciences, College of Arts and Sciences
3National Institute of Molecular Biology and Biotechnology
4Dairy Training and Research Institute, College of Agriculture and Food Science
University of the Philippines Los Baños (UPLB), Laguna 4030 Philippines

*Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.




This study was conducted to screen and optimize locally isolated lactic acid bacteria (LABs) for conjugated linoleic acid (CLA) production. Ten (10) LAB strains were evaluated for CLA production in De Man, Rogosa, and Sharpe (MRS) broth supplemented with 0.01% free linoleic acid (LA). Lipids were extracted from the broth with chloroform/methanol. The resultant fatty acid methyl esters were further extracted with n-hexane and were analyzed by gas chromatography using a Shimadzu GC-14 unit (Shimadzu Corp., Kyoto Japan) equipped with SUPELCOWAX 10 column. Three (3) LAB strains – Lactobacillus plantarum 1066, Lactobacillus fermentum 1014, and Lactobacillus casei 1064 – were able to produce CLA. L. plantarum 1066 was selected for optimization where LA supplementation (100 and 200 µg/ml) and incubation time (0, 6, 12, 18, and 24 h) were used as variable factors for CLA production. Supplementation with LA at 100 or 200 µg/ml in MRS broth did not significantly affect the growth of L. plantarum 1066 after 24 h of incubation, as well as its production of CLA. However, it was observed that CLA production showed a strong positive correlation with LA concentration.



CLAs are isomers of the polyunsaturated fatty acid LA. It is a mixture of positional and geometric isomers of LA (c-9, c-12 C-18:2-LA) with two conjugated unsaturated double bonds at various carbon positions (c-9, c-11 and c-10, c-12). It is formed as an intermediate during the biohydrogenation of LA to stearic acid by Butyrivibrio fibrisolvens (Kepler and Tove 1967) and other rumen bacteria (Kritchevsky 2000) or from the endogenous conversion of transvaccenic acid (t-11 C, 18:1 TVA) by 9-desaturase in the mammary gland (Corl et al. 2001). . . . . read more



ALCALA LM, FONTECHA J. 2007. Fatty acid and conjugated linoleic acid (CLA) isomer composition of commercial CLA-fortified dairy products: evaluation after processing and storage. J Dairy Sci 90(5): 2083–2090.
ALONSO L, CUESTA EP, GILLILAND SE. 2003. Conjugated linoleic acid by Lactobacillus acidophilus and Lactobacillus casei of human intestinal origin. J Dairy Sci 86: 1941–1946.
ANDRADE JC, ASCENCAO K, GULLON P, HENRIQUEZ S, PINTO J, ROCHA SANTOS T. 2012. Production of CLA by food-grade bacteria – A review. Int Dairy J 65: 467–481.
BELURY MA. 2002. Inhibition of carcinogenesis by conjugated linoleic acid: Potential mechanisms of action. Recent Advances in Nutritional Sciences 132(10): 2995–2998.
BLIGH EG, DYER JW. 1959. A rapid method of total lipid extraction and purification. Can J Biochem Phys 37(8): 911–917.
COAKLEY M, ROSS RP, NORDGREN M, FITZGERALD G, DEVERY R, STANTON C. 2003. Conjugated linoleic acid biosynthesis by human-derived Bifidobacterium sp. J Appl Microbiol 94(1): 138–145.
COLLOMB M, SCHMID A, SIEBER R, WECHSLER D, RYHANEN E. 2006.  Conjugated linoleic acid in milk fat: Variation and physiological effects. Int Dairy J 16(11): 1347–1361.
COOK ME, PARIZA MW. 1998. The role of conjugated linoleic acid (CLA) in health. Int Dairy J 8(5): 459–462.
CORL BA, BAUMGARD LH, DWYER DA, GRIINARI JM, PHILLIPS BS, BAUMAN DE. 2001. The role of delta (9)-desaturase in the production of cis-9, trans-11 CLA. J Nutr Biochem 12(11): 622–630.
CSAPO J, VARGA V. 2015. Advances in fermented foods and beverages, 1st ed. Cambridge, UK: Elsevier Ltd. p. 78.
GREENWAY DL, DYKE KG. 1979. Mechanism of the inhibitory action of linoleic acid on the growth of Staphylococcus aureus. J Gen Microbiol 115(1): 233–245.
GORRISEN L, DE VUYST L, RAES K, DE SMET S. 2016. Conjugated linoleic and linoleic acid production kinetics by bifidobacteria differ among strains. Int J Food Microbiol 155(1): 234–240.
GUROVIC M, ENTILLI A, OLIVERA N, RODRIGUEZ M. 2014. Lactic acid bacteria isolated from fish gut produce conjugated linoleic acid without the addition of exogenous substrate. Process Biochem 49(7): 1071–1077.
HEIPIEPER HJ, MEINHARDT F, SEGURA A. 2003. The cis-trans isomerase of unsaturated fatty acids in Pseudomonas and Vibrio: Biochemistry, molecular biology and physiological function of a unique stress adaptive mechanism. FEMS Microbiol Lett 229(1): 1–7.
JIANG J, BJORK L, FONDLEN R. 1998. Production of conjugated linoleic acid by dairy starter cultures. J Appl Microbiol 86(1): 95–102.
KEPLER CR, TOVE SB. 1967. Biohydrogenation of unsaturated fatty acids. J Biol Chem 242(1): 5686–5692.
KIM YJ, LIU RH. 2002. Increase of conjugated linoleic acid content in milk by fermentation with lactic acid bacteria. J Food Sci 67(5): 1731–1737.
KISHINO S, OGAWA J, OMURA Y, MATSUMURA K, SHIMIZU S. 2002. Conjugated linoleic acid production by lactic acid bacteria. Journal of the American Oil Chemists’ Society 79(2): 159–163.
KRITCHEVSKY D. 2000. Conjugated linoleic acid. Nutrition Bulletin 25: 25–28.
LAWSON RE, MOSS AR, GIVEN DI. 2001. The role of dairy products in supplying conjugated linoleic acid to man’s diet. Nutrition Research Reviews 14(1): 153–172.
LIN TY. 2003. Influence of lactic cultures, linoleic acid and fructo-oligosaccharides on conjugated linoleic acid concentration in non-fat set yogurt. Aust J Dairy Technol 58(2): 11–14.
MCGUIRE MA, MCGUIRE MK. 2000. Conjugated linoleic acid (CLA): A ruminant fatty acid with beneficial effects on human health. J Anim Sci 77: 1–8.
MOLONEY F, YEOW T, MULLEN A, MOLAN JJ, ROCHE HM. 2004. Conjugated linoleic acid supplementation, insulin sensitivity, and lipoprotein metabolism in patients with type 2 diabetes mellitus. Am J Clin Nutr 80(4): 887–895.
NIEUWENHOVE CP, LISZEWSKI R, GONZALEZ SN, PEREZ AB. 2007. Conjugated linoleic acid conversion by dairy bacteria cultured in MRS broth and buffalo milk. Lett Appl Microbiol 44(5): 467–474.
OGAWA J, KISHINO S, ANDO A, SUGIMOTO S, MIHARA K, SHIMIZU S. 2005. Production of conjugated linoleic acid by lactic acid bacteria. J Biosci Bioeng 100(4): 355–364.
PARIZA MW. 2004. Perspective on the safety and effectiveness of conjugated linoleic acid. Am J Clin Nutr 79(6): 1132–1136.
PHAM LJ, GREGORIO MB. 2008. The triacylglycerol structure of coconut oil determined by chromatography combined with stereospecific analysis. Philipp Agri Sci 91(3): 343–347.
SIRUANA A, CALSAMIGLIA S. 2016. A meta-analysis of feeding strategies to increase the content of conjugated linoleic acid (CLA) in dairy cattle milk and the impact on daily human consumption. Anim Feed Sci Tech 217: 13–26.
TERAN V, LUNA-PIZARRO D, ZACARIAS M, VINDERTA G, MEDINA R, VAN NIEWENHOVE C. 2015. Production of conjugated dienoic and trienoic fatty acids by lactic acid bacteria and bifidobacteria. J Funct Foods 19: 417–425.