Heterologous expression of linoleic acid isomerase from Propionibacterium acnes and anti-proliferative activity of recombinant trans-10, cis-12 conjugated linoleic acid.
The linoleic acid isomerase enzyme from Propionibacterium acnes responsible for bioconversion of linoleic acid to trans-10, cis-12 conjugated linoleic acid (t10, c12 CLA) was cloned and overexpressed in Lactococcus lactis and Escherichia coli, resulting in between 30 and 50 % conversion rates of the substrate linoleic acid to t10, c12 CLA. The anti-proliferative activities of the fatty acids produced following isomerization of linoleic acid by L. lactis and E. coli were assessed using the human SW480 colon cancer cell line. Fatty acids generated from both L. lactis and E. coli contained a mixture of linoleic acid and t10, c12 CLA at a ratio of approximately 1.35 : 1. Following 5 days of incubation of SW480 cells with 5-20 microg ml(-1) (17.8-71.3 microM) of the t10, c12 CLA, there was a significant (P<0.001) reduction in growth of the SW480 cancer cells compared with the linoleic acid control. Cell viability after treatment with the highest concentration (20 microg ml(-1)) of the t10, c12 CLA was reduced to 7.9 % (L. lactis CLA) and 19.6 % (E. coli CLA), compared with 95.4 % (control linoleic acid) and 31.7 % (pure t10, c12 CLA). In conclusion, this is believed to represent the first report in which recombinant strains are capable of producing CLA with an anti-proliferative potential.
Microbiology Department, University College Cork, Ireland.|Teagasc, Moorepark Food Research, Biotechnology Centre, Fermoy, Co. Cork, Ireland.
Eva
E
Rosberg-Cody
Alimentary Pharmabiotic Centre, Cork, Ireland.|Teagasc, Moorepark Food Research, Biotechnology Centre, Fermoy, Co. Cork, Ireland.
Mark C
MC
Johnson
Alimentary Pharmabiotic Centre, Cork, Ireland.|Microbiology Department, University College Cork, Ireland.
Gerald F
GF
Fitzgerald
Alimentary Pharmabiotic Centre, Cork, Ireland.|Teagasc, Moorepark Food Research, Biotechnology Centre, Fermoy, Co. Cork, Ireland.
Paul R
PR
Ross
Alimentary Pharmabiotic Centre, Cork, Ireland.|Teagasc, Moorepark Food Research, Biotechnology Centre, Fermoy, Co. Cork, Ireland.
Catherine
C
Stanton
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001711:Biotransformation; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006131:Growth Inhibitors / Q000494:pharmacology; D006801:Humans; D007535:Isomerases / Q000235:genetics* / Q000378:metabolism*; D013294:Lactococcus lactis / Q000235:genetics; D019787:Linoleic Acid / Q000378:metabolism*; D044243:Linoleic Acids, Conjugated / Q000494:pharmacology*; D011425:Propionibacterium acnes / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D000970:Antineoplastic Agents; D006131:Growth Inhibitors; D044243:Linoleic Acids, Conjugated; D011994:Recombinant Proteins; C496197:trans-10,cis-12-conjugated linoleic acid; D019787:Linoleic Acid; D007535:Isomerases