Article
J. Agric. Food Chem., Vol. 57, No. 17, 2009 7705
Recently (26), food and human intestinal bacterial strains were
screened for cinnamoyl esterase activity. The enzymatic hydro-
lysis of RA results in the generation of CA and DHPL. CA has
antioxidant properties, illustrated by its ability to scavenge
various free radicals when tested in vitro (27, 28). In vivo, when
ingested with the diet, CA increases the plasma antioxidant
capacity, the concentration of endogenous antioxidants such as
vitamin E, and the ex vivo resistance of lipoproteins to oxida-
tion (29-31).
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DHPL is one of the most widely used traditional Chinese
medicines for the treatment of various cardiovascular diseases
and has been reported to have potential protective effects from
oxidative injury (32, 33). DHPL has also been used for its radical
scavenging and antioxidant activities as well as for the treatment
of cerebrovascular diseases (34). In the present investigation, RA
was enzymatically hydrolyzed into CA and DHPL. Among tested
enzymes, only chlorogenate esterase (A. japonicus) was able to
perform this hydrolysis. This result confirms the hypothesis that
RA could be cleaved by a cinnamoyl esterase. The enzymatic
hydrolysis of RA to produce CA and DHPL could be an
interesting approach for applications in food products. Cinna-
moyl esterases have been commonly found in rumen and soil
saprophytic microorganisms (35) and in bacteria from human
and animal intestinal microbiota (24). L. johnsonii NCC 533 (La1)
used in this study is a probiotic strain isolated from the human
intestinal microbiota. Complete hydrolysis of RA was achieved
with La1 cells, while no cinnamoyl esterase-like activity was
identified in both culture and reaction media. Incubation of RA
with La1 crude extract resulted in a strong increase of the reaction
rate as compared to nonlysed cells. These observations suggest
that the enzyme involved in the hydrolysis of RA is presumably
intracellular. The identification, purification, and characteriza-
tion of La1 esterase involved in RA hydrolysis are under
investigation. Treatment of rosemary extract with La1 cells
showed a complete degradation of RA, while the other key
molecules remained intact as shown by LC-MS. The La1 was
selected to perform the hydrolysis of RA because this bacterium is
known to possess several esterases (genome data). Other lactic
acid bacteria were screened and used for their cinnamoyl esterase
activity (26).
Experiments in the GI tract model (TIM-1) were conducted to
see whether the hydrolysis of RA occurs chemically under the
conditions of the GI model (e.g., temperature, pH, and bile salts),
catalyzed by secreted enzymatic activity (e.g., lipase and pancrea-
tic enzymes), or by selected enzymes and microorganisms.
No hydrolysis of RA was observed under the physiological
conditions mimicked in the TIM-1 model. The hydrolysis of
RA was not catalyzed by secreted enzymatic activity. However,
substantial hydrolysis of RA was observed when La1 was added
to rosemary extract in the TIM-1 model. These results confirm
the hypothesis that RA and probably hydroxycinnamates are
degraded by gut microflora before their absorption and are then
metabolized in various tissues such as intestine, liver, and kidney.
In conclusion, this is the first time that cinnamoyl esterase-like
activity is identified in the probiotic L. johnsonii (La1). The results
obtained in this study support the hypothesis that RA is degraded
by gut microflora before absorption and metabolization.
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Received April 30, 2009. Revised manuscript received July 15, 2009.
Accepted July 23, 2009.
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