الفهرس 
: Production of Bioactive Compounds as Anticancer from Probiotics
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Abstract
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Probiotics have shown promising results in prophylaxis and safe biotherapy. The
search for a biologically active strain able to produce bioactive compounds of applied
importance to meet the growing needs was the main target. To attain this target, buffalo
colostrum samples was used as an isolation source of LAB. Out of the 53 LAB isolates,
only 16 produced exopolysaccharides; of which, one isolate attained the highest
productivity (3.8 g l-1) and was also able to produce L-asparaginase (ASNase). Such isolate
was molecularly identified as Weissella paramesenteroides MN2C2 and deposited in the
GenBank under number MK530206. The optimum conditions for maximal productivity of
bioactive compounds were 24 h incubation at 35 °C and pH 6.5 for EPSs; 48 h at 37 °C,
pH 7 and 10% inoculum concentration for ASNase. EPSs molecules were partially purified
(PP) and physicochemical characterized; UV, FTIR spectra showed absorption bands
related to many functional groups. 1HNMR polysaccharide spectrum consists mainly of
three regions: ring proton of several sugar residues of the polysaccharides, anomeric proton
region and the alkyl proton region. HPLC elucidated EPSs structure as consisting of 80%
fructose, 9.3% glucose and 5.6% sucrose. SEM showed a three-dimensional structure of
irregular highly compacted lumps with different sizes and a smooth surface. Energy
dispersive X-Ray and mapping analyses revealed that PP-EPSs composed of carbon,
nitrogen, oxygen, phosphorous and sulfur in high ratios of its weight reached 42.31, 10.11,
42.68, 4.13 and 0.78 %, respectively. As indicated by TEM and DLS analyses, the
nanoparticle (NP) sizes of EPSs prepared from crude or PP using ultrasonication ranged
from 37.3 to 105 nm for the crude CEPSs-NPs, and from 45.7 to 204 nm for the PPEPSs-
NPs, PP-ASNase has a specialized activity of 72 units mg-1 protein and a molecular weight
of ≈ 36 kDa using SDS-PAGE technique. PP-EPSs proved its superiority as anticancer with
highly selectivity indices against liver HepG-2, colon (Caco-2) and breast MCF-7 cancer
cells. The CEPSs-NPs had significant activity against breast cancer MCF-7; and the toxicity
was increased in the nanoparticles prepared from partially purified polysaccharides. Both
CEPS-NPs and PPEPSs-NPs also showed stronger DPPH antioxidant activity than CEPSs,
PP-EPSs and PP-ASNase, respectively. Both PP-EPSs and CEPSs reduced Coxsackievirus
(CVB3) yield by > 99%. The probiotic W. paramesenteroides MN2C2 were characterized
by its survivability for 3 and 6 hours at pH ranging from 1.5 to 9 and 0.3% of bile salts; in
the presence of pancreatic juice, under simulated stomach and intestinal conditions; besides
its ability to adhere to intestinal walls. It also showed antimicrobial activity against B.
subtilis and P. aeruginosa, with inhibition zones of 25 and 19 mm, respectively. The strain
is considered as safe because of the absence of analytical activity in human blood and the
high sensitivity to antibiotics tetracycline, fusidic acid, ampicillin and resistance to
vancomycin and kanamycin. Also, W. paramesenteroides MN2C2 was able to ferment milk
when mixed with probiotic commercial starter strains to produce a promising milk product
that is suitable, practical, effective, preventative and with safe therapeutic properties