Bioactive Compounds and In Vitro Antioxidant and Anticoccidial Activities of Opuntia ficus-indica Flower Extracts.

The objective of the present study is to identify the biochemical compounds extracted from OFI flowers using ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry and to evaluate their in vitro antioxidant activities and anticoccidial effects on the destruction of Eimeria oocysts isolated from naturally infected chickens. A domestic microwave was used with a refrigerant to condense the vapors generated during the extraction. The flavonoid and phenolic compound contents of the OFI flowers were determined according to standard methods. DPPH radical and H2O2 scavenging capacities were used to assess the antioxidant activity. Regarding the anticoccidial activity, the Eimeria spp. oocysts used were isolated from the fresh feces of infected broilers and were determined in triplicate by incubation at an ambient temperature for 24 h. The results highlighted the considerable influence of the optimized acetone concentration, ratio, irradiation time, and microwave power parameters on the phenolic content and antioxidant activities. Our results revealed significant matches between the predicted and experimental values of the models. Molecular analysis revealed the presence of several biophenol classes such as quercetin, isorhamnetin 3-O-rutinoside, and quercetin-3-O-rutinoside. OFI flower extracts inhibited sporulation and damaged the morphology of Eimeria oocysts compared with normal sporulated Eimeria oocysts containing sporocysts. In conclusion, the optimized conditions were validated and found to fit very well with the experimental values. These findings suggest that the flowers of OFI should be considered sources of antioxidants. The results of the present study revealed that OFI flower extracts have anticoccidial activities against Eimeria-spp.-induced infection in broiler chickens.

Statistical Optimization and Partial Characterization of Xylanases Produced by Streptomyces sp. S1M3I Using Olive Pomace as a Fermentation Substrate.

Xylanase production by Streptomyces sp. S1M3I was optimized by response surface methodology (RSM), followed by a partial characterization of these enzymes. Olive pomace was used as a substrate for growing Streptomyces sp. S1M3I in submerged fermentation. Effects of incubation time, pH, temperature, carbon source, nitrogen source, and inoculum size on xylanase production were studied, through the one-factor-at-a-time method. Then, a 33-factorial experimental design with RSM and the Box-Behnken design was investigated for the major influence factors. Maximum xylanase production (11.28 U/mL) was obtained when the strain was grown in mineral medium supplemented with 3% (w/v) of olive pomace powder and 0.3% (w/v) of ammonium sulfate, at a pH 7.4 and an incubation temperature of 40 °C. The xylanases in the supernatant degraded all tested substrates, with higher activity for the low-viscosity wheat arabinoxylan substrate. Two xylanases with close molecular masses were detected by zymogram analysis: Xyl-1 and Xyl-2 with molecular masses of 24.14 kDa and 27 kDa, respectively. The optimization of enzyme production parameters of Streptomyces sp. S1M3I and the characterization of these enzymes are prerequisites to enhancing xylanase production yield, which is crucial for further biotechnological processes.

Peroxidase-producing actinobacteria from Algerian environments and insights from the genome sequence of peroxidase-producing Streptomyces sp. S19 / Actinobacteria productora de peroxidasa de ambientes argelinos y perspectivas de la secuencia genómica de Streptomyces sp. S19

Unique environments often serve as a source of novel microorganisms with novel chemistries. In this study, telluric samples collected from different regions of Algeria were processed for the isolation of novel peroxidase-producing actinobacterial strains. An agar-based screening identified 45 isolates with the ability to produce peroxidase. The 16S rRNA gene sequencing showed that most of the strains belong to the genus Streptomyces. Optimization of cultivation conditions was performed for the top five peroxidase-producing strains. Apart from strain 36 (optimal growth temperature of 30 °C) and strain 45 (optimal medium pH of 6.0), the strains exhibited optimal peroxidase production when cultivated for 5 days at 37 °C and in a medium at pH 7.0. Extracellular peroxidase production was induced by ferulic acid in three of the five strains, while the presence of canola lignocellulosic waste (CLW) induced peroxidase production in all strains. Strain 19 (S19) was selected for further optimization and the extracellular peroxidase purified using acid and acetone precipitation, followed by size exclusion chromatography. The purified fraction showed a single band on the polyacrylamide gel with an estimated molecular weight of 21.45 kDa. Genome analysis confirmed the assignment of S19 to the genus Streptomyces, the presence of genes encoding for peroxidases, and the presence of genes encoding for carbohydrate-active enzymes. The presence of biosynthetic gene clusters potentially encoding for biosurfactants further highlighted the great biotechnological potential of the strain.(AU)

Peroxidase-producing actinobacteria from Algerian environments and insights from the genome sequence of peroxidase-producing Streptomyces sp. S19.

Unique environments often serve as a source of novel microorganisms with novel chemistries. In this study, telluric samples collected from different regions of Algeria were processed for the isolation of novel peroxidase-producing actinobacterial strains. An agar-based screening identified 45 isolates with the ability to produce peroxidase. The 16S rRNA gene sequencing showed that most of the strains belong to the genus Streptomyces. Optimization of cultivation conditions was performed for the top five peroxidase-producing strains. Apart from strain 36 (optimal growth temperature of 30 °C) and strain 45 (optimal medium pH of 6.0), the strains exhibited optimal peroxidase production when cultivated for 5 days at 37 °C and in a medium at pH 7.0. Extracellular peroxidase production was induced by ferulic acid in three of the five strains, while the presence of canola lignocellulosic waste (CLW) induced peroxidase production in all strains. Strain 19 (S19) was selected for further optimization and the extracellular peroxidase purified using acid and acetone precipitation, followed by size exclusion chromatography. The purified fraction showed a single band on the polyacrylamide gel with an estimated molecular weight of 21.45 kDa. Genome analysis confirmed the assignment of S19 to the genus Streptomyces, the presence of genes encoding for peroxidases, and the presence of genes encoding for carbohydrate-active enzymes. The presence of biosynthetic gene clusters potentially encoding for biosurfactants further highlighted the great biotechnological potential of the strain.

Differential antioxidant activity of glucuronoxylooligosaccharides (UXOS) and arabinoxylooligosaccharides (AXOS) produced by two novel xylanases.

XOS are particularly interesting bioactive molecules. Bacillus safensis CBLMA18, a xylanolytic bacterium has been isolated and two of its xylanases have been identified and fully characterized. Xyn11A is an extracellular 22.5-kDa GH11 xylanase while a second xylanase, Xyn10B, corresponds to an intracellular 48-kDa GH10 enzyme. Both unimodular xylanases showed activity only on xylan substrates with important differences in their catalytic pattern. Xyn11A displays higher activity on glucuronoxylans, with an optimum at pH 8 and 50 °C, and a Vmax of 5281 U/mg on beechwood xylan, meanwhile Xyn10B prefers arabinoxylans, with an optimum of pH 7 and 60 °C, and a Vmax of 50.29 U/mg on rye arabinoxylan. The antioxidant activity of xylanase-generated XOS obtained from glucuronoxylans (UXOS) and arabinoxylans (AXOS) was tested with the ABTS (2, 2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) method. UXOS showed higher antioxidant activity than AXOS (>80% of antioxidant capacity). Thin layer chromatography and MALDI-TOF MS analysis showed that UXOS comprise neutral and acidic XOS with methylglucuronic acid (MeGlcA) ramifications, while AXOS contain only neutral molecules with arabinose decorations. The MeGlcA ramifications seem to have an important role in the antioxidant capacity of oligosaccharides. Besides, the increase of UXOS size correlates with an increase in their activity.

Biochemical properties of a new thermo- and solvent-stable xylanase recovered using three phase partitioning from the extract of Bacillus oceanisediminis strain SJ3.

The present study investigates the production and partial biochemical characterization of an extracellular thermostable xylanase from the Bacillus oceanisediminis strain SJ3 newly recovered from Algerian soil using three phase partitioning (TPP). The maximum xylanase activity recorded after 2 days of incubation at 37 °C was 20.24 U/ml in the presence of oat spelt xylan. The results indicated that the enzyme recovered in the middle phase of TPP system using the optimum parameters were determined as 50% ammonium sulfate saturation with 1.0:1.5 ratio of crude extract: t-butanol at pH and temperature of 8.0 and 10 °C, respectively. The xylanase was recovered with 3.48 purification fold and 107% activity recovery. The enzyme was optimally active at pH 7.0 and was stable over a broad pH range of 5.0-10. The optimum temperature for xylanase activity was 55 °C and the half-life time at this temperature was of 6 h. At this time point the enzyme retained 50% of its activity after incubation for 2 h at 95 °C. The crude enzyme resist to sodium dodecyl sulfate and ß-mercaptoethanol, while all the tested ions do not affect the activity of the enzyme. The recovered enzyme is, at least, stable in tested organic solvents except in propanol where a reduction of 46.5% was observed. Further, the stability of the xylanase was higher in hydrophobic solvents where a maximum stability was observed with cyclohexane. These properties make this enzyme to be highly thermostable and may be suggested as a potential candidate for application in some industrial processes. To the best of our knowledge, this is the first report of xylanase activity and recoverey using three phase partitioning from B. oceanisediminis.