Functional and structural characterization of family 6 carbohydrate-binding module (CtCBM6A) of Clostridium thermocellum α-L-Arabinofuranosidase.

The gene encoding the family 6 carbohydrate-binding module (CtCBM6A) from Clostridium thermocellum, cloned in pET-21a(+) expression vector, was overexpressed using Escherichia coli BL-21(DE3) cells and purified by immobilized metal-ion affinity chromatography. SDS-PAGE analysis of the recombinant CtCBM6A showed molecular size of approximately 15 kDa. Ligand-binding analysis of CtCBM6A with rye arabinoxylan and oat spelt xylan by affinity gel electrophoresis showed low affinity for these ligands (Ka of 40 and 26 liter/g, respectively), and analysis by fluorescence spectroscopy (Ka of 33 and 15 liter/g, respectively) corroborated lower binding affinity with the above soluble ligands. However, CtCBM6A displayed significantly higher ligand-binding affinity with insoluble wheat arabinoxylan with equilibrium association constant Ka of 230 M(-1) and binding capacity (N0) of 11 µmole/g. The protein melting curve of CtCBM6A displayed a peak shift from 53 to 58°C in the presence of Ca2+, indicating that Ca2+ imparts thermal stability to the CtCBM6A structure. Homology modeling of CtCBM6A revealed a characteristic ß-sandwich core structure. The Ramachandran plot of CtCBM6A showed 89% of the residues in the most favorable region, 10% in additionally favored region, and 1% in generously allowed region, indicating that CtCBM6A has a stable conformation.

Pasture intake improves the performance and meat sensory attributes of free-range broilers.

Free-range chickens are assumed to consume low to moderate levels of pasture, although the effects of forage intake in broiler performance and poultry meat quality remain to be established. In addition, despite cellulases and hemicellulases being widely used as feed supplements to improve the nutritive value of cereal-based diets for fast-growing broilers, the potential interest of these biocatalysts in the production of free-range chicken is yet to be established. In this study, broilers of the RedBro Cou Nu x RedBro M genotype were fed a cereal-based diet in portable floorless pens located either on a rainfed subterranean clover (Trifolium subterraneum) pasture or on an irrigated white clover (Trifolium repens) pasture. Control birds were maintained at the same site in identical pens but with no access to pastures. The importance of pasture intake and enzyme supplementation in the performance and meat sensory properties of the free-range chicken from d 28 to 56 was investigated. The results revealed that although cellulase and hemicellulase supplementation had no impact on broiler performance (P > 0.05), birds foraging on legume-based pastures reached significantly greater final BW. The data suggest that the improvement in broiler performance results from increased intake of the cereal-based feed rather than from an improvement in the efficiency of nutrient utilization per se. Interestingly, although the intake of the subterranean clover pasture had no impact on the tenderness, juiciness, and flavor of broiler meat, members of a 30-person consumer panel classified the meat from grazing broilers with greater scores for overall appreciation. Together, the results suggest that pasture intake promotes bird performance while contributing to the production of broiler meat with preferred sensory attributes.

Influence of pasture intake on the fatty acid composition, and cholesterol, tocopherols, and tocotrienols content in meat from free-range broilers.

Over the last centuries, Western diets acquired a dramatic imbalance in the ratio of polyunsaturated fatty acids (PUFA) to saturated fatty acids (SFA) with a concomitant reduction in the dietary proportion of n-3 PUFA. Pastures are a good source of n-3 fatty acids, although the effect of forage intake in the fatty acid profile of meat from free-range chicken remains to be evaluated. In addition, it is unknown if consumer interest in specialty poultry products derived from free-range or organic production systems is accompanied by a greater nutritional quality of these products. In this study, broilers of the RedBro Cou Nu x RedBro M genotype were fed on a cereal-based diet in portable floorless pens located either on subterranean clover (Trifolium subterraneum) or white clover (Trifolium repens) pastures. Control birds were maintained at the same site in identical pens but had no access to pasture. The capacity of ingested forage to modulate broiler meat fatty acid profiles and the meat content of total cholesterol, tocopherols, and tocotrienols was investigated in broiler chicks slaughtered at d 56. The results suggested that pasture intake (<5% DM) had a low impact on the fatty acid and vitamin E homologue profiles of meat from free-range broilers. However, breast meat from birds with free access to pasture presented lower levels of the n-6 and n-3 fatty acid precursors linoleic acid (18:2n-6) and alpha-linolenic acid (18:3n-3), respectively. In spring the levels of eicosapentaenoic acid (20:5n-3) in breast meat were significantly greater in birds consuming pastures, which suggests greater conversion of alpha-linolenic acid into eicosapentaenoic acid in these birds. Finally, when compared with meat from slower-growing genotypes obtained under the conventional European free-range production systems with slaughtering at d 81, meat from birds of the Ross genotype raised intensively and slaughtered at d 35 seemed to have greater nutritional quality.