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1.
J Nutr ; 137(7): 1795-801, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17585033

RESUMO

Young pigs represent an excellent model of youth to assess potentials of dietary factors for improving bone structure and function. We conducted 2 experiments to determine whether adding microbial phytase (2,000 U/kg, OptiPhos, JBS United) and Sr (50 mg/kg, SrCO3 Alfa Aesar) into a P-adequate diet further improved bone strength of young pigs. In Expt. 1, 24 gilts (8.6 +/- 0.1 kg body wt) were divided into 2 groups (n = 12), and fed a corn-soybean-meal basal diet (BD, 0.33% available P) or BD + phytase for 6 wk. In Expt. 2, 32 pigs (11.4 +/- 0.2 kg) were divided into 4 groups (n = 8), and fed BD, BD + phytase, BD + Sr, or BD + phytase and Sr for 5 wk. Both supplemental phytase and Sr enhanced (P < 0.05) breaking strengths (11-20%), mineral content (6-15%), and mineral density (6-11%) of metatarsals and femurs. Supplemental phytase also resulted in larger total bone areas (P < 0.05) and a larger cross-sectional area of femur (P = 0.06). Concentrations of Sr were elevated 4-fold (P < 0.001) in both bones by Sr, and moderately increased (P = 0.05-0.07) in metatarsal by phytase. In conclusion, supplemental phytase at 2000 U/kg of P-adequate diets enhanced bone mechanical function of weanling pigs by modulating both geometrical and chemical properties of bone. The similar benefit of supplemental Sr was mainly due to an effect on bone chemical properties.


Assuntos
6-Fitase/metabolismo , Densidade Óssea/efeitos dos fármacos , Escherichia coli/enzimologia , Fósforo na Dieta/farmacologia , Estrôncio/farmacologia , Suínos/crescimento & desenvolvimento , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Suplementos Nutricionais
2.
J Nutr ; 136(12): 3033-8, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17116716

RESUMO

Iron deficiency represents one of the most common global nutritional disorders in humans. Our objective was to determine whether and how supplemental inulin improved utilization of iron intrinsically present in a corn and soybean meal diet by young pigs for hemoglobin repletion. In Expt. 1, 3 groups (n = 8/group) of pigs were fed a corn and soybean meal-based diet (BD, without inorganic iron addition) or BD + 2 or 4% inulin (Synergy 1: a mixture of oligofructose and long-chain inulin HP, Orafti) for 5 wk. Final blood hemoglobin concentrations and the overall hemoglobin repletion efficiency of pigs were positively (r = 0.55 and 0.69, P < 0.01) correlated with dietary inulin concentrations. Compared with pigs fed the BD, those fed 4% inulin demonstrated a 28% improvement (P < 0.01) in hemoglobin repletion efficiency and 15% (P < 0.01) improvement in the final blood hemoglobin concentration. In Expt. 2, 12 weanling pigs (n = 6/group) were fed the BD or the BD + 4% inulin for 6 wk. Pigs fed 4% inulin had higher (P < 0.05) soluble Fe concentrations in the digesta of the proximal, mid, and distal colon, and lower (P < 0.05) sulfide concentrations in the digesta of the distal colon. Supplemental inulin had virtually no effect on pH or phytase activity of digesta from any of the tested segments. In conclusion, supplementing 4% inulin improved utilization of intrinsic iron in the corn and soybean meal diet by young pigs, and this benefit was associated with soluble Fe and sulfide concentrations but not pH or phytase activity in the digesta.


Assuntos
Ração Animal , Suplementos Nutricionais , Glycine max , Insulina/administração & dosagem , Ferro/metabolismo , Zea mays , Animais , Disponibilidade Biológica , Hormônio do Crescimento/sangue , Insulina/farmacologia , Modelos Animais , Suínos , Desmame
3.
Appl Environ Microbiol ; 72(6): 4397-403, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16751556

RESUMO

Environmental pollution by phosphorus from animal waste is a major problem in agriculture because simple-stomached animals, such as swine, poultry, and fish, cannot digest phosphorus (as phytate) present in plant feeds. To alleviate this problem, a phytase from Aspergillus niger PhyA is widely used as a feed additive to hydrolyze phytate-phosphorus. However, it has the lowest relative activity at the pH of the stomach (3.5), where the hydrolysis occurs. Our objective was to shift the pH optima of PhyA to match the stomach condition by substituting amino acids in the substrate-binding site with different charges and polarities. Based on the crystal structure of PhyA, we prepared 21 single or multiple mutants at Q50, K91, K94, E228, D262, K300, and K301 and expressed them in Pichia pastoris yeast. The wild-type (WT) PhyA showed the unique bihump, two-pH-optima profile, whereas 17 mutants lost one pH optimum or shifted the pH optimum from pH 5.5 to the more acidic side. The mutant E228K exhibited the best overall changes, with a shift of pH optimum to 3.8 and 266% greater (P < 0.05) hydrolysis of soy phytate at pH 3.5 than the WT enzyme. The improved efficacy of the enzyme was confirmed in an animal feed trial and was characterized by biochemical analysis of the purified mutant enzymes. In conclusion, it is feasible to improve the function of PhyA phytase under stomach pH conditions by rational protein engineering.


Assuntos
6-Fitase/genética , 6-Fitase/metabolismo , Ração Animal , Aspergillus niger/enzimologia , Aditivos Alimentares , Concentração de Íons de Hidrogênio , 6-Fitase/química , Substituição de Aminoácidos , Sítios de Ligação , Poluição Ambiental , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Cinética , Modelos Moleculares , Mutagênese Sítio-Dirigida , Conformação Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo
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