ABSTRACT The objective of this paper was to investigate the availability of betaine present in fine wheat bran (FWB) in diets for growing pigs. We used thirty crossbred intact males, housed individually in metabolic cages and distributed in a randomized block design, with five treatments and six replicates. Blocks consisted of two periods of 15 days. Average initial body weights were 32.00±1.30 and 44.90±1.32 kg, in the first and second periods, respectively. Treatments were modified only at the level of methionine plus cysteine (Met+Cys) required and the Met+Cys sources, consisting of a negative control (NC), formulated with 80% of Met+Cys requirements, and four positive controls (PC): PCMet - NC supplemented with 1.2 g kg−1 of DL-methionine; PCBet - NC supplemented with 0.9 g kg−1 of anhydrous betaine; PCFWB - diet formulated with FWB; and PCFWB+Met - PCFWB supplemented with 1.0 g kg−1 of DL-methionine. No differences were observed on performance-related variables or on the digestibility coefficient of dry matter. Conversely, the digestibility coefficient of crude energy was lower when pigs were fed FWB and could be associated to the higher fiber content of FWB feeds. Digestible energy of feeds was higher for FWB diets, while the metabolizable energy of FWB diets did not differ from PCMet and PCBet. Nitrogen intake and apparent absorption were lower when pigs were fed FWB, but nitrogen retention, protein utilization, and biological value of the feed protein were the same among PC treatments, indicating that nitrogen from diets was used by pigs with the same efficiency, and is supported by no differences in performance-related variables. The FWB composition showed 12 g of betaine kg−1, which is available to maintain nitrogen retention and performance.
Apesar de possuir uma família de proteínas de defesa, o feijão-comum (Phaseolus vulgaris L.) pode ser atacado por insetos bruquídeos causando sérios danos aos grãos armazenados. O P. vulgaris possui duas formas ativas de inibidores de a-amilases, denominadas a-AI1 e a-AI2, que apresentam diferentes especificidades em relação às a-amilases. A a-amilase de Zabrotes subfasciatus é inibida por a-AI2 mas não por a-AI1. Em contraste, a a-amilase pancreática de porco é inibida por a-AI1 mas não é por a-AI2. O objetivo deste trabalho foi entender as bases moleculares da especificidade desses inibidores em relação às a-amilases. Para tanto, foram construídos mutantes do a-AI2, os quais foram expressados em plantas de fumo. Todos os inibidores mutantes deixaram de inibir a a-amilase de inseto sem, contudo, passar a exibir atividade contra a a-amilase de mamífero. Os modelos estruturais explicam por que a substituição de His33 do a-AI2 pela seqüência correspondente do a-AI1 (Ser-Tyr-Asn) aboliu a inibição da a-amilase de Z. subfasciatus. Dos estudos de modelagem molecular pode-se concluir que o tamanho e a complexidade da interface a-amilase-inibidor explicam por que a mutação da alça N-terminal e a quebra da atividade inibitória para a-amilase de Z. subfasciatus não levam ao ganho de atividade inibitória do mutante em relação à a-amilase de porco.
Despite the presence of a family of defense proteins, Phaseolus vulgaris can be attacked by bruchid insects resulting in serious damage to stored grains. The two distinct active forms of a-amylase inhibitors, a-AI1 and a-AI2, in P. vulgaris show different specificity toward a-amylases. Zabrotes subfasciatus a-amylase is inhibited by a-AI2 but not by a-AI1. In contrast, porcine a-amylase is inhibited by a-AI1 but not by a-AI2. The objective of this work was to understand the molecular basis of the specificity of two inhibitors in P. vulgaris (a-AI1 and a-AI2) in relation to a-amylases. Mutants of a-AI2 were made and expressed in tobacco plants. The results showed that all the a-AI2 mutant inhibitors lost their activity against the insect a-amylases but none exhibited activity toward the mammalian a-amylase. The replacement of His33 of a-AI2 with the a-AI1-like sequence Ser-Tyr-Asn abolished inhibition of Z. subfasciatus a-amylase. From structural modeling, the conclusion is that the size and complexity of the amylase-inhibitor interface explain why mutation of the N-terminal loop and resultant abolition of Z. subfasciatus a-amylase inhibition are not accompanied by gain of inhibitory activity against porcine a-amylase.