RESUMEN
Enzymes that are used as animal feed supplements should be able to withstand temperatures of 60 to 90 degrees C, which may be reached during the feed pelleting process. The thermostability properties of three histidine acid phosphatases, Aspergillus fumigatus phytase, Aspergillus niger phytase, and A. niger optimum pH 2.5 acid phosphatase, were investigated by measuring circular dichroism, fluorescence, and enzymatic activity. The phytases of A. fumigatus and A. niger were both denatured at temperatures between 50 and 70 degrees C. After heat denaturation at temperatures up to 90 degrees C, A. fumigatus phytase refolded completely into a nativelike, fully active conformation, while in the case of A. niger phytase exposure to 55 to 90 degrees C was associated with an irreversible conformational change and with losses in enzymatic activity of 70 to 80%. In contrast to these two phytases, A. niger pH 2.5 acid phosphatase displayed considerably higher thermostability; denaturation, conformational changes, and irreversible inactivation were observed only at temperatures of >/=80 degrees C. In feed pelleting experiments performed at 75 degrees C, the recoveries of the enzymatic activities of the three acid phosphatases were similar (63 to 73%). At 85 degrees C, however, the recovery of enzymatic activity was considerably higher for A. fumigatus phytase (51%) than for A. niger phytase (31%) or pH 2.5 acid phosphatase (14%). These findings confirm that A. niger pH 2.5 acid phosphatase is irreversibly inactivated at temperatures above 80 degrees C and that the capacity of A. fumigatus phytase to refold properly after heat denaturation may favorably affect its pelleting stability.
Asunto(s)
6-Fitasa/química , Fosfatasa Ácida/química , Aspergillus fumigatus/enzimología , Aspergillus niger/enzimología , Conformación Proteica , Estructura Secundaria de Proteína , 6-Fitasa/aislamiento & purificación , 6-Fitasa/metabolismo , Fosfatasa Ácida/aislamiento & purificación , Fosfatasa Ácida/metabolismo , Estabilidad de Enzimas , Calor , Concentración de Iones de Hidrógeno , Desnaturalización Proteica , Pliegue de Proteína , TermodinámicaRESUMEN
The first 2 years' experiences with Provox prostheses at the ENT Clinic of the University of Berne are discussed. Thirty-three prostheses were inserted after primary tracheoesophageal puncture and 5 after secondary puncture. Voice success was measured with the scale of Robillard Shultz and Harrison, which encompasses not only assessment of voice quality but also frequency of use and a patient's independence in prosthesis care. After primary puncture the percentage of "functional" speakers increased progressively from 26% after 2 months to 56% after 1 year. After secondary puncture only 2 of 5 patients became functional speakers after 6 months. Because only a few of the previously laryngectomized patients who were informed about the possibility of a secondary puncture wished to receive a prosthesis, the reasons for their reservation were analyzed. Uneasiness about an additional operation and fear of complications were the most frequent motives. Further psychosocial aspects--including the unwillingness of some patients to use their fully functioning prosthesis--are discussed.