Data/hora: 09/06/2024 02:31:26

Biblioteca(s): | Embrapa Caprinos e Ovinos.
Data corrente: | 01/08/1992
Data da última atualização: | 26/04/2023
Autoria: | IRITANI, A.; NISHIKAWA, Y.
Afiliação: | Lab. of Animal Reprod., College of Agric., Kyoto Univ.; Lab. of Animal Reprod., College of Agric., Kyoto Univ.
Título: | Studies on the egg yolk-coagulating enzyme in goat semen. V. Purification of the egg yolk coagulating enzyme.
Ano de publicação: | 1963
Fonte/Imprenta: | Japanese Journal of Animal Reproduction, v. 8, n. 4, p. 118-121, 1963.
DOI: | https://doi.org/10.1262/jrd1955.8.118
Idioma: | Japonês
Conteúdo: | Abstract: Existence of the egg yolk-coagulating enzyme in goat semen was found recently, yet its purificationhas not been attempted. While it is generally very important and significant to extract an enzyme inthe purified state in order to clarify the characteristics especially for those discovered such as the onediscussed here. In this experiment, the authors tried to purify the enzyme by the following methods; salting out byammonium sulfate, selecting adsorbtion with tricalcium phosphate-gel, andthe acetone fractionation. Principal results obtained were as follows. 1. Selecting precipitation method with ammonium sulfate was tried on the goat seminal plasma toobtain the enzyme rich fraction, but most ofthe seminal plasma protein was abruptly salted out ataround 30%ammonium sulfate. And it was difficult to extract the enzyme rich fraction by this method. 2. Then the selecting adsorbtion method was performed. Seminal plasma protein was salted out adding equal volume of saturated ammonium sulfate, and dialized the precipitated protein for 12 hours at 4°C, then it was adsorbed to the tricalcium phosphate-gel, and the selecting elution was made addingvarious concentrations of the phosphate buffer to the protein adsorbed calcium-gel. As the results, considerably purified enzyme was obtained from the fraction eluted into 0.25 M phoshate buffer. Besides, the purity of the enzyme in this fraction was about 10 times higher than that of the starting material, seminal plasma. 3. Purification of the coagulating enzyme was performed by the acetone fractionation starting from the goat seminal plasma. Acetone was added to the goat seminal plasma to be 33%, 45%, 51%, 62.%, 67%, and 77% of the concentration in the mixture at 0°C, then each of the precipitated fraction wasobtained after ultracentrifugation. Thus, it was confirmed that the fraction precipitated at the concentration of 33%acetone shows the highest apecific scivity, and the purity of the enzyme was about 4 times higher than that of the starting materia1, goat seminal plasma. Furthermore, paperelectrophoretic studies were performed on each of the fraction obtained above. Andit was clarified that the enzyme-rich fraction shows low mobility, and the remaining 4 fractions free from most of the enzyme have high mobility.
Thesagro: | Bioquímica; Caprino; Coagulação; Enzima; Reprodução; Sêmen.
Thesaurus Nal: | Animal reproduction; Egg yolk; Enzymes; Goats.
Categoria do assunto: | L Ciência Animal e Produtos de Origem Animal
Registro original: | Embrapa Caprinos e Ovinos (CNPC)
Exemplares: | CNPC - (9875-1) - ADD - AP - DD -  -  -  - DIS