三角酵母天门冬氨酸氧化酶的诱导。
Induction of the d-Amino Acid Oxidase from Trigonopsis variabilis.
出版信息
Appl Environ Microbiol. 1996 Jun;62(6):2106-10. doi: 10.1128/aem.62.6.2106-2110.1996.
Induction of the d-amino acid oxidase (EC. 1.4.3.3) from the yeast Trigonopsis variabilis was investigated by using a minimal medium containing glucose as the carbon and energy source, (NH(inf4))(inf2)SO(inf4) as the nitrogen source, and various d- and dl-amino acid derivatives as inducers. The best new inducers found were N-carbamoyl-d-alanine, N-acetyl-d-tryptophan, and N-chloroacetyl-d-(alpha)-aminobutyric acid; when the induction effects of these compounds were compared with the effects of d-alanine as the nitrogen source and inducer, the resulting activities of d-amino acid oxidase per gram of dried yeast were 4.2, 2.1, and 1.5 times higher, respectively. The optimum concentration of the best inducer, N-carbamoyl-d-alanine, was 5 mM. This inducer could also be used in its racemic form. The induction was pH dependent. After cultivation of the yeast in a 50-liter bioreactor, d-amino acid oxidase activity of about 3,850 (mu)kat (231,000 U) was obtained. In addition, production of the d-amino acid oxidase was found to be significantly dependent on the metal salt composition of the medium. Addition of zinc ions was required to obtain high d-amino acid oxidase levels in the cells. The optimum concentration of ZnSO(inf4) was about 140 (mu)M.
我们考察了利用以葡萄糖为碳源和能源、(NH₄)₂SO₄为氮源、各种 D-和 DL-氨基酸衍生物为诱导物的最小培养基，从酵母三角酵母（Trigonopsis variabilis）中诱导 D-氨基酸氧化酶（EC.1.4.3.3）的情况。发现的最佳新型诱导物为 N-氨甲酰-D-丙氨酸、N-乙酰-D-色氨酸和 N-氯乙酰-D-（α）-氨基丁酸；将这些化合物的诱导效果与 D-丙氨酸作为氮源和诱导物的效果进行比较，每克干酵母的 D-氨基酸氧化酶的活性分别高出 4.2、2.1 和 1.5 倍。最佳诱导物 N-氨甲酰-D-丙氨酸的最佳浓度为 5mM。该诱导物也可以使用其外消旋形式。诱导作用依赖于 pH 值。在 50 升生物反应器中培养酵母后，获得了约 3850（μ）kat（231000U）的 D-氨基酸氧化酶活性。此外，发现 D-氨基酸氧化酶的产生显著依赖于培养基中的金属盐成分。需要添加锌离子才能使细胞中获得高 D-氨基酸氧化酶水平。ZnSO₄的最佳浓度约为 140μM。
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