Method for assaying activity of prophenoloxidase activating enzyme and application thereof
申请人:Wako Pure Chemical Industies, Ltd.
公开号:EP0657546A1
公开(公告)日:1995-06-14
A method for assaying an activity of a prophenoloxidase activating enzyme (PPAE), comprising assaying at least X-Arg or Y produced upon contact of the PPAE with a peptide chain of the formula
X - Arg - Y
wherein X is an optionally labeled amino acid residue or peptide residue, having an optionally protected α-amino group or N-terminal, provided that the amino acid residue adjoining Arg is not Gly or Ala, and Y is an organic residue capable of binding to a carboxyl group of Arg by acid amide bonding or ester bonding, or an optionally labeled amino acid residue or peptide residue, having an optionally protected α-carboxyl group or C-terminal, the peptide chain capable of being hydrolyzed into X-Arg and Y by a PPAE derived from an insect. According to the present invention, PPAE activity can be quantitatively assayed with precision and a highly precise method for determining β-1,3-glucan and/or peptidoglycan, wherein said PPAE activity is used as an index, can be provided. In addition, the present invention enables detection of fungi and bacteria without multiplication by culture, which makes early diagnosis of microbial infections possible. The present invention is applicable to a wide range of use such as tests for microbial contamination of water and food, safety tests of therapeutic agents such as antibiotics and injectable preparations, etc.
一种检测丙醇氧化酶活化酶(PPAE)活性的方法,包括检测 PPAE 与式中肽链接触时产生的至少 X-Arg 或 Y。
X - Arg - Y
其中 X 是可选择标记的氨基酸残基或肽残基,具有可选择保护的 α-氨基或 N-末端,条件是与 Arg 相邻的氨基酸残基不是 Gly 或 Ala,Y 是可通过酸酰胺键或酯键与 Arg 的羧基结合的有机残基、或可选择标记的氨基酸残基或肽残基,具有可选择保护的 α-羧基或 C-末端,该肽链能够被来源于昆虫的 PPAE 水解为 X-Arg 和 Y。根据本发明,可以对 PPAE 活性进行精确的定量检测,并提供一种高精度的方法来测定 β-1,3-葡聚糖和/或肽聚糖,其中所述 PPAE 活性被用作一种指标。此外,本发明无需通过培养繁殖即可检测真菌和细菌,这使得早期诊断微生物感染成为可能。本发明适用范围广泛,如检测水和食品的微生物污染、抗生素和注射制剂等治疗药物的安全性检测等。