在大豆品种中测定 kunitz 胰蛋白酶抑制剂时，芯片实验室方法的优势。

Advantages of the lab-on-a-chip method in the determination of the kunitz trypsin inhibitor in soybean varieties.

机构信息

Institute for Food Technology, University of Novi Sad, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia.

出版信息

J Agric Food Chem. 2010 Jul 14;58(13):7980-5. doi: 10.1021/jf100830m.

PMID:20527986

Abstract

Qualitative and quantitative determination of the Kunitz trypsin inhibitor (KTI) in soybean experimental lines is very important in processes of selecting and breeding of new varieties. The total enzyme activity assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and Lab-on-a-Chip (LoaC) method were used for the determination of the presence and quantity of KTI in 15 soybean experimental lines and varieties. From the total trypsin inhibitor enzyme assay, inhibitor activities were registered in all samples, even in a Kunitz variety that was a negative control. The SDS-PAGE method did not detect the presence of the KTI protein band in seven soybean experimental lines and Kunitz variety, while the LoaC method showed the absence of KTI only in the Kunitz variety sample. Results confirmed the superiority of the LoaC method over other two methods in selectivity and sensitivity when KTI determination is concerned. Relationships between the KTI content obtained by the LoaC method and total trypsin inhibitor enzyme activity were established and statistically confirmed.

摘要

定性和定量测定大豆实验品系中的 Kunitz 胰蛋白酶抑制剂（KTI）在品种选择和培育过程中非常重要。总酶活性测定、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和片上实验室（LoaC）方法用于测定 15 种大豆实验品系和品种中 KTI 的存在和数量。从总胰蛋白酶抑制剂酶测定中，所有样品中都检测到了抑制剂活性，甚至在作为阴性对照的 Kunitz 品种中也是如此。SDS-PAGE 方法未在 7 种大豆实验品系和 Kunitz 品种中检测到 KTI 蛋白带的存在，而 LoaC 方法仅在 Kunitz 品种样品中显示不存在 KTI。结果证实，在测定 KTI 时，LoaC 方法在选择性和灵敏度方面优于其他两种方法。建立了 LoaC 方法获得的 KTI 含量与总胰蛋白酶抑制剂酶活性之间的关系，并通过统计学方法得到了证实。