Chinese Journal of Organic Chemistry ›› 2019, Vol. 39 ›› Issue (12): 3490-3497.DOI: 10.6023/cjoc201906008 Previous Articles     Next Articles


郭兵博, 沈生强, 金淑惠, 路慧哲, 张建军   

  1. 中国农业大学理学院 北京 100193
  • 收稿日期:2019-06-10 修回日期:2019-07-31 发布日期:2019-08-07
  • 通讯作者: 金淑惠, 张建军;
  • 基金资助:

Synthesis and Enzyme Substrate Activity of p-Nitrophenyl N,N'-Diacetyl-4-thio-β-chitobioside

Guo Bingbo, Shen Shengqiang, Jin Shuhui, Lu Huizhe, Zhang Jianjun   

  1. School of Science, China Agricultural University, Beijing 100193
  • Received:2019-06-10 Revised:2019-07-31 Published:2019-08-07
  • Supported by:
    Project supported by the National Natural Science Foundation of China (No. 21772230) and the Chinese Universities Scientific Fund (No. 2019TC135).

Based on the resistance of glycosidase to the hydrolysis of glycosidase, the probe molecule p-nitrophenyl N,N'-diacetyl-4-thio-β-chitobioside (pNP-TCB) was designed and synthesized. The effects of pNP-TCB on the degradation of glycoside hydrolase family 18 (GH18) chitinase and 20 (GH20) β-N-acetylhexosaminidase were studied. The results showed that when pNP-TCB was combined with β-N-acetylhexosaminidase, the disaccharide substrate was hardly degraded. But when it reacted with chitinase, the pNP-TCB was degraded by enzyme to release p-nitrophenol group, and the corresponding ultraviolet absorption value can be detected by enzyme-labeled instrument at 405 nm. It indicates that the substrate molecule exhibits good specificity, which can be used to study the properties of GH18 chitinase at the cellular level without interference of β-N-acetylhexosaminidase. The pNP-TCB can also provide a good detection method for screening and isolation between the GH18 chitinase and the GH20 β-N-acetyl hexose.

Key words: p-nitrophenol, disaccharide substrate, chitobioside, chitinase, UV absorption