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PURIFICATION AND CHARACTERIZATION OF THE CYSTEINE PROTEINASE INHIBITOR FROM MIANNONG WHEAT SEED(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
1997 02
Page:
140-147
Research Field:
Articles
Publishing date:

Info

Title:
PURIFICATION AND CHARACTERIZATION OF THE CYSTEINE PROTEINASE INHIBITOR FROM MIANNONG WHEAT SEED
Author(s):
Zeng Zhongkui Yu Yuan Zhou Hong Bao Jinku
Dept of Biology, sichuan Univ, chengdu 610064
Keywords:
cysteine proteinase inhibitorpurification of cysteine proteinase inhibitorcharacterization of cysteine proteinase inhibitor
CLC:
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DOI:
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DocumentCode:

Abstract:
Two thiol-protease inhibitors (CPI or cysteine proteinase inhibitors) from seeds of wheat (Miannong) were separeted by extraction, fraction with (NH4)2SO4,affinity chromatography on the papain-Sepharose 4B and ion-exchange chromatography on DEAE-Sepharose and gel filtration on Sephadex G-100 column. Their molecular weights estimated by SDS-PAGE or Sephadex G-100 column were 18800[CPI H for short]and 10500[CPI L]. They exhibited one hand on PAGE,SDS-PAGE and HPLC. Both CPI H and CPI L were stable at PH 2.0-11.0 and below 90℃ for 5 min. The N-terminal amino acid of CPI H was Ile. Both CPI L and CPI H could inhibit papain with ID50 7.8×10-6 mol/L and 3.4×10-6 mol/L, respectively. The inhibition of CPI L was competitive with a Ki value of 8.32×10-8 mol/L, but that of CPI H was uncompetitive. CPI H inhibited ficin with ID50 5.8×10-6 mol/L, but gave weak inhibitory effect on bromelin. CPI L also inhibited ficin with ID50 9.5×10-6 mol/L, but showed no such activity to btomelin. They could not inhibit trypsin,chymotrypsin and pepsin.

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