Abstract
Metallo-proteinase from 8-d-old seedlings of kale was isolated. The enzyme was extracted with 1% NaCl, concentrated by ammonium sulfate and finally purified by high-performance liquid chromatography. The isolated enzyme had a molecular weight of 22.4 kDa and showed a maximum activity at pH 9.0 using casein as a substrate. Proteolytic activity of proteinase was inhibited by chelators. The inhibition by ethylenediaminetetraacetate (EDTA) was abolished by some divalent metals ions, especially by Zn2+. The enzyme showed activity against the synthetic peptides Suc-Ala-Ala-Pro-Leu-pNA and Suc-Ala-Ala-Pro-Phe-pNA, and hydrolized the following peptide bonds in the oxidized insulin B-chain: Leu6-Cya7, Leu15-Tyr16, Leu17—Val18 and Phe25-Tyr26.
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Abbreviations
- EDTA:
-
ethylenediaminotetraacetic acid
- HPLC:
-
high-performance liquid chromatography
- NEM:
-
N-ethylmaleimide
- PCMB:
-
p-mecuribenzoic acid
- PMSF:
-
phenylmethylsulfonyl fluoride
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This work was supported by the University Science Programme, Ministry of National Education, and Polish Academy of Science, Warsaw, Poland.
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Wilimowska-Pelc, A., Dryjański, M., Żal, T. et al. Metallo-proteinase from the seedlings of kale (Brassica oleracea L. var. sabellica):. Planta 185, 344–349 (1991). https://doi.org/10.1007/BF00201054
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DOI: https://doi.org/10.1007/BF00201054