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Characterization of serine proteinase inhibitors in dry seeds of cultivated pasture grass species

Published online by Cambridge University Press:  19 September 2008

Mohammed Tasneem ,
Clive A. Cornford  and
Michael T. McManus
Mohammed Tasneem
Affiliation:
Plant Molecular Genetics Laboratory, Grasslands Research Centre, AgResearch, Private Bag 11008, Palmerston North, New Zealand. Department of Plant Biology and Biotechnology, Massey University, Private Bag, Palmerston North, New Zealand
Clive A. Cornford
Affiliation:
Department of Plant Biology and Biotechnology, Massey University, Private Bag, Palmerston North, New Zealand
Michael T. McManus*
Affiliation:
Plant Molecular Genetics Laboratory, Grasslands Research Centre, AgResearch, Private Bag 11008, Palmerston North, New Zealand.
*
*Correspondence
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Abstract

A survey of proteinaceous inhibitors of the serine proteinases, bovine trypsin and chymotrypsin, that are extractable from dry seeds of several cultivars of pasture grasses has been undertaken. Using crude extracts, most cultivars screened contained inhibitors of chymotrypsin, whereas trypsin inhibition was not detectable. Seeds from four cultivars, Lolium perenne L. cv. Grasslands Ruanui, Lolium × boucheanum cv. Grasslands Greenstone, Festuca arundinacea Schreb. cultivars Grasslands Roa and Grasslands Garland, that contained more potent chymotrypsin inhibition were purified further. After gel filtration chromatography, both trypsin and chymotrypsin inhibition could be observed in all four cultivars, and each separated into two discrete native molecular weights; one of ca. 20–22 kDa and one of ca. 8–10 kDa. However, activity staining, after polyacrylamide gel electrophoresis, revealed an array of iso-inhibitors with molecular weights that ranged from ca. 3 kDa to 20 kDa. One of these, a dual trypsin/chymotrypsin inhibitor of ca. 12 kDa that is present in all four cultivars examined, was purified to homogeneity from F. arundinacea cv. Grasslands Garland using anhydro-trypsin affinity chromatography and reverse-phase HPLC. The protein was found to comprise two closely related peptides and N-terminal amino acid sequencing revealed highest identity with a trypsin inhibitor identified in rye (Secale cereale) seeds.

Keywords

Festuca arundinacea SchrebLolium perenne Lgrass seedsprotein purificationserine proteinase inhibitors
Type
Research Papers
Information
Seed Science Research , Volume 4 , Issue 3 , September 1994 , pp. 335 - 345
Copyright
Copyright © Cambridge University Press 1994

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