CgTI, a novel thermostable Kunitz trypsin-inhibitor purified from Cassia grandis seeds: Purification, characterization and termiticidal activity

International Journal of Biological Macromolecules
Romero M P Brandão-CostaAna Lucia F Porto

Abstract

Cassia grandis trypsin inhibitor (CgTI) is a novel plant serine proteinase inhibitor. This study sets out to purify a thermostable inhibitor from the seeds of Cassia grandis and to provide biochemical information about a novel peptide belonging to the Kunitz family. Moreover, toxicity assays against Artemia, Aedes aegypti larvae-L4 and Nasutitermes corniger are evaluated. The purification process was performed using acetone precipitation, Trypsin-Sepharose-CL4B and Superdex-G75. The inhibitor showed an apparent molecular mass of around 19.8 kDa on Superdex-G75 gel filtration, and a mass of around 19.0 kDa visualized by SDS-PAGE under reducing conditions, and it also showed the protein consists of two polypeptide chains. N-terminal sequencing by Edman's degradation of 16 residues revealed a sequence of amino acids SVVLDTSGEPIRNGGG. 2D-electrophoresis identified a pI value of 6.3 and a 1:1 stoichiometric ratio was noted during CgTI-trypsin complex formation. The inhibitor retained the inhibitory activity over a broad range of pH (5-10) and showed thermostable activity at temperatures 30-80 °C. Furthermore, in vivo assays showed no lethality effect against Artemia and Aedes aegypti larvae, but mortality against Nasutitermes cornig...Continue Reading

Citations

Dec 4, 2019·Bioscience, Biotechnology, and Biochemistry·Jia LiuYoshiaki Kouzuma
Feb 2, 2020·Plant Science : an International Journal of Experimental Plant Biology·Juliana CotabarrenWalter David Obregón
Jan 7, 2019·International Journal of Biological Macromolecules·Mauricio V CruzKátia F Fernandes
Jan 15, 2022·Scientific Reports·Milena do AmaralCarlos Priminho Pirovani

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