ANTIBACTERIAL ACTIVITY OF A TRYPSIN-CHYMOTRYPSIN-ELASTASE INHIBITOR ISOLATED FROM LAVATERA CASHMERIANA CAMB. SEEDS

ABSTRACT Protease inhibitors were extracted from the seeds of Lavatera cashmeriana Camb by ammonium sulphate precipitation and purified by chromatography on DEAE-cellulose and Sephadex G-100 column. The bound protein eluted into four major peaks which we named as LC-pi I, II, III and IV, all showing strong anti protease activity against trypsin, chymotrypsin and elastase. LC-pi I was screened for antibacterial activity against Klebsiella pnuemoniae, Escherichia coli and Pseudomonas aeruginosa. The protease inhibitor showed strong antibacterial activity against Klebsiella pnuemoniae and Pseudomonas aeruginosa but was less active against Escherichia coli

Endonuclease heteroduplex mismatch cleavage for detecting mutation genetic variation of trypsin inhibitors in soybean

The objective of this work was to evaluate the genetic variation of trypsin inhibitor in cultivated (Glycine max L.) and wild (Glycine sofa Siebold & Zucc.) soybean varieties. Genetic variations of the Kunitz trypsin inhibitor, represented by a 21-kD protein (KTI), and of the Bowman-Birk trypsin chymotrypsin inhibitor (BBI) were evaluated in cultivated (G. max) and wild (G. sofa) soybean varieties. Endonuclease heteroduplex mismatch cleavage assays were performed to detect mutations in the KTI gene, with a single-stranded specific nuclease obtained from celery extracts (CEL I). The investigated soybean varieties showed low level of genetic variation in KTI and BBI. PCR-RFLP analysis divided the BBI-A type into subtypes A1 and A2, and showed that Tib type of KTI is the dominant type. Digestion with restriction enzymes was not able to detect differences between ti-null and other types of Ti alleles, while the endonuclease heteroduplex mismatch cleavage assay with CEL I could detect ti-null type. The digestion method with CEL I provides a simple and useful genetic tool for SNP analysis. The presented method can be used as a tool for fast and useful screening of desired genotypes in future breeding programs of soybean