Purification, Characterization, and Antifungal Activity of Chitinases from Pineapple (Ananas comosus) Leaf

Abstract

Three chitinases, designated pineapple leaf chitinase (PL Chi)-A, -B, and -C were purified from the leaves of pineapple (Ananas comosus) using chitin affinity column chromatography followed by several column chromatographies. PL Chi-A is a class III chitinase having a molecular mass of 25 kDa and an isoelectric point of 4.4. PL Chi-B and -C are class I chitinases having molecular masses of 33 kDa and 39 kDa and isoelectric points of 7.9 and 4.6 respectively. PL Chi-C is a glycoprotein and the others are simple proteins. The optimum pHs of PL Chi-A, -B, and -C toward glycolchitin are pH 3, 4, and 9 respectively. The chitin-binding ability of PL Chi-C is higher than that of PL Chi-B, and PL Chi-A has lower chitin-binding ability than the others. At low ionic strength, PL Chi-B exhibits strong antifungal activity toward Trichoderma viride but the others do not. At high ionic strength, PL Chi-B and -C exhibit strong and weak antifungal activity respectively. PL Chi-A does not have antifungal activity.

Tissue Distribution, Synthesis Stage, and Ethylene Induction of Pineapple (Ananas comosus) Chitinases

“…6) Pineapple leaf chitinase (PL Chi)-A is an acidic class III chitinase having no antifungal activity. PL Chi-B is a weakly basic class I chitinase having strong antifungal activity.…”

“…In the pericarp, this type of enzyme was detected at all stages during fruit development. It has no antifungal activity, and the cleavage pattern of chitin oligomer by it is different from that by PL Chi-B or -C. 6) Assuming that all types of pineapple chitinases are related to a defense mechanism, one possible role of type A chitinase is to release from the cell wall of the pathogens chitin oligosaccharides, which can act as elicitors. It appears to be effective for plant defense against pathogens that type A chitinase constitutively exists in all tissues, since the resulting elicitors can induce the expression of plant defense mechanism in any tissue.…”

Molecular cloning and expression of drought-induced protein 3 (DIP3) encoding a class III chitinase in upland rice

“…One of the physiological roles of chitinases in plants is to protect plants against fungal pathogens by degrading chitin (Schlumbaum et al, 1986). Some other chitinases do not show any antifungal activities (Taira et al, 2005). Chitinases also respond to abiotic stress either in growth or developmental processes (Collinge et al, 1993;Kasprzewska, 2003;Collinge et al, 2008;Sharma et al, 2011).…”

Purification and Biochemical Characterization of a Pathogenesis Related Endochitinase Arachis hypogaea

“…Many other workers have reported multistep procedure for purification of chitinase. Ye and Ng (2005) purified a chitinase after three chromatographic steps (CM-Sephadex C-50, POROS HS and Sephadex G-75 column chromatography) from mung beans to 3.9 folds and Taira et al (2005) purified chitinase from pineapple leaf after four steps such as chitin affinity column chromatography, ButylToyopearl 650M column chromatography, Resource Q column chromatography, HPLC on Phenyl superpose column.…”

“…To date, different types of chitinases have been purified and characterized from different plants like sugar beet (El Sayed et al, 2000), soya bean (Gijzen et al, 2001), pineapple (Taira et al, 2005), pomegranate (Yang et al, 2011), and persimmon (Zhang et al, 2013).…”