Inhibition of chitinolytic enzymes from Streptomyces griseus (bacteria), Artemia salina (crustacea), and a cell line from Chironomus tentans (insecta) by allosamidin and isoallosamidin

Abstract

The influence of allosamidin and its diastereoisomer isoallosamidin on chitinase activity from an insect cell line of Chironomus tentans, the crustacean Artemia salina and the bacterium Streptomyces griseus was studied. The rank order of susceptibility of chitinase activity to both inhibitors was found to be: insects »crustaceans« fungi, although isoallosamidin is about 600‐fold (insect) or 400‐fold (crustacea) less active than allosamidin and does not inhibit chitinase from Streptomyces at all. The complete and exclusive inhibition of N‐acetyl‐β‐D‐glucosaminidase by an acetylgluconolactone‐derivative and chitinase activity by allosamidin indicates that both enzymes can be determined separately in crude extracts, (containing both enzymes), with the substrates 4‐methylumbelliferyl‐N‐acetylglucosaminide or 4‐methylumbelliferyl‐chitoriose, respectively.

Effect of Pentoxifylline on Chitinolytic Enzyme Activity in the Eastern Subterranean Termite (Isoptera: Rhinotermitidae)

“…The mode of inhibitory action of pentoxifylline mimics that of allosamidin on fungal, bacterial, and human chitinases (Rao et al 2005a(Rao et al , 2005b. Similar to the in vitro activity results, allosamidin and its analogue isoallosamidin were shown to significantly affect endo-chitinase activity, while having minimal to no effect on exo-chitinase activity in a Chironomus midge cell line (Spindler and Spindler-Barth 1994). Pentoxifylline treatment significantly reduced both endo-and exo-chitinase enzyme activities in vivo.…”

“…Chitin synthesis and degradation are essential metabolic pathways in arthropods, and especially important in insect inhibition of the peritrophic membrane and cuticle (Spindler and Spindler-Barth 1994). Thus, chitin metabolism has great potential as a target site for reduced-risk (towards vertebrates) insecticide development (Merzendorfer and Zimoch 2003).…”

A cDNA encoding a chitinase from the epithelial cell line ofChironomus tentans(Insecta, Diptera) and its functional expression

“…Chitinase was determined by microfluorimetric assays (Spindler and Spindler-Barth, 1994;Spindler, 1997) but was adapted for a microtitre plate fluorimeter. The test is based on the enzymatic degradation of 4-methylumbelliferyl-N,N′, N′′-triacetyl-chitotrioside (MUF-chitotriose; Calbiochem, Soden) as described by McCreath and Gooday (1992).…”

“…Moreover, substrate specificity and inhibitory properties of allosamidin and a variety of derivatives have been tested (Spindler and Spindler-Barth, 1994;Spindler et al, 1997;Spindler-Barth et al, 1998).…”

Ecdysteroid resistant subclones of the epithelial cell line from Chironomus Tentans (insecta, diptera). I. Selection and characterization of resistant clones

“…This cell line is considered as a simplified model to study early events in ecdysteroidinduced tissue differentiation (Spindler-Barth et al, 1989Spindler-Barth, 1991, 1993Lammerding-Ktippel et al, 1994) and cuticle formation. Cell differentiation and chitin metabolism are regulated by ecdysteroids (Spindler-Barth et al, 1989;Spindler and Spindler-Barth, 1994;Quack et al, 1995). Cell differentiation and chitin metabolism are regulated by ecdysteroids (Spindler-Barth et al, 1989;Spindler and Spindler-Barth, 1994;Quack et al, 1995).…”