High Levels Of Chitinase Research Articles

Chitotriosidase and lysosomal enzymes as potential biomarkers of disease progression in amyotrophic lateral sclerosis: A survey clinic-based study

The aim of this study was to determine if blood chitotriosidase (Chit) activity and lysosomal enzyme levels might represent markers of disease activity and progression in amyotrophic lateral sclerosis (ALS).It is a survey clinic-based study performed in a tertiary ALS centre. Blood samples were obtained from 76 patients with ALS in different stages of the disease and from 106 healthy individuals serving as controls. Chit activity and the levels of acid alpha-glucosidase, acid alpha-galattosidase A, beta-glucocerebrosidase, and alpha-l-iduronidase were detected using the dried blood spots (DBS) technique. The CHIT1 genotype for exon 10 duplication and for the p.G102S variant was also determined. Chit activity was significantly higher in ALS patients than in healthy individuals. This difference was independent of the genotypes at CHIT1 functional variants. Chit were significantly higher in 34 rapidly progressing patients as compared to 42 with slowly progressive disease. Acid alpha-glucosidase was higher than normal and significantly correlated with the severity of the disease. Glucocerebrosidase and alpha-l-iduronidase activity were significantly lower in patients than in the controls. Alpha-galactosidase A was higher than normal only in rapidly progressing patients.We have employed a very simple and affordable laboratory test to measure blood Chit and lysosomal enzymes activity which could be easily included in the screening of ALS patients recruited in clinical trials. Remarkably, high levels of chitinase and alpha-galactosidase A could help to distinguish patients with fast progression from those with slow progression of the disease and possibly to follow the effects of treatments on neuroinflammation and autophagy.

Digestive enzymes in larvae of the leaf cutting ant, Acromyrmex subterraneus (Hymenoptera: Formicidae: Attini)

The digestive physiology and biochemistry of larvae of the leaf-cutting ant Acromyrmex subterraneus were investigated here. The activity of digestive enzymes was evaluated in the labial glands, midgut epithelium (soluble and particulate fractions), and in the lumen contents, separated into endo and ectoperitrophic regions. Enzymes with high levels of activity were partially characterised using chromatography and electrophoresis techniques. Microscope observations were carried out and the anatomy of the larval digestive tract was described here for the first time. Larvae fed with pH indicator solutions showed the anterior portion of the midgut to be acidic and the posterior portion neutral to alkaline, indicating that the pH of the different regions of the midgut could optimise certain enzyme activities, whilst inhibiting others. The flow rate of the intestinal contents was also evaluated in larvae fed with a dye solution. The slow flow rate is probably due to closure of the rear end of the larval midgut. No compartmentalisation of digestive enzymes acting on oligosaccharides and disaccharides in the ectoperitrophic space and on polysaccharides in the endoperitrophic space was observed here, which could also be related to the closure of the midgut. The digestive physiology of these larvae is therefore similar to ancestral Holometabola, a paradox when considering the highly evolved nature of these insects. The larval midgut demonstrated a large diversity of enzyme activities with high levels of α-amylase, α-mannosidase, chitinase, α-glucosidase, β-glucosidase and proteinase. High levels of chitinase and amylase activities were detected in the labial glands of larvae. The enzyme profile reflected the necessity of the larvae to degrade the fungal substrate, their sole diet, and a probable source of some of the digestive enzymes detected here. When compared to adults, the larvae had a greater diversity and higher levels of enzyme activity, highlighting their importance as the “digestive caste” of the colony.

Chitinase and β-1,3-glucanase enzymatic activities in response to infection by Alternaria alternata evaluated in two stages of development in different tomato fruit varieties

Chitinase (ChA) and β-1,3-glucanase (GA) activity had been related with plant defense mechanism against pathogen attack in vegetative tissues. Scarce information is available about the behavior of these enzymes in response to different stages of development and fungi infection in fruits. The changes in ChA and GA activities in response to Alternaria alternata infection were evaluated in mature green (MG) and red ripe (RR) developmental stages of Sunpride, Geronimo and Charleston varieties of tomato fruit. Tomato fruits were inoculated with a conidial suspension of A. alternata and stored for 10 days at 25 °C and 90–92 H.R.%. The degree of fruit infection was measured by a hedonic scale every 2 days. ChA activity was determined fluorometrically by quantifying the release of 4-methylumbelliferyll (4-MU) from 4-methylumbelliferyll β-d-N,N′N″-triacetylchitotrioside, and GA activity was measured quantifying the release of glucose from β-1,3-glucan (laminarin) by HPLC. Tomato fruit in RR stage was more susceptible to fungi infection than MG stages. Geronimo was the most resistant variety, whereas Sunpride was the most susceptible for both stages of development (MG and RR). Higher levels of ChA and GA activities were observed for mature green stage in Charleston variety at the end of the storage period. An induction in ChA and GA in response to infection by A. alternata was observed in all varieties. Particularly high levels of ChA were found for inoculated Geronimo in RR stage and inoculated Charleston in MG stage which correlated with low levels of fungi infection. Higher levels of GA induction in response to fungi infection were recorded for Sunpride variety in RR stage, whereas no substantial induction was observed for Geronimo and Charleston varieties at the same stage of development. This GA induction correlated negatively with the resistance showed by the different varieties to fungi infection. We concluded that chitinase and β-1,3-glucanase induction are part of the tomato fruit defense mechanism against A. alternata infection with a different behavior depending upon stage of development and variety.

Optimization of cultivation conditions for the production of chitinase from Paenibacillus sp CHE-N1

A bacterial strain capable of secreting high levels of chitinase was isolated from chitin-enriched soil and identified as Paenibacillus sp. CHE-N1. This strain, which shows high potential as an antagonist against fungi, was grown in a chitin-containing medium to evaluate its ability to produce chitinase. The statistical design method was used to determine the optimal chitinase-producing conditions. Factors, including carbon sources and nitrogen sources in the medium compositions, initial pH, and the temperature of fermentation, were investigated. The obtained optimal conditions were: CSCP 55.4g/L, peptone 1.8g/L, and (NH4)2SO4 1.6g/L, with an optimal initial pH and temperature of 8.5 and 34.3℃, respectively. Under these conditions, the optimal volumetric enzyme activity could reach 8.1U/mL, about a 4-fold increase as compared to results under basic conditions.

An endophytic chitinase-producing isolate of Actinoplanes missouriensis, with potential for biological control of root rot of lupin caused by Plectosporium tabacinum

Twenty-one streptomycete and 15 non-streptomycete actinomycetes were isolated from surface-disinfested lupin roots and evaluated for their potential to produce chitinase and to inhibit the growth of Plectosporium tabacinum, the causal agent of lupin root rot in Egypt. The most inhibitory isolate was identified as Actinoplanes missouriensis which produced relatively high levels of chitinase and degraded the hyphae of P.�tabacinum in vitro, causing extensive plasmolysis and cell-wall lysis. A crude culture filtrate of A. missouriensis exhibited antifungal activity and significantly (P < 0.05) reduced spore germination and germ-tube growth of the pathogen. The antagonist was recovered from inside the root at all samplings up to 8 weeks after inoculation, indicating that the roots of healthy lupin may be a habitat for the endophyte. A. missouriensis significantly (P < 0.05) reduced the severity of root rot under glasshouse conditions. An endophytic isolate of Actinoplanes italicus incapable of producing chitinase and a mutant strain of A. missouriensis that did not produce detectable levels of chitinase, did not lyse hyphae of P. tabacinum or reduce root rot in the glasshouse experiments, although colonisation of the lupin root by both these isolates was similar to that of the chitinase-producing wild-type isolate of A. missouriensis. This study is the first record of control of a soil-borne plant pathogen by a chitinolytic actinomycete, endophytic in plant roots.

High-level expression of a tobacco chitinase gene in Nicotiana sylvestris. Susceptibility of transgenic plants to Cercospora nicotianae infection

Endochitinases (E.C.3.2.14, chitinase) are believed to be important in the biochemical defense of plants against chitin-containing fungal pathogens. We introduced a gene for class I (basic) tobacco chitinase regulated by Cauliflower Mosaic Virus 35S-RNA expression signals into Nicotiana sylvestris. The gene was expressed to give mature, enzymatically active chitinase targeted to the intracellular compartment of leaves. Most transformants accumulated extremely high levels of chitinase-up to 120-fold that of non-transformed plants in comparable tissues. Unexpectedly, some transformants exhibited chitinase levels lower than in non-transformed plants suggesting that the transgene inhibited expression of the homologous host gene. Progeny tests indicate this effect is not permanent. High levels of chitinase in transformants did not substantially increase resistance to the chitin-containing fungus Cercospora nicotiana, which causes Frog Eye disease. Therefore class I chitinase does not appear to be the limiting factor in the defense reaction to this pathogen.

Activities of chitinase and protease and concentration of fluoride in the digestive tract of Antarctic fishes feeding on krill ( Euphausia superba Dana)

1. 1. Studies on the digestion of krill by Notothenia rossii marmorata, Notothenia neglecta, Champsocephalus gunnari and Chaenocephalus aceratus showed that these Antarctic fish species are well equipped to feed on krill, as indicated by their high levels of chitinase and protease activity. 2. 2. Very high chitinolytic activities were determined in the stomach of the fish species. However, activities that were measured in intestine samples can be substantial, as well. Very strong protease activities were determined in samples of the stomach tissue and the intestinal contents. 3. 3. When krill were present in the guts, the concentrations of fluoride in the stomach and intestinal contents of N. rossii marmorata and Ch. gunnari were extremely high, while the tissues were practically devoid of fluoride.