Chitin-containing Pathogens Research Articles

Biocontrol potential of cell-free supernatant of Paenibacillus chitinolyticus against Plasmodiophora brassicae in two important Brassica species

Plasmodiophora brassicae is a serious threat to Brassica crops worldwide, resulting in substantial economic losses for growers. Biocontrol with chitinolytic bacteria producing chitinase is gaining attention as a natural alternative to chemicals. This approach is favored due to the essential role chitinases play in protecting against chitin-containing pathogens. Given that chitin is a major component in the resting spores of P. brassicae and plays a crucial role during pathogenesis, it is probable that Paenibacillus chitinolyticus, producing a high level of chitinase, could suppress P. brassicae by targeting chitin in a critical stage of this pathogen’s life cycle. Our research aimed to evaluate the effect of various applications of P. chitinolyticus on clubroot suppression in two economically important Brassica species: Chinese cabbage and rapeseed. The effectiveness of the cell-free supernatant (CFS) of an endemic strain of P. chitinolyticus from the Czech Republic at five different time points was studied in the greenhouse by measuring the disease severity index. Early application of P. chitinolyticus decreased the disease index significantly within both plant species resulting in increased shoot dry weight. In conclusion, the CFS of P.chitinolyticus has significant antagonistic activity against clubroot in Chinese cabbage and rapeseed in the early developmental stages of clubroot occurrence and holds the potential as a biofertilizer as well as bioprotectant agent in clubroot management of P. brassicae.

Overexpression of chitinase PbChia1 from Plasmodiophora brassicae improves broad-spectrum disease resistance of Arabidopsis

ABSTRACT Chitinase plays an important role in plant resistance against chitin-containing pathogens through hydrolysis of chitin. Clubroot caused by Plasmodiophora brassicae is a major disease for cruciferous crops and vegetables worldwide. The cell wall of P. brassicae resting spores contains chitin. Chitinase is regarded as capable of improving plant resistance to fungal diseases. However, there has been no report about the function of chitinase in P. brassicae. Here, wheat germ agglutinin staining and commercial chitinase treatment demonstrated that chitin is a functional component in P. brassicae. In addition, Chitinase PbChia1 was identified by chitin pull-down assay combined with LC-MS/MS. PbChia1 was found to be a typical secreted chitinase, which could bind chitin with chitinase activity in vitro. PbChia1 could significantly decrease the resting spores of P. brassicae and therefore relieve the severity of clubroot symptom, with a biocontrol effect of 61.29%. Overexpression of PbChia1 in Arabidopsis thaliana improved its resistance to P. brassicae, increased host survival rate and seed yield, enhanced PAMPs-triggered reactive oxygen species burst, MAPK activation and expression of immune-related genes. PbChia1 transgenic plants also showed resistance to other pathogens, such as biotrophic bacterium Pst DC3000, necrotrophic fungi Sclerotinia sclerotiorum and Rhizoctonia solani. These findings indicate that chitinase PbChia1 is a candidate gene that can confer broad-spectrum disease resistance in breeding.

The chitinases as biomarkers in immune-mediate diseases.

The role of chitinases has been focused as potential biomarkers in a wide number of inflammatory diseases, in monitoring active disease state, and predicting prognosis and response to therapies. The main chitinases, CHIT1 and YKL-40, are derived from 18 glycosyl hydrolases macrophage activation and play important roles in defense against chitin-containing pathogens and in food processing. Moreover, chitinases may have organ- as well as cell-specific effects in the context of infectious diseases and inflammatory disorders and able to induce tissue remodelling. The CHIT1 measurement is an easy, reproducible, reliable, and cost-effective affordable assay. The clinical use of CHIT1 for the screening of lysosomal storage disorders is quite practical, when proper cut-off values are determined for each laboratory. The potential of CHIT1 and chitinases has not been fully explored yet and future studies will produce many surprising discoveries in the immunology and allergology fields of research. However, since the presence of a null CHIT1 gene in a subpopulation would be responsible of false-negative values, the assay should be completed with the other markers such ACE and, if necessary, by genetic analysis when CHIT1 is unexpected low.

Influence of chitotriosidase gene polymorphisms on oxidative stress and susceptibility to Aspergillus infection among exposed workers

ABSTRACT Chitotriosidase (CHIT1) is involved in the innate defense against chitin-containing pathogens. In the present study, we sought to investigate the role of CHIT1 gene polymorphisms on susceptibility to Aspergillus infection in addition to oxidative stress caused by infection. CHIT1 gene polymorphisms were identified in 60 Aspergillus-positive workers by REFLP. We also measured concentrations of the CHIT1 enzyme, total antioxidant capacity (TAC), and malondialdehyde (MDA). The majority of workers were wild-type (AA) (66.5%), followed by heterozygous (AB) (28.5%), and homozygous mutants (BB) (5%). The mean concentrations of specific IgE for all Aspergillus species were affected by change in CHIT1 genotypes. Our findings indicate that decreased CHIT1 activity in homozygous mutant CHIT1 allele is associated with a subsequent decrease in TAC levels, resulting in an increased risk of fungal infection and accumulation of oxidant MDA. Thus, CHIT1 enzyme activity plays a critical role in the susceptibility of WWTP workers to fungal infections.

Molecular evolution and functional characterization of chitinase gene family in Populus trichocarpa

Chitinases, the chitin-degrading enzymes, have been shown to play important role in defense against the chitin-containing fungal pathogens. In this study, we identified 48 chitinase-coding genes from the woody model plant Populus trichocarpa. Based on phylogenetic analysis, the Populus chitinases were classified into seven groups. Different gene structures and protein domain architectures were found among the seven Populus chitinase groups. Selection pressure analysis indicated that all the seven groups are under purifying selection. Phylogenetic analysis combined with chromosome location analysis showed that Populus chitinase gene family mainly expanded through tandem duplication. The Populus chitinase gene family underwent marked expression divergence and is inducibly expressed in response to treatments, such as chitosan, chitin, salicylic acid and methyl jasmonate. Protein enzymatic activity analysis showed that Populus chitinases had activity towards both chitin and chitosan. By integrating sequence characteristic, phylogenetic, selection pressure, gene expression and protein activity analysis, this study shed light on the evolution and function of chitinase family in poplar.

Transcriptional upregulation of multiple earthworm chitinase genes following bacterial challenge suggests their implications in innate immunity.

Chitinase is a multi-functional enzyme that catalyzes the hydrolysis of β-1,4-linkages between N-acetylglucosamines (GlcNAc) in chitin. Recent studies imply that earthworm chitinase is implicated in self-defense immunity against chitin-containing pathogens. However, a direct relationship of earthworm chitinase with innate immunity has not yet been established. In this study, earthworm (Eisenia andrei) chitinase expression was examined following bacterial challenge by Bacillus subtilis. RNA sequencing (RNA-seq) and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) were used to quantitatively evaluate mRNA expression changes in response to bacterial stimulation. Multiple chitinase-related mRNAs were found to be upregulated, among which EaChi3, EaChi4, and EaChi2 were upregulated by approximately eightfold, eightfold, and 2.5-fold, respectively. This strongly suggested that earthworm chitinases may act as inducible humoral effectors in earthworm innate immunity. The primary structures of all three chitinases contained an N-terminal glycol_18 domain with two chitin-binding and chitin-catalyzing domains, and a C-terminal proline, glycine, serine, threonine (PGST)-rich domain. In addition, EaChi2 had a chitin-binding peritrophin-A domain at the end of the C-terminus with 5 cysteine residues possibly contributing two intradomain disulfide bonds. Multiple sequence alignment of the catalytic domain centers of glycol_18 domain displayed highly conserved chitin-binding and chitin-catalyzing domains in which three essential amino acid residues (D, D, E) for catalyzing activity are well conserved except EaChi4. The critical glutamic acid (E) residue was substituted for glutamine (Q) in EaChi4 indicating that it is devoid of catalytic activity. To our knowledge, this is the first report providing direct evidence that multiple earthworm chitinases are bacteria-responsive, strongly suggesting that earthworm chitinases are inducible humoral effectors in earthworm innate immunity. In addition, our results possibly suggest that earthworm EaChi4 may function as a pattern recognition molecule modulating the downstream immune pathway.

Benzoxazepine-Derived Selective, Orally Bioavailable Inhibitor of Human Acidic Mammalian Chitinase.

Human acidic mammalian chitinase (hAMCase) is one of two true chitinases in humans, the function of which remains elusive. In addition to the defense against highly antigenic chitin and chitin-containing pathogens in the gastric and intestinal contents, AMCase has been implicated in asthma, allergic inflammation, and ocular pathologies. Potent and selective small-molecule inhibitors of this enzyme have not been identified to date. Here we describe structural modifications of compound OAT-177, a previously developed inhibitor of mouse AMCase, leading to OAT-1441, which displays high activity and selectivity toward hAMCase. Significantly reduced off-target activity toward the human ether-à-go-go-related gene (hERG) and a good pharmacokinetic profile make OAT-1441 a potential candidate for further preclinical development as well as a useful tool compound to study the physiological role of hAMCase.

Comprehensive Analysis of the Chitinase Gene Family in Cucumber (Cucumis sativus L.): From Gene Identification and Evolution to Expression in Response to Fusarium oxysporum.

Chitinases, a subgroup of pathogenesis-related proteins, are responsible for catalyzing the hydrolysis of chitin. Accumulating reports indicate that chitinases play a key role in plant defense against chitin-containing pathogens and are therefore good targets for defense response studies. Here, we undertook an integrated bioinformatic and expression analysis of the cucumber chitinases gene family to identify its role in defense against Fusarium oxysporum f. sp. cucumerinum. A total of 28 putative chitinase genes were identified in the cucumber genome and classified into five classes based on their conserved catalytic and binding domains. The expansion of the chitinase gene family was due mainly to tandem duplication events. The expression pattern of chitinase genes was organ-specific and 14 genes were differentially expressed in response to F. oxysporum challenge of fusarium wilt-susceptible and resistant lines. Furthermore, a class I chitinase, CsChi23, was constitutively expressed at high levels in the resistant line and may play a crucial role in building a basal defense and activating a rapid immune response against F. oxysporum. Whole-genome re-sequencing of both lines provided clues for the diverse expression patterns observed. Collectively, these results provide useful genetic resource and offer insights into the role of chitinases in cucumber-F. oxysporum interaction.

Measurement of Chitinase Activity in Biological Samples.

Chitinases are the enzymes that cleave chitin. Even in the absence of chitin, mammalians have significant amounts of chitinases present in the body including at baseline. The precise role of chitinase is not known, however it was believed to play important role in digestion and host defense against chitin-containing food and pathogens, respectively. Recent work, including ours, has shown an important role of chitinase and chitinase-like proteins in host immunity and allergic diseases. Importantly, chitinase activities serve as important biomarkers of disease severity in a wide-range of diseases including type 2 inflammatory diseases such as asthma and pulmonary fibrosis. Similarly, patients with genetic disorders like Gaucher disease have significantly elevated chitinase levels, which not only correlate with disease severity but also serve as a reliable biomarker for therapeutic effectiveness. The protocol outlined here describes a simple, quick, and straightforward way to measure chitinase activity in BAL or serum samples of mice and can be widely adapted to human subjects and other model organisms due to the highly conserved nature of the enzymes.

The Genetic Polymorphisms of 24 Base Pair Duplication and Point G102S of Human Chitotriosidase to Bancroftian Filariasis at the Thai⁻Myanmar Border.

Lymphatic filariasis, caused by lymphatic filarial parasites, Wuchereria bancrofti, and Brugia malayi, causes significant morbidity and disability to 120 million people in the tropics and subtropics. Chitin has an important role for embryogenesis in adult worms and is a component of microfilaria sheath. Human chitotriosidase (CHIT1) is a chitin-degrading enzyme which provides a protective role against chitin-containing pathogens. Here, we determined the association of CHIT1 polymorphisms with susceptibility to bancroftian filariasis (BF) in 88 individuals at the Thai–Myanmar border. Two common polymorphisms of CHIT1, contributing inactive CHIT protein, including 24 base pair (24 bp) duplication in exon 10, and p. G102S in exon 4 were genotyped by allele-specific Polymerase Chain Reaction (PCR) and PCR sequencing, respectively. Unexpectedly, genotype frequencies of 24 bp duplication insertion homozygous (INS/INS) were significantly higher in endemic normal (EN) (40.0%) than BF patients (31.4%). In contrast, genotype frequencies of p. G102S homozygous (A/A) in BF patients (21.6%) was higher than in EN (19.0%) without statistical difference. Mutant allele frequencies of 24 bp duplication were 0.6125 (98/160) and p. G102S were 0.392 (69/176). Genotype and allele frequencies of CHIT1, 24 bp duplication, and p. G102S, showed no association with BF patients.

Human Chitinases: Structure, Function, and Inhibitor Discovery.

Chitinases are glycosyl hydrolases that hydrolyze the β-(1-4)-linkage of N-acetyl-D-glucosamine units present in chitin polymers. Chitinases are widely distributed enzymes and are present in a wide range of organisms including insects, plants, bacteria, fungi, and mammals. These enzymes play key roles in immunity, nutrition, pathogenicity, and arthropod molting. Humans express two chitinases, chitotriosidase 1 (CHIT1) and acid mammalian chitinase (AMCase) along with several chitinase-like proteins (CLPs). Human chitinases are reported to play a protective role against chitin-containing pathogens through their capability to degrade chitin present in the cell wall of pathogens. Now, human chitinases are gaining attention as the key players in innate immune response. Although the exact mechanism of their role in immune response is not known, studies in recent years begin to relate chitin recognition and degradation with the activation of signaling pathways involved in inflammation. The roles of both CHIT1 and AMCase in the development of various diseases have been revealed and several classes of inhibitors have been developed. However, a clear understanding could not be established due to complexities in the design of the right experiment for studying the role of human chitinase in various diseases. In this chapter, we will first outline the structural features of CHIT1 and AMcase. We will then review the progress in understanding the role of human chitinases in the development of various diseases. Finally, we will summarize the inhibitor discovery efforts targeting both CHIT1 and AMCase.

Phagocytosis-mediated M1 activation by chitin but not by chitosan.

Chitin particles have been used to understand host response to chitin-containing pathogens and allergens and are known to induce a wide range of polarized macrophage activations, depending, at least in part, on particle size. Nonphagocytosable particles larger than a macrophage induce tissue repair M2 activation. In contrast, phagocytosable chitin microparticles (CMPs, 1-10 μm diameters) induce M1 macrophages that kill intracellular microbes and damage tissues. However, chitosan (deacetylated) microparticles (de-CMPs, 1-10 µm) induce poor M1 activation. Toll-like receptor 2 (TLR2) and associated coreceptors in macrophages appear to be required for the M1 activation. To understand the exact mechanism of phagocytosis-mediated M1 activation by chitin, we isolated macrophage proteins that bind to CMPs during early phagocytosis and determined that TLR1, TLR2, CD14, late endosomal/lysosomal adaptor MAPK and mechanistic target of rapamycin activator 1 (LAMTOR1), Lck/Yes novel tyrosine kinase (Lyn), and β-actin formed phagosomal CMP-TLR2 clusters. These proteins were also detected in TLR2 phagosomal clusters in macrophages phagocytosing de-CMPs, but at relatively lower levels than in the CMP-TLR2 clusters. Importantly, CMP-TLR2 clusters further recruited myeloid differentiation primary response gene 88 (MyD88) and Toll-IL-1 receptor-containing adaptor protein (TIRAP) and phosphorylated Lyn, whereas neither the adaptors nor phosphorylated Lyn was detected in the de-CMP clusters. The results indicate that the acetyl group played an obligatory, phagocytosis-dependent role in the initiation of an integrated signal for TLR2-mediated M1 activation.

Genome-wide identification and expression analysis of chitinase gene family in Brassica rapa reveals its role in clubroot resistance

Chitinases, a category of pathogenesis-related proteins, are responsible for catalyzing the hydrolysis of chitin into the N-acetyl-d-glucosamine. Therefore, chitinases are believed to function as a guardian against chitin-containing pathogens. Here, we examined the role of the Brassica rapa chitinase family genes in clubroot disease. A total of 33 chitinase genes were identified and grouped into five classes based on their conserved domain. They were distributed unevenly across eight chromosomes in B. rapa, and 31 of them contained few introns (≤2). In addition, the expression of these genes was organ-specific, and 14 genes were expressed differentially in response to Plasmodiophora brassicae challenge of clubroot-susceptible (CS NIL) and resistant (CR NIL) lines. Furthermore, reduced pathogen DNA content and clubroot symptoms were observed in the CS NILs after their treatment with chitin oligosaccharides 24 h prior to inoculation with P. brassicae. The findings indicate that chitinases play a crucial role in pathogen resistance of the host plants. The results offer an insight into the role of chitinase in B. rapa–P. brassicae interaction.

Functional characterization of chitin-binding lectin from Solanum integrifolium containing anti-fungal and insecticidal activities

BackgroundAlong with the rapid development of glycomic tools, the study of lectin–carbohydrate interactions has expanded, opening the way for applications in the fields of analytic, diagnostic, and drug delivery. Chitin-binding lectins (CBLs) play roles in immune defense against chitin-containing pathogens. CBLs from species of the Solanaceae family, such as tomato, potato and jimsonweed, display different binding specificities to sugar chains containing poly-N-acetyllactosamine.ResultsIn this report, CBLs from Solanum integrifolium were isolated by ion exchange chromatography. The fractions showed hemagglutination activity (HA). The recombinant CBL in the 293F cell culture supernatant was able to inhibit the growth of Rhizoctonia solani and Colletotrichum gloeosporioide. Furthermore, the carbohydrate-binding property of CBLs was confirmed with the inhibition of HA. Binding of CBL to Spodoptera frugiperda (sf21) insect cells can partly be inhibited by N-Acetylglucosamine (GlcNAc), which is related to decrease mitochondrial membrane potential of sf21 cells.ConclusionsThe results showed that CBL exhibited antifungal properties and inhibited insect cell growth, which is directly correlated to the lectin-carbohydrate interaction. Further identification and characterization of CBLs will help to broaden their scope of application in plant defense and in biomedical applications.

Adiponectin inhibits eosinophil recruitment and is protective in invasive aspergillosis

Abstract Aspergillus fumigatus is a fungal pathogen associated with allergy, asthma, and invasive infection in susceptible individuals. Our previous results suggested that eosinophils may be a novel therapeutic target in invasive aspergillosis (IA) patients that do not respond to conventional pharmacotherapy. Eosinophil recruitment is induced by inhalation of the fungal cell wall polysaccharide chitin. However, the mechanisms of immune regulation that drive this recruitment are not well understood. We observed decreased levels of the metabolic-regulating cytokine adiponectin in the lungs and serum of mice after chitin aspiration, suggesting that adiponectin may be involved in the regulation of type 2 lung immune responses. Co-aspiration of adiponectin with chitin significantly reduced eosinophil accumulation in the lungs, and airway eosinophil recruitment in adiponectin-deficient mice was increased with chitin aspiration. In addition, chitin aspiration resulted in decreased surface expression of the adiponectin receptor AdipoR1 on eosinophils. Most significantly, airway accumulation of eosinophils, disease severity, and fungal burden were increased in adiponectin knockout mice with IA. This may be due to a direct inhibition of eosinophil recruitment, as recombinant adiponectin inhibited chemotaxis of bone marrow-derived eosinophils in vitro. These results suggest that adiponectin inhibits potentially detrimental lung eosinophil recruitment in response to chitin-containing pathogens. Increased understanding of the relationship between metabolic and immune regulation may ultimately result in novel therapies for obesity and allergic and infectious disease.

Human Chitotriosidase Is an Endo-Processive Enzyme.

Human chitotriosidase (HCHT) is involved in immune response to chitin-containing pathogens in humans. The enzyme is able to degrade chitooligosaccharides as well as crystalline chitin. The catalytic domain of HCHT is connected to the carbohydrate binding module (CBM) through a flexible hinge region. In humans, two active isoforms of HCHT are found–the full length enzyme and its truncated version lacking CBM and the hinge region. The active site architecture of HCHT is reminiscent to that of the reducing-end exo-acting processive chitinase ChiA from bacterium Serratia marcescens (SmChiA). However, the presence of flexible hinge region and occurrence of two active isoforms are reminiscent to that of non-processive endo-chitinase from S. marcescens, SmChiC. Although the studies on soluble chitin derivatives suggest the endo-character of HCHT, the mode of action of the enzyme on crystalline chitin is not known. Here, we made a thorough characterization of HCHT in terms of the mode of action, processivity, binding, and rate constants for the catalysis and dissociation using α-chitin as substrate. HCHT efficiently released the end-label from reducing-end labelled chitin and had also high probability (95%) of endo-mode initiation of processive run. These results qualify HCHT as an endo-processive enzyme. Processivity and the rate constant of dissociation of HCHT were found to be in-between those, characteristic to processive exo-enzymes, like SmChiA and randomly acting non-processive endo-enzymes, like SmChiC. Apart from increasing the affinity for chitin, CBM had no major effect on kinetic properties of HCHT.

Biotechnological potential of cold-adapted enzymes

Chitinases, the chitin-degrading enzymes, have been shown to play important role in defense against the chitin-containing fungal pathogens. In this study, we identified 48 chitinase-coding genes from the woody model plant Populus trichocarpa. Based on phylogenetic analysis, the Populus chitinases were classified into seven groups. Different gene structures and protein domain architectures were found among the seven Populus chitinase groups. Selection pressure analysis indicated that all the seven groups are under purifying selection. Phylogenetic analysis combined with chromosome location analysis showed that Populus chitinase gene family mainly expanded through tandem duplication. The Populus chitinase gene family underwent marked expression divergence and is inducibly expressed in response to treatments, such as chitosan, chitin, salicylic acid and methyl jasmonate. Protein enzymatic activity analysis showed that Populus chitinases had activity towards both chitin and chitosan. By integrating sequence characteristic, phylogenetic, selection pressure, gene expression and protein activity analysis, this study shed light on the evolution and function of chitinase family in poplar.

A novel chitinase from the earthworm, Eisenia andrei

ABSTRACTA novel chitinase gene, EaChi, and its expression pattern from the earthworm, E. andrei are demonstrated. Based on a deduced amino acid sequence, in EaChi, two specific domains for GH family 18 are well conserved with two essential amino acid residues for enzyme activity. The phylogenetic analysis shows that earthworm chitinase, EaChi, is evolutionarily close to other lophotrochozoan chitinases. The expression pattern analysis of EaChi indicates that the major expression is localized at intestinal epithelium and epidermis, possibly suggesting that the prime functions of the chitinase activity could be related to not only digestive process but also self-defending immunity as a biochemical barrier to protect the invasion of chitin-containing pathogens, including fungi, nematodes and protozoa.

Role of AMCase in the Allergic and Non Allergic Ocular Pathologies

Chitin is abundant in the structural coatings of fungi, insects, and parasitic nematodes, but it is not produced in mammals. The host defense against chitin-containing pathogens include production of chitinases. An acidic mammalian chitinase (AMCase) is produced in human epithelial cells of lower airways and conjunctiva via a Th2-specific, IL-13-dependent pathway and seems to be associated to asthma and allergic ocular pathologies. The role of AMCase in allergic disease is only at beginning and many issues open new possibilities for its control using specific inhibitors of AMCase activity or modulating its expression. In patients with vernal keratoconjunctivitis (VKC) and with season allergic conjunctivitis (SAC) the level of AMCase activity in the tears was found significantly elevated when compare to healthy controls and the highest levels were found in VKC. When RNA was extracted by conjunctival epithelial cells of these patients, the Real Time PCR measurement confirmed that the mRNA expression correlates with tear AMCase activity and the expression was significantly higher in VKC and SAC respectively. Also Receiver Operating Characteristic (ROC) analysis demonstrated that the sensitivity and specificity of AMCase measurement were 100% respectively, addressing the use of AMCase assay in the biochemical diagnosis of VKC and SAC. Recent studies in rabbit, where a reactive uveitis was induced by LPS injection in eye’s anterior chamber, confirmed that increased AMCAse activity was measurable in tears and that epithelial cells of conjunctiva express specific mRNA. A well as it was previously demonstrated in experimental model of mouse asthma, the inflammatory reaction induced by LPS was controlled by the chitinase inhibitor and steroid, instilled at 3 hr interval in conjunctival sacs. In dry eye, another pathologies where the role of innate immunity is sustained by AMCase secretion, an increased AMCase activity was documented and the specific mRNA expressed by epithelial conjunctival cells. In this pathology the eye inflammation can be ascribed to a common mechanism mediated by AMCase, via a Th 2 specific, IL-13 dependent way. In synthesis AMCase may be considered an important mediator in the pathogenesis of Th2 inflammation eye’s diseases, suggesting its potential diagnostic and therapeutic utility.

Presence of chitinase enzymes in crocodilians

Chitin is an abundant bio-polymer present as a structural component of many organisms such as arthropods, nematodes, mollusks, insects, and fungi, among others. Chitinolytic enzymes are synthesized for organisms to defend themselves against chitin-containing pathogens. Chitotriosidase (CHT) is a chitinase enzyme and one of the main proteins secreted by activated macrophages. It plays an important role in mechanisms of immunity by hydrolyzing chitin, thus protecting against chitin-containing pathogens. In this study, CHT was detected in Caiman latirostris plasma, and characterized under laboratory controlled conditions of temperature, reaction time, plasma concentration, pH and salinity. The results complement other immunological studies performed in caimans and demonstrate that they possess an efficient and well-developed immune system that resists the attack of some pathogens. Based on the current knowledge of the properties and homologies of CHT, it would be highly valuable to evaluate its possible therapeutic application in the veterinary clinical setting.