Termite Extracts Research Articles

CRUDE CELLULASES FROM SUBTERRANEAN TERMITE EXTRACT (Macrotermes gilvus Hagen) FOR SACCHARIFICATION OF LEAF LITTER AND BIOSOFTENING PROCESS IN SKELETON LEAF CRAFTS MAKING

Recently, leaf litter has been used as compost or piled up and burned openly in most regions of Indonesia. This litter has the potential for its lignocellulosic components as the source of sugars. Also, the leave skeleton structure is promising for creative industrial purposes. Besides, termites (Macrotermes gilvus Hagen) can digest lignocellulosic biomass containing high fiber. In this research, termite extract contained lignocellulases and was used in the leaf litter pretreatment process for sugar preparation and leaf litter biosoftening. This study aimed to extract termite enzymes, identify, evaluate the activity of crude enzymes, and characterize the hydrolytic ability of enzymes to degrade lignocellulosic materials in three different leaf litters, i.e., Polyalthia longifolia (PL), Pterocarpus indicus (PI), and Filicium decipiens (FD). The leaf litter biosoftening process was performed in pulp removal for Annona muricata (AM) skeleton leaf crafts making. The results showed that M. gilvus Hagen extract exhibited high activity of endo-β-D-1,4-glucanase (CMCase), moderate activity of avicelase, β-D-1,4-mannanase, β-D-1,4-xylanase, and low β-D-1,4-glucosidase. The optimum pH of CMCase, β-D-1,4-mannanase, and β-D-1,4-glucosidase were 6.5, 5.0, and 6.0, respectively, while the optimum temperatures were 40 °C, 50 °C, and 55 °C, respectively. Cellulases from termite extracts were suitable pretreatment of leaf litter biomass, as evidenced by saccharification activity, with the highest saccharification activity in FD leaf litter. Enzymes were also successfully employed to help remove AM leaf pulp at the volume of 100 mL for 14 days, with the removal efficiency at 98.7%.

Prospect of propolis from stingless bee, Heterotrigona itama as biological control of the subterranean termite, Coptotermes curvignathus

This study applies a propolis from a stingless bee, Heterotrigona itama as an alternative to control the infestation of a subterranean termite, Coptotermes curvignathus. The objective of this study was to assess the antitermitic activity of stingless bee’s propolis as termiticide based on its repellency activity, contact toxicity effect as well as its ability to act as cellulase inhibitor. The bioactive components of the propolis in terms of the phenolics and flavonoids content were also determined. It was observed that the propolis is a termite’s repellent with a preference index of -0.73. Upon contact, it can kill 50% of the termite’s population within 1.5 hours with lethal concentration of about 16% (w/v). It’s extract also can inhibit cellulase activity of termites. Diameter of the clear zone on the CMC agar was found to be significantly reduced from 3.1 cm to 2.4 cm when 30% (w/v) propolis’s extract was added into the termite extract at the ratio of 1 to 4 (termite: propolis extract). In conclusion, from the results obtained, propolis from Heterotrigona itama has high potential to be used as an environmentally safe alternative to chemical termiticide.

Cloning, Expression, and Immunological Characterization of Formosan Subterranean Termite (Blattodea: Rhinotermitidae) Arginine Kinase.

Several parts of the world regularly consume termites. Arthropod arginine kinase proteins often cross-react with human immunoblobulin E (IgE) antibodies and they are considered pan-allergens. The Formosan subterranean termite Coptotermes formosanus (C. formosanus (Shiraki) [Isoptera: Rhinotermitidae]), along with cockroaches, belong to the order Blattodea and they are common household pests in tropical and subtropical parts of the world. An sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) band migrating at approximately 37 kDa in C. formosanus termite extracts cross-reacted with IgE from five cockroach allergic patient samples by immunoblot. Liquid chromatography-mass spectrometry analysis of gel slices from the corresponding region of a gel indicated several peptides from the excised region were identical to the American cockroach arginine kinase allergen, Per a 9. The sequence of the full-length C. formosanus arginine kinase gene indicates the protein it encodes is 96% identical to American cockroach Per a 9, 94% identical to German cockroach Bla g 9, and 82-84% identical to shrimp arginine kinase proteins Pen m 2, Lit v 2, and Cra c 2. Full-length C. formosanus arginine kinase was fused to a glutathione S-transferase tag and recombinantly expressed and purified from Escherichia coli by affinity chromatography. The recombinant protein was recognized by IgE from 11 of 12 cockroach or shrimp allergic samples, but did not cross-react with dust mite allergic or peanut/tree nut allergic samples. The results of this study indicate the C. formosanus arginine kinase cross-reacts with cockroach and shrimp allergic IgE, and if consumed would likely act as an allergen.

Ethnomedicinal survey and in vitro confirmation of anti-inflammatory and antispasmodic properties of the termite strain Macrotermes bellicosus used in traditional medicine in the Republic of Benin.

Ethnopharmacological relevanceInsects and insect-derived products play a vital role in traditional medicine in many parts of the world since ancient times. Among these insects, fungus-growing termites like Macrotermes bellicosus (M. bellicosus) are widely used in nutrition and traditional medicine in various societies of sub-Saharan Africa. Aim of the studyAim of the present study was to explore the traditional applications of M. bellicosus and subsequently investigate the anti-inflammatory and spasmolytic activity of samples collected in Benin. Material and methodsAn ethnomedicinal survey with thirty active healers in Benin was conducted and the anti-inflammatory activity of an ethanolic extract of M. bellicosus was investigated. Thus, LPS-induced TNFα release from differentiated human macrophages (THP-1) and IL-8 release from cytokine (IL-1β/TNFα/IFNγ)-challenged human intestinal epithelial cells (Caco-2) was measured by enzyme-linked immunosorbent assay. Furthermore, the influence of M. bellicosus extract on basal tone and induced contractions in isolated rat small intestinal preparations was determined to examine the influence on intestinal motility. ResultsThe survey of 30 active healers demonstrated that M. bellicosus and its products (termites’ mound and fungus comb) are used in Benin for therapeutic purposes mainly to treat infectious and inflammatory diseases including digestive disorders, snake bites and diarrhea. It was found that M. bellicosus extract inhibited both LPS-induced TNFα release from human macrophages and cytokine-induced IL-8 release from intestinal epithelial cells comparable to budesonide. In addition, isometric contraction measurement with isolated rat small intestinal preparations demonstrated a mild spasmolytic effect of the termite extract in higher concentrations with a suppression of induced contractions and relaxation of basal tone. ConclusionM. bellicosus which is used in traditional medicine in Benin to treat infectious and inflammatory diseases showed anti-inflammatory activity by inhibiting pro-inflammatory cytokine release and a moderate influence on intestinal motility.

Cross-reaction between Formosan termite (Coptotermes formosanus) proteins and cockroach allergens.

Cockroach allergens can lead to serious allergy and asthma symptoms. Termites are evolutionarily related to cockroaches, cohabitate in human dwellings, and represent an increasing pest problem in the United States. The Formosan subterranean termite (Coptotermes formosanus) is one of the most common species in the southern United States. Several assays were used to determine if C. formosanus termite proteins cross-react with cockroach allergens. Expressed sequence tag and genomic sequencing results were searched for homology to cockroach allergens using BLAST 2.2.21 software. Whole termite extracts were analyzed by mass-spectrometry, immunoassay with IgG and scFv antibodies to cockroach allergens, and human IgE from serum samples of cockroach allergic patients. Expressed sequence tag and genomic sequencing results indicate greater than 60% similarity between predicted termite proteins and German and American cockroach allergens, including Bla g 2/Per a 2, Bla g 3/Per a 3, Bla g 5, Bla g 6/Per a 6, Bla g 7/Per a 7, Bla g 8, Per a 9, and Per a 10. Peptides from whole termite extract were matched to those of the tropomyosin (Bla g 7), arginine kinase (Per a 9), and myosin (Bla g 8) cockroach allergens by mass-spectrometry. Immunoblot and ELISA testing revealed cross-reaction between several proteins with IgG and IgE antibodies to cockroach allergens. Several termite proteins, including the hemocyanin and tropomyosin orthologs of Blag 3 and Bla g 7, were shown to crossreact with cockroach allergens. This work presents support for the hypothesis that termite proteins may act as allergens and the findings could be applied to future allergen characterization, epitope analysis, and clinical studies.

IDENTIFICATION AND EVALUATION OF FIBER HYDROLYTIC ENZYMES IN THE EXTRACT OF TERMITES (Glyptotermes montanus) FOR POULTRY FEED APPLICATION

Foot rot disease of black pepper caused by Phytophthora capsici had been reported in Batangas and Laguna, Philippines. The plant was recovered following the application of crop residue (organic substrate) and intercropping with other crops. This study was aimed to isolate, identify, and determine the soil mycoflora from the rhizosphere of black pepper grown on various cropping patterns in Batangas and Laguna. Antagonistic activity of mycoflora isolates was tested against P. capsici using dual culture technique. The result showed that 149 colonies of soil mycoflora isolated were belonging to 14 genera; three of them, i.e. Penicillium, Paecilomyces and Aspergillus, were the most dominant. All of the mycoflora isolates were able to inhibit the growth of the pathogen. Eighteen of them were the most promising antagonists, based on their inhibition growth of more than 60%. It is suggested that antagonistic mechanism of Mucor isolate (1001), Trichoderma (125, 170, 171, 179, 180, 181), Gliocladium (109), Cunninghamella (165, 168), Mortierella (177), and Aspergillus (106) was space competitor (competition for nutrient) since they rapidly overgrew the pathogen. Aspergillus (67, 79, 81, 83, 108, and 202) isolates inhibited the pathogen apparently by producing antibiotic, whereas Trichoderma (125, 170, 171, 179, 180, and 181) isolates were able to penetrate the hyphae of the pathogen. The organic matter percentage in the soil was significantly correlated with the number of antagonistic mycoflora in rhizosphere (R2 = 0.1094), but the cropping pattern wPoultry are not able to digest fiber in the diet. Hydrolytic enzymes including cellulases and hemicellulases have been used as poultry feed supplement. Termites (Glyptotermes montanus) have the ability to digest wood that contains high fiber. The purpose of this experiment was to identify the cellulase and hemicellulase of termite extract. The hydrolytic (saccharification) activity of the termite extract on feedstuffs was then evaluated. It contained high endo-β-D-1,4-glucanase (CMCase) activity, but the activities of avicelase, β-D-1,4-mannanase, β- D-1,4-xylanase, and β-D-1,4-glucosidase were very low. The activities of the enzymes were higher in the fresh extract than those extracted after drying at 40oC with blower oven. CMCase (as cellulase), β-D-1,4-mannanase (as hemicellulase), and β-D- 1,4-glucosidase (as glycosidase) were reevaluated further to determine the optimum pH and temperatures for maximum activities. The optimum pH for CMCase, β-D-1,4-mannanase, and β-D-1,4-glucosidase were 6.2, 5.0, and 5.8 respectively, while the optimum temperatures were 45-50oC, 50-55oC, and 42-45oC, respectively. The enzyme mixture or cocktail was more appropriate in digesting feedstuffs with high lignocellulose (fiber) such as rice bran and pollard than feedstuffs with more soluble starch such as soybean and corn meals. The extracted enzyme could be immobilized with pollard, but CMCase recovery was low (28.6%), while β-D-1,4-mannanase and β-D-1,4-glucosidase recoveries were 89.2% and 272.9%, respectively. Termite extract contained enzyme cocktails of lignocellulases that potentially be used as feed supplement. However, its use is limited by its low activity.as negatively correlated. This study suggests that organic matter increased antagonistic mycoflora in black pepper rhizosphere, which will reduce severity of the disease.

IDENTIFICATION AND EVALUATION OF FIBER HYDROLYTIC ENZYMES IN THE EXTRACT OF TERMITES (Glyptotermes montanus) FOR POULTRY FEED APPLICATION

Foot rot disease of black pepper caused by Phytophthora capsici had been reported in Batangas and Laguna, Philippines. The plant was recovered following the application of crop residue (organic substrate) and intercropping with other crops. This study was aimed to isolate, identify, and determine the soil mycoflora from the rhizosphere of black pepper grown on various cropping patterns in Batangas and Laguna. Antagonistic activity of mycoflora isolates was tested against P. capsici using dual culture technique. The result showed that 149 colonies of soil mycoflora isolated were belonging to 14 genera; three of them, i.e. Penicillium, Paecilomyces and Aspergillus, were the most dominant. All of the mycoflora isolates were able to inhibit the growth of the pathogen. Eighteen of them were the most promising antagonists, based on their inhibition growth of more than 60%. It is suggested that antagonistic mechanism of Mucor isolate (1001), Trichoderma (125, 170, 171, 179, 180, 181), Gliocladium (109), Cunninghamella (165, 168), Mortierella (177), and Aspergillus (106) was space competitor (competition for nutrient) since they rapidly overgrew the pathogen. Aspergillus (67, 79, 81, 83, 108, and 202) isolates inhibited the pathogen apparently by producing antibiotic, whereas Trichoderma (125, 170, 171, 179, 180, and 181) isolates were able to penetrate the hyphae of the pathogen. The organic matter percentage in the soil was significantly correlated with the number of antagonistic mycoflora in rhizosphere (R2 = 0.1094), but the cropping pattern wPoultry are not able to digest fiber in the diet. Hydrolytic enzymes including cellulases and hemicellulases have been used as poultry feed supplement. Termites (Glyptotermes montanus) have the ability to digest wood that contains high fiber. The purpose of this experiment was to identify the cellulase and hemicellulase of termite extract. The hydrolytic (saccharification) activity of the termite extract on feedstuffs was then evaluated. It contained high endo-β-D-1,4-glucanase (CMCase) activity, but the activities of avicelase, β-D-1,4-mannanase, β- D-1,4-xylanase, and β-D-1,4-glucosidase were very low. The activities of the enzymes were higher in the fresh extract than those extracted after drying at 40oC with blower oven. CMCase (as cellulase), β-D-1,4-mannanase (as hemicellulase), and β-D- 1,4-glucosidase (as glycosidase) were reevaluated further to determine the optimum pH and temperatures for maximum activities. The optimum pH for CMCase, β-D-1,4-mannanase, and β-D-1,4-glucosidase were 6.2, 5.0, and 5.8 respectively, while the optimum temperatures were 45-50oC, 50-55oC, and 42-45oC, respectively. The enzyme mixture or cocktail was more appropriate in digesting feedstuffs with high lignocellulose (fiber) such as rice bran and pollard than feedstuffs with more soluble starch such as soybean and corn meals. The extracted enzyme could be immobilized with pollard, but CMCase recovery was low (28.6%), while β-D-1,4-mannanase and β-D-1,4-glucosidase recoveries were 89.2% and 272.9%, respectively. Termite extract contained enzyme cocktails of lignocellulases that potentially be used as feed supplement. However, its use is limited by its low activity.as negatively correlated. This study suggests that organic matter increased antagonistic mycoflora in black pepper rhizosphere, which will reduce severity of the disease.

Functional characterization and target discovery of glycoside hydrolases from the digestome of the lower termite Coptotermes gestroi.

BackgroundLignocellulosic materials have been moved towards the forefront of the biofuel industry as a sustainable resource. However, saccharification and the production of bioproducts derived from plant cell wall biomass are complex and lengthy processes. The understanding of termite gut biology and feeding strategies may improve the current state of biomass conversion technology and bioproduct production.ResultsThe study herein shows comprehensive functional characterization of crude body extracts from Coptotermes gestroi along with global proteomic analysis of the termite's digestome, targeting the identification of glycoside hydrolases and accessory proteins responsible for plant biomass conversion. The crude protein extract from C. gestroi was enzymatically efficient over a broad pH range on a series of natural polysaccharides, formed by glucose-, xylose-, mannan- and/or arabinose-containing polymers, linked by various types of glycosidic bonds, as well as ramification types. Our proteomic approach successfully identified a large number of relevant polypeptides in the C. gestroi digestome. A total of 55 different proteins were identified and classified into 29 CAZy families. Based on the total number of peptides identified, the majority of components found in the C. gestroi digestome were cellulose-degrading enzymes. Xylanolytic enzymes, mannan- hydrolytic enzymes, pectinases and starch-degrading and debranching enzymes were also identified. Our strategy enabled validation of liquid chromatography with tandem mass spectrometry recognized proteins, by enzymatic functional assays and by following the degradation products of specific 8-amino-1,3,6-pyrenetrisulfonic acid labeled oligosaccharides through capillary zone electrophoresis.ConclusionsHere we describe the first global study on the enzymatic repertoire involved in plant polysaccharide degradation by the lower termite C. gestroi. The biochemical characterization of whole body termite extracts evidenced their ability to cleave all types of glycosidic bonds present in plant polysaccharides. The comprehensive proteomic analysis, revealed a complete collection of hydrolytic enzymes including cellulases (GH1, GH3, GH5, GH7, GH9 and CBM 6), hemicellulases (GH2, GH10, GH11, GH16, GH43 and CBM 27) and pectinases (GH28 and GH29).

Elucidating the Redox Cycle of Environmental Phosphorus Using Ion Chromatography

Historically, it was assumed that reactive, inorganic phosphorus present in pristine environments was solely in the form of orthophosphate. However, this assumption contradicts theories of biogenesis and the observed metabolic behavior of select microorganisms. This paper discusses the role of ion chromatography (IC) in elucidating the oxidation-reduction cycle of environmental phosphorus. These methods employ suppressed-IC, coupled with tandem conductivity and electrospray mass spectrometry detectors to identify and quantify phosphorus oxyanions in natural water, synthetic cosmochemical, and biological samples. These techniques have been used to detect phosphite and orthophosphate in geothermal hot springs. Hypophosphite, phosphite, and orthophosphate have been detected in synthetic schreibersite corrosion samples, and termite extract supernatant. Synthetic schreibersite corrosion samples were also analyzed for two poly-phosphorus compounds, hypophosphate and pyrophosphate, and results show these samples did not contain concentrations above the 1.3 and 2.0 μM respective 3σ limit of detection. These methods are readily adaptable to a variety of matrices, and contribute to the elucidation of the oxidation-reduction cycle of phosphorus oxyanions in the environment. In contrast to most studies, these techniques have been used to show that phosphorus actively participates in redox processes in both the biological and geological world.

Purification and Molecular Cloning of Xylanases from the Wood-Feeding Termite,Coptotermes formosanusShiraki

Coptotermes formosanus is one of the most destructive termites in the southern part of Japan as well as in the United States. Hemicellulose is a noncellulosic polysaccharide found in plant cell walls, and xylan is the major constituent of hemicellulose. Since hemicellulose prevents access of cellulolytic enzymes to cellulose, enzymatic hydrolysis of hemicellulose is beneficial for cellulose digestion. We purified three functional xylanases to homogeneity from C. formosanus for the first time. Elution profiles from the whole termite extract suggest that these three xylanases play major roles in xylan digestion in the gut of the termites. The corresponding cDNAs were successfully cloned based on the N-terminal amino acid sequences, encoding GHF11 xylanases. Reverse transcription-PCR using manipulated protozoan cells in the hindgut revealed that the corresponding genes were expressed in the symbiotic flagellate Holomastigotoides mirabile. These results suggest that the GHF11 xylanases that are produced by the symbiotic flagellates play a primary role in xylan degradation in C. formosanus.

Utilization of mixed cellulolytic microbes from termite extract, elephant faecal solution and buffalo ruminal fluid to increase in vitro digestibility of King Grass

Cellulose is a compound of plant cell walls which is difficult to be degraded because it composed of glucose monomers linked by β-(1.4)-bound. It will be hydrolysed by cellulase enzyme secreted by cellulolytic microbes. The effective digestion of cellulose needs high activity of cellulase enzyme. This research aims to increase in vitro king grass digestibility utilizing mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid. Twelve syringes contained gas test media were randomly divided into four treatments based on sources of microbe (SM), namely: S (SM: cattle ruminal fluid [S]), RGK (SM: mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid [RGK], with composition 1 : 1 : 1), S-RGK (SM: S + RGK, with composition 1:1), and TM (without given treatment microbe). Digestibility was measured using gas test method. Average of gas production treatment of S-RGK (70.2 + 0.6 ml) was higher and significantly different (P<0.01) compared to treatment of S (60.3 + 0.8 ml), RGK (40.8 + 2.3 ml), and TM (13.3 + 2.0 ml). Utilization of mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid (RGK) that combined with microbes of cattle ruminal fluid (S) could increase in vitro digestibility of king grass. Key Words : Cellulolytic Microbe, Termite Extract, Elephant Faecal, Buffalo Ruminal Fluid

Enzymes of acetate and glucose metabolism in termites

Extracts of tissues of the lower termites, Reticulitermes flavipes and Coptotermes lacteus, and the higher termite, Nasutitermes exitiosus, possess acetyl-CoA synthetase and all the enzymes of the tricarboxylic acid cycle and are thus able to oxidize acetate to CO 2. The specific activities of these enzymes in R. flavipes are sufficient to cope with the rate of acetogenesis by the gut microbiota. The presence of the malic enzyme and malate dehydrogenase, but not pyruvate carboxylase or phosphoenolpyruvate carboxykinase, indicates that they may be important as anaplerotic enzymes for the conversion of pyruvate to oxalacetate. An apparent absence of pyruvate dehydrogenase in all termites suggests that they do not convert pyruvate to acetyl-CoA, but rather convert acetate (transported from the hindgut) to this compound. All the enzymes of glycolysis were present in termite extracts. Thus any glucose absorbed from the midgut, and originating from hydrolysis of cellulose by salivary or midgut enzymes, can be metabolized by termites as an energy source.

Alteration of the gut microbiota and its effect on nitrogen metabolism in termites

When the spirochaetes in the gut of Nasutitermes exitiosus were killed with oxygen, the glutamate dehydrogenase activity of whole termite extracts declined by some 75%. Starvation, which caused a similar termite mortality rate to oxygen treatment, did not greatly affect this enzyme. Treatment of N. exitiosus with tetracycline to kill all the gut flora led to glutamate dehydrogenase activity falling to zero. Treatment of Coptotermes lacteus with oxygen to kill gut protozoa and spirochaetes resulted in the activity of glutamate dehydrogenase remaining low, whereas in starved and normal termites the activity of the enzyme increased about seven-fold. A similar result was observed when tetracycline was used to kill the gut flora and fauna. Aspartate aminotransferase and alanine aminotransferase activities (which appear to be of termite origin) were little affected by these treatments. The glutamate dehydrogenase was NAD +-dependent and was not found in the bacteria (excluding spirochaetes) colonising the hindgut. In all experiments there was a slight increase in the uric acid content of the termites in both control and treated insects. The free amino acid content of both species of termites was increased (about 50%) when they were starved, but was little affected when the gut flora and/or fauna were killed. Thus the gut flora and fauna do not appear to be involved in amino acid metabolism. When C. lacteus was kept under argon-oxygen mixtures for seven weeks the total nitrogen, enzyme activities and uric acid content were similar to those of termites kept under nitrogen-oxygen mixtures. These results indicate that in the short term termites do not need to fix nitrogen and also that nitrogen is well conserved.

Uric acid in wood-eating termites

Uric acid (UA) was present in termites as assessed by paper and thin layer chromatography, by U.V. spectroscopy, by reactivity with uricase, and by gas chromatography coupled to mass spectrometry. Specimens of six species ( Reticulitermes flavipes, R. virginicus, Coptotermes formosanus, Marginitermes hubbardi, Paraneotermes simplicicornis, and Cryptotermes cavifrons) contained UA in amounts accounting for between 1 and 45% of the termites' dry weight and for between 4 and 69% of the termites' total nitrogen. Almost all of the UA in R. flavipes was associated with fat body tissue. Faeces contained less than 0.2% of its weight as UA. During 15 months of captivity, UA in R. flavipes workers increased from 1.7% to 45.4% of the insects' dry weight at a linear rate of 2.7% per month. Elemental analyses of termites during this period indicated that synthesis of UA occurred, in part, at the expense of some endogenous nitrogen and carbon. Compounds tentatively identified as inosine and kynurenic acid were also present in termite extracts. Results indicate that termites can store large quantities of UA internally, but they do not void the purine in significant amounts despite the lack of detectable uricase activity in termite tissues.

An investigation of chitinase activity in cockroach and termite extracts

The conditions for an unequivoval demonstration of chitinase activity in insect extracts have been investigated. The breakdown of undenatured chitin by cockroach preparations (blood, digestive juice, cuticle brei, extract of cast skins, saliva) and by an extract of whole termites has been demonstrated. The pH optimum for cockroach digestive chitinase lies between 5·4 and 6·0 and the products of digestion of chitin with this preparation were N-acetylglucosamine (75 per cent), probably its dimer and trimer, and a trace of glucosamine. Only N-acetylglucosamine and a trace of glucosamine were produced by the action of other cockroach preparations. Deacetylase activity could not be detected in the cockroach preparations.