Single Nitrogen Source Research Articles

Study on characteristics of biocometabolic removal of omethoate by the Aspergillus spp .

A strain Aspergillus spp. F1 which could effectively metabolized omethoate was screened out in this study. F1 tended to form granula with diameter 4–5 mm after 5 days culture in shaker. The pH range from 4.5 to 6.5 was the suitable pH range for growth and metabolism of Aspergillus spp. F1. The maximum omethoate removal rate was about 3.0 mg/(h L), and the removal fraction of omethoate reached 90% after 8 days culture when initial concentrations of omethoate were not more than 2000 mg/L in medium. There was no obvious relativity between cell growth and cometabolism of omethoate. Starch was the best carbon source for omethoate removal and the result after 3 days reached 56.6% removal. F1 could use omethoate to metabolize as single nitrogen or phosphate source. The residual fragments in medium after treatment with the Aspergillus spp. F1 were determined by gas chromatography–mass spectrometry. The analysis results showed that only fragment o,o,s,-trimethyl phosphorothioate (TEP) containing phosphorus was available and the fragment containing nitrogen was consumed by F1 thoroughly in culture process. But no accumulations of TEP were observed in the omethoate bioremediation process. F1 could effectively remove omethoate in complex nutritional environment safely.

COMPLEX APPROACH TO THE PROBLEM OF PERSISTENT ORGANIC POLLUTANTS DEGRADATION IN WATER ENVIRONMENT

This paper deals with the physical-chemical and microbiologic treatment of persistent organic pollutants POPs (namely, benzothiazoles), and the limits of these methods applicability are considered. The new technical solutions improving their degradation efficiency were proposed, by using microbiological treatment and photo-catalytic one, with the UV-irradiation having the wavelength 180-320 nm in oxidative environment, containing hydrogen peroxide and iron (III) ions as catalysts. The kinetics and mechanism of benzothiazole (BT) molecules degradation was studied by gas-liquid and thin-layer chromatography, and intermediate products were identified. The role of redox processes, running under the influence of active radicals, is discussed. The screening of bacterial and fungal cultures made it possible to select certain strains, possessing different capacity of BTs utilization as the single source of nitrogen, carbon and energy. The study of the products of BTs microbial transformation products was carried out. The biodegradation of BT and OBT (hydroxybenzothiazole) by a few stems of fungi g. Penicillium was monitored by reverse phase HPLC method performed directly on culture media without purification. The most active biodestructors turned out to be Penicillium sp.24, Penicillium sp.77. The xenobiotics were biotransformed into hydroxylated derivatives.

Differentiation of Malassezia furfur and Malassezia sympodialis by glycine utilization

The genus Malassezia has been revised to include six lipophilic species and one nonlipophilic species. These Malassezia species have been investigated to differentiate their morphological and physiological characteristics. However, assimilation of amino acids as a nitrogen source by these species was not well elucidated. In the present study, isolates of Malassezia species were examined with a glycine medium (containing 7-266 mmol glycine, 7.4 mmol KH(2)PO(4), 4.1 mmol MgSO(4)7H(2)O, 29.6 mmol thiamine, 0.5% Tween-80 and 2% agar) and a modified Dixon glycine medium (0.6% peptone, 3.6% malt extract, 2% ox-bile, 1% Tween-40, 0.2% glycerol, 0.2% oleic acid, 7 mmol glycine and 2% agar). All M. furfur isolates developed on the glycine medium, assimilating glycine at concentrations of at least 7 mmol l(-1). However, the other six Malassezia species were unable to grow on the glycine medium. Also, many colonies of M. furfur grew rapidly, within 2-3 days on the modified Dixon glycine medium, although the other six species showed slow and poor development. From these results, it was suggested that M. furfur might be able to utilize glycine as a single nitrogen source, which the other Malassezia species could not. Therefore, glycine medium was recommended for the differentiation of M. furfur from other species of Malassezia.

Comparative growth of ectomycorrhizal basidiomycetes (Hebeloma spp.) on organic and inorganic nitrogen

In the largely organic soils in which ectomycorrhizas are commonly found, a preference for absorbing organic nitrogen over mineral forms is likely to be an advantage, especially where mineralisation rates are low. To determine rates of both independent and preferential growth of ectomycorrhizal basidiomycetes on organic and inorganic nitrogen, strains of Hebeloma were grown on nutrient agar media containing either NH4+ or glutamic acid as the sole source of nitrogen, on both single medium and split plate Petri dishes. Growth rates on the split plate Petri dishes, where the fungi had access to both nitrogen sources, were generally greater than on the single medium dishes. Growth on glutamic acid was at least equal to, and usually greater than, that on NH4+. In some cases growth on NH4+ alone appeared severely inhibited, a condition that was partially alleviated by access to glutamic acid on the split plates Petri dishes. This highlights a potential pitfall of single nitrogen source growth studies. The greater growth of most strains on glutamic acid suggests an adaptation to organic nitrogen utilisation in these strains. If this is so in soils with low mineralisation rates, direct uptake of amino acids by ectomycorrhizal plants could by-pass the bottle neck that requires mineral nitrogen to be made available for plant uptake.

Waste Reduction in Alkyl Pyridine Production: Solving the Root Problem Pays off

The production process for 5-ethyl-2-methyl-pyridine (MEP) is the largest single source of total organic carbon and total nitrogen in the feed of the capacity-limited wastewater treatment plant at LONZA's major production site. According to LONZA's integrated waste treatment concept, efforts were made to reduce the amount of critical components in the MEP process wastewater. Detailed analysis of causes and effects revealed the formation of small amounts of ethylamines in the reaction unit as the root cause not only for the ecological but also for the economical problems such as high catalyst consumption and reduced MEP yield. Separation of interfering ethylamines from the process in combination with recycling of catalyst and catalyst byproduct-containing process water can solve all problems.

Production of isomeric 9,10,13 (9,12,13)-trihydroxy-11 E (10 E )-octadecenoic acid from linoleic acid by Pseudomonas aeruginosa PR3

Trihydroxy unsaturated fatty acids with 18 carbons have been reported as plant self-defense substances. Their production in nature is rare and is found mainly in plant systems. Previously, we reported that a new bacterial isolate, Pseudomonas aeruginosa PR3, converted oleic acid and ricinoleic acid to 7,10-dihydroxy-8(E)-octadecenoic acid and 7,10,12-trihydroxy-8(E)-octadecenoic acid, respectively. Here we report that strain PR3 converted linoleic acid to two compounds: 9,10,13-trihydroxy-11(E)-octadecenoic acid (9,10,13-THOD) and 9,12,13-trihydroxy-10(E)-octadecenoic acid (9,12,13-THOD). Stereochemical analyses showed the presence of 16 different diastereomers — the maximum number possible. The optimum reaction temperature and pH for THOD production were 30°C and 7.0, respectively. The optimum linoleic acid concentration was 10 mg/ml. The most effective single carbon and nitrogen sources were glucose and sodium glutamate, respectively. However, when a mixture of yeast extract (0.05%), (NH4)2HPO4 (0.2%), and NH4NO3 (0.1%) was used as the nitrogen source, THOD production was higher by 8.3% than when sodium glutamate was the nitrogen source. Maximum production of total THOD with 44% conversion of substrate was achieved at 72 h of incubation, after which THOD production plateaued up to 240 h. THOD production and cell growth increased in parallel with glucose concentration up to 0.3%, after which cell growth reached its maximum and THOD production did not increase. These results suggested that THODs were not metabolized by strain PR3. This is the first report of microbial production of 9,10,13- and 9,12,13-THOD from linoleic acid. Journal of Industrial Microbiology & Biotechnology (2000) 25, 109–115.

Estimation of the impact of fertilisation rate on nitrate leaching in Finland using a mathematical simulation model

Agriculture comprises the largest single source of nitrogen (N) into watercourses in Finland. The emphasis of water protection policy is today on controlling the non-point nutrient losses from agriculture. In this study a mathematical simulation model was used as a management tool to estimate the changes in nitrate (NO 3 −-N) leaching resulting from changes in cultivation practices in Finnish agriculture caused by the Agri-Environmental Support Scheme as a part of the Common Agricultural Policy of the European Union. Detailed data were collected from about 400 farms by interviewing farmers in four study areas in different parts of the country. The potential impacts of these changes on nitrate losses were then assessed by a deterministic nitrogen leaching model and regional assessments were made by combining the results of model calculations with digital spatial data about soils, crops and fertilisation, using GIS-software. In general, the use of nitrogen fertilisers (inorganic fertilisers and manure) has decreased to meet the requirements of the Support Scheme. However, the estimated potential impacts on nitrate losses were rather small (3–14% in different study areas). The model results showed that to achieve the targeted national and international reductions in agricultural nitrate leaching, fertilisation and particularly manure spreading should be reduced and adjusted better to the actual nitrogen requirements of crops.

Effect of Mutations in Genes PH085 and PH04 on Proline Utilization in Yeast Saccharomyces cerevisiae

Yeast Saccharomyces cerevisiae can utilize proline as a single nitrogen source. We have found that the strains with mutations in gene PH085, a structural gene encoding cyclin-dependent phosphoprotein kinase, cannot grow on the proline-containing media. The ability to utilize proline is restored in strains of the pho85pho4genotype. We suggest that phosphoprotein kinase Pho85p is involved in either phosphorylation of a highly specific proline permease, Put4p, or in signaling proline concentration. The Pho4p protein that activates transcription of the PH05 gene, a structural gene of acid phosphatase, seems to participate in the negative regulation of the PUT1 and PUT2 genes encoding enzymes of proline catabolism, proline oxidase and Δ-pyrroline-5-carboxylate dehydrogenase. Thus, regulation of phosphorus and nitrogen metabolism have common elements.

Evaluation of sole nitrogen sources for shoot and leaf disc cultures of sugarbeet

Use of single nitrogen sources in nutrient media is essential to ascertaining the relative role and regulation of nitrogen assimilatory steps, and may help identify and understand highly productive media for micropropagation and adventitious shoot formation. Eight endogenous nitrogen-containing ions or compounds in sugarbeet (nitrate, ammonium, glutamine, glutamate, urea, proline, glycine betaine and choline) were examined for ability to serve as sole nitrogen source for shoot or leaf disc culture of sugarbeet (Beta vulgaris L.) model clone REL-1. The most productive concentrations of nitrate, ammonium, urea, and glutamine as sole nitrogen sources were moderately supportive of shoot multiplication (64, 70, 81 and 71%, respectively) and fresh weight increase (65, 41, 54 and 41%, respectively) compared to shoot culture growth with the Murashige-Skoog nitrogen mix of 40 mM nitrate and 20 mM ammonium. Glutamate and proline were at best poorly supportive, and glycine betaine and choline were non-supportive. Callus initiation from leaf discs was supported only by nitrate, ammonium, urea, glutamine and proline (50, 100, 100, 100 and 80%, respectively, at the best concentrations, of that on Murashige-Skoog medium). Subsequent shoot regeneration from the intact disc callus in those cultures only occurred on media with nitrate, urea, glutamine, or proline (12, 3, 28 and 3% as many shoots, respectively, as on Murashige-Skoog medium). Overall, the Murashige-Skoog nitrogen mix was superior or equal to any single nitrogen source. However, single nitrogen source media with nitrate, ammonium, urea, glutamine or proline should have significant utility for shoot or leaf disc cultures of mutants with impaired nitrogen assimilation, in comparative physiology studies, or in dual cultures with pathogens of limited ability to use any of these forms of nitrogen.

Synthesis of fluorochromes and pigments in Malassezia furfur by use of tryptophan as the single nitrogen source.

A new minimal medium consisting only of L-tryptophan (L-Trp) and a lipid source induced formation of brown pigmentation only in the species Malassezia furfur, which diffuses into the agar. Strains of the species M. sympodialis and M. pachydermatis failed to grow on this medium. On mDixon medium, however, after replacement of peptone by L-Trp, growth of all three Malassezia species was achieved. Under these conditions pigment production was observed with all M. furfur strains tested, although the results for M. pachydermatis strains were inconsistent. M. sympodialis strains showed no pigment production. On the minimal medium pigmentogenesis was induced in M. furfur by only 0.01 g% tryptophan; the pH optimum was pH 5. In all M. furfur strains, alternative amino nitrogen sources given concurrently with Trp suppressed pigmentogenesis. Furthermore, there were differences in the optimal temperature among the individual M. furfur strains. CBS 7019, CBS 6000 and CBS 6001 failed to produce pigment at 37 degrees C. The extract of the culture exhibited remarkable fluorescence, and several indole derivatives with a broad spectrum of colours were detected. This finding may have an impact on the clinical appearance of pityriasis versicolor, a very common skin disease caused by lipophilic yeasts of the genus Malassezia. We hypothesize that in pityriasis versicolor metabolic adaptation of Malassezia yeasts to altered nitrogen conditions on superficial skin might be of patho-physiological importance. Tryptophan as an inducer of pigmentogenesis probably accumulates during excessive sweating, a well-known manifestation of pityriasis versicolor.

Physiological and electron microscopical investigations on syntrophic dicyandiamide degradation by soil bacteria

Soil bacteria with high dicyandiamide (DCD) degrading capacity were isolated. In addition to pure cultures, two different consortia of soil bacteria were shown to degrade DCD under conditions of syntrophy. The single isolates of the consortia could not grow on minimal media or degrade DCD. These consortia turned out to be associations between Xanthomonas maltophilia and either Radiobacter sp. or Aureobacterium sp. Both types of consortia proliferate and degrade DCD rapidly. Supplementation experiments and microscopical investigations focusing on Xanthomonas maltophilia and Aureobacterium sp. showed that Aureobacterium sp. is probably auxotrophic for a yet unknown factor. Supplementation of minimal medium with minimum essential medium (MEM) vitamins enables this strain to grow with nitrate, but not with DCD or ammonia as single nitrogen sources. Xanthomonas maltophilia, however, is dependent on a substance which is not released into the medium by its companion. It cannot be provided either by the addition of vitamins or casamino acids or culture filtrates of proliferating consortia to the medium, but only by the presence of Aureobacterium sp. cells in the nutrient medium. Partially based on novel preparation techniques, microscopical investigations constantly showed a close contact between Xanthomonas maltophilia cells and its companion in enrichment medium, while Aureobacterium sp. is also found as single cells. The close contact appears to be of paramount importance to initiate growth and DCD degradation. The results demonstrate clearly the importance of investigations, including bacterial consortia in addition to pure culture studies when the degradability of agrochemicals has to be assessed.

Amino acid degradation by the mesophilic sulfate-reducing bacterium desulfobacterium vacuolatum

Desulfobacterium vacuolatum strain IbRM was able to grow using casamino acids as a source of carbon, energy and nitrogen. Growth was accompanied by utilization of several amino acids and sulfide production. Proline and glutamate were used preferentially and to the greatest extent. Glycine, serine and alanine were used more slowly and only after proline and glutamate were used. Isoleucine, valine, leucine and aspartate decrease was slowest and occurred in a linear fashion throughout the growth phase. Amino acids used from casamino acids, excluding aspartate, were also used as single carbon, energy and nitrogen sources. As a single amino acid, aspartate could only be used as a nitrogen source. Aspartate was not used as an electron acceptor. No growth occurred on any amino acid in the absence of sulfate. As single substrates, isoleucine, proline and glutamate were oxidized without formation of acetate and with molar yields of 13.1, 9.4 and 7.7 g mol-1, respectively.

Effect of Carbon, Nitrogen, and C:N Ratio on Growth, Sporulation, and Biocontrol Efficacy of Talaromyces flavus

ABSTRACT Engelkes, C. A., Nuclo, R. L., and Fravel, D. R. 1997. Effect of carbon, nitrogen, and C:N ratio on growth, sporulation, and biocontrol efficacy of Talaromyces flavus. Phytopathology 87:500-505.Five-day biomass production by the biocontrol fungus Talaromyces flavus was measured in a liquid basal medium, pH 5.5, containing each of 37 carbon (C) sources with a single nitrogen (N) source, and each of 42 N sources with a single C source. In general, production of biomass was greatest on complex sugars such as polysaccharides (32 g/liter of medium) and beta-glucosides (2.4 g/liter of medium), and was least on monosaccharides (1.3 g/liter of medium). Ascospore production at 6 weeks on solid basal medium with the same amount of these same 37 degrees C sources was greatest on oligosaccharides (2.9 x 10(8) spores per 5.5-cm-diameter petri dish), and least on polysaccharides and monosaccharides (1.6 and 1.4 x 10(8) spores per 5.5-cm-diameter petri dish, respectively). For C sources, there was no correlation between production of ascospores and hyphal dry weight. The various N sources yielded 0 to 10(9) ascospores per 5.5-cm-diameter petri dish and 10(-4) to 10(-5) g of hyphal dry weight per milliliter. In general, N sources that resulted in the greatest number of ascospores also yielded the greatest hyphal dry weights. For the two C and two N sources tested, the number of ascospores increased as the ratio of C to N increased from 5:1 to 30:1. This effect was most obvious as the C:N ratio increased from 5:1 to 15:1. At low C:N ratios (<15:1), treatments with hypoxanthine as a N source resulted in significantly greater production of biomass than treatments with ammonium tartrate; no difference was observed at C:N ratios >/=15:1. Incidence of Verticillium wilt was 50% lower for eggplants drenched with ascospores grown on potato dextrose agar (PDA) compared with eggplants either nondrenched or drenched with ascospores grown on media with hypoxanthine plus lactose or maltose. Thus, C and N sources that slightly increased ascospore production of T. flavus reduced efficacy of biocontrol of Verticillium wilt compared with ascospores produced on PDA.

Ammonia Inhibition of Plasmid pRmeGR4a Conjugal Transfer between Rhizobium meliloti Strains

We have examined nutritional factors influencing conjugal transfer of the two nonsymbiotic large plasmids, pRmeGR4a and pRmeGR4b, of Rhizobium meliloti GR4. To monitor transfer, each plasmid was tagged with a different antibiotic resistance marker. Transfer of plasmid pRmeGR4b was dependent upon the presence of plasmid pRmeGR4a on the same donor cell. Transconjugants for pRmeGR4b were obtained at frequencies 5-to 10-fold higher than transconjugants carrying both plasmids, indicating that mobilization of pRmeGR4b by pRmeGR4a probably occurred in trans. Conjugal transfer of the tagged plasmids between R. meliloti strains was tested on minimal medium supplemented with single amino acids, nitrate, or ammonium as the single nitrogen source. A higher number of transconjugants was obtained when glutamate was the only nitrogen source, whereas conjugation was virtually undetectable on ammonium. No relationship was found between donor or recipient growth rate and plasmid transfer rate on a given nitrogen source. Furthermore, in media containing both glutamate and ammonium as nitrogen sources, transfer was reduced almost 100-fold compared with that in media containing glutamate alone. Inhibition was readily detected at 2.5 mM or higher concentrations of either ammonium chloride or ammonium sulfate and appeared to be specific for exogenously supplied ammonium. Inhibition of conjugal transfer between R. meliloti strains by ammonium was only observed for rhizobial plasmids, not for a heterologous plasmid such as RP4. Apparently, ammonium did not affect the plasmid-encoded transfer machinery, as it had no influence on rhizobial plasmid transfer from R. meliloti to Agrobacterium tumefaciens. The effect of ammonium seemed to take place on R. meliloti recipient cells, thereby reducing the efficiency of plasmid conjugation, probably by affecting mating pair formation or stabilization.

Two distinct quinoprotein amine oxidases are induced by n-butylamine in the mycelia of Aspergillus niger AKU 3302. Purification, characterization, cDNA cloning and sequencing.

Two distinct quinoprotein amine oxidases were found in Aspergillus niger mycelia grown on n-butylamine medium and purified using chromatographic techniques. The respective enzymes were termed AO-I, which had already been isolated, and AO-II, a new enzyme found in this study. HPLC indicated that their molecular masses are 150 kDa and 80 kDa, respectively. On SDS/PAGE, the enzymes gave a similar but distinct mobility, which corresponds to 75 kDa for the subunit dimeric AO-I and 80 kDa for monomeric AO-II. The absorption spectra of both enzymes were different from each other; the absorption maxima in the visible region were at 490 nm for AO-I and 420 nm for AO-II. The enzymes showed positive quinone staining, comparable substrate specificity, and sensitivity to inhibitors typical for copper/topa quinone-containing amine oxidases, but they had different copper contents and also differed in their N-terminal sequences. Their peptide maps showed almost identical patterns, with the exception of two additional bands for AO-II. Among the peptides obtained from digestion of AO-II, peptides with sequences corresponding to the N-terminal part of AO-I were detected. Polyclonal antibodies raised against AO-I and AO-II recognized both enzymes, but with different specificities. Using precipitation with AO-I, the antibody prepared against AO-II was purified and was shown to be specific only for AO-II. The cDNA of AO-I was cloned and sequenced. A highly conserved tetrapeptide sequence, Asn-Tyr-Glu-Tyr, was identified in which the first tyrosine residue (Tyr404) that could be converted to topa quinone was present in the 670-residue deduced amino acid sequence. Northern blot analysis indicated that AO-I was highly expressed in A. niger grown on n-butylamine as a single nitrogen source. Genomic Southern blot analysis confirmed that both enzymes are likely to be encoded by the same gene.

Variation in Nitrogen Utilization in Acremonium coenophialum Isolates

Acremonium coenophialum, a fungal endosymbiont in tall fescue, is responsible for the production of alkaloid toxins. Animals grazing endophyte-infected tall fescue often show toxicosis. In marginal environments, the endophyte is important for long-term survival of tall fescue. Few differences in endophyte isolates from individual tall fescue plants have been reported. To aid development of a toxicosis-free tall fescue, it is important to identify differences in endophyte isolates. This report describes variation in nitrogen utilization in a defined culture medium by endophyte isolates from Kentucky-31 tall fescue. Overall, the best nitrogen sources for dry weight production of mycelium were proline and potassium nitrate. Thirty-four isolates grown on agar-solidified defined media with single nitrogen sources showed variation in nitrogen utilization. Fifty percent of the isolates were unable to utilize two or more amino acids. Manipulation of endophyte variation could lead to development of a toxicosis-free tall fescue cultivar.

Increase in Nitrate Reductase Activity with Ammonium Chloride inBacillus licheniformisby Shaking Culture

In shaking culture, nitrate reductase activity in the cell-free extracts of Bacillus licheniformis increased with the addition of NH4Cl to the medium containing NaNO3 as a single nitrogen source, where amounts of nitrogen sources were sufficient for cell growth. This increase of nitrate reductase activity therefore suggests that the activity is not for nitrate assimilation but for other physiological functions containing a dissimilatory nitrate reduction.

Mycobacterium cookii sp. nov.

Strains of a new type of slowly growing scotochromogenic mycobacterium were isolated repeatedly from sphagnum vegetation and surface water of moors in New Zealand. These strains grew at 31 and 22 degrees C but not at 37 degrees C and possessed catalase, acid phosphatase, and arylsulfatase activities. They did not split amides, and most of them were susceptible to antituberculotic drugs. Furthermore, they did not tolerate 0.1% NaOH2 and 0.2% picric acid and did not grow on compounds used as single carbon sources and single nitrogen and carbon sources. The internal similarity of the strains as determined by numerical taxonomy methods was 96.6% +/- 3.09%. The whole-mycolate pattern is unique in that it has not been found previously in 23 species of slowly growing mycobacteria. Evaluation of long-reverse-transcriptase-generated stretches of the primary structure of the 16S rRNA confirmed that these organisms belong to the genus Mycobacterium. The phylogenetic position of these bacteria is unique; they are situated between slowly growing pathogenic and rapidly growing saprophytic species. The strains are not pathogenic for mice, guinea pigs, and rabbits, but they provoke a nonspecific hypersensitivity reaction to bovine tuberculin. Hence, they are considered members of a new species of nonpathogenic, slowly growing mycobacteria, for which the name Mycobacterium cookii is proposed. Strain NZ2 is the type strain; a culture of this strain has been deposited in the American Type Culture Collection as strain ATCC 49103.

Cultivation of artemisia annua L plantlets in a bioreactor containing a single carbon and a single nitrogen source

Micropropagation of a number of plant species by the shoot culture technique is well established. The process is labor intensive and expensive. Micropropagation in submerged culture systems are potentially cost-effective as less materials and labor may be required and shoots and roots may be grown in the same reactor vessel

Chitosanase from Streptomyces griseus

Publisher Summary This chapter describes the assay method and procedure for the purification of chitosanase from Streptomyces griseus. The assay is based on the estimation of amino sugars produced in the hydrolysis of glycol chitosan, a water-soluble derivative of chitosan, by the method of Rondle and Morgan, s using glucosamine as a reference compound. The purified chitosanases are classified into two groups: the enzymes hydrolyzing only chitosan and the enzymes hydrolyzing both chitosan and carboxymethylcellulose. A strain of Streptomyces griseus has produced chitosanase in culture broth with chitosan as the single carbon and nitrogen source. The purified enzyme is able to hydrolyze chitosan and carboxymethylcellulose, and produces glucosamine oligomers in the hydrolysis of chitosan.