L. F. Linhares

Chitinolytic activity of actinomycetes from a cerrado soil and their potential in biocontrol

R.C. GOMES, L.T.A.S. SEME(r)DO, R.M.A. SOARES, C.S. ALVIANO, L.F. LINHARES and R.R.R. COELHO.2000.The crude enzyme extracts from five actinomycetes selected from a cerrado soil presented very good endochitinolytic activity when compared to a commercial chitinase. Exochitinase and chitobiase activities were also detected. They were identified as Streptomyces, but could not be characterized to species level, probably corresponding to new ones. The crude extracts, obtained from growth on fungal mycelium plus chitin of three of the strains, have shown a very pronounced activity against phytopathogenic fungi. In tests using growing cells, all five strains were active. These data suggest that these strains are potential biocontrol agents.

Characterization of fungal melanins and soil humic acids by chemical analysis and infrared spectroscopy.

SummaryHumic acids from two Brazilian topsoils under savanna grassland and five soil fungal melanins were characterized by elemental, functional group and infrared analysis. C, N, total acidity, COOH, and phenolic OH contents were within the ranges reported for several other fungal melanins and soil humic acids. Compared with the soil humic acids, the infrared spectra of the fungal melanins showed greater detail, indicative of higher aliphaticity. They were similar to the type III infrared spectra of humic acids, which are characteristically high in proteinaceous material and polysaccharides. The infrared spectra of the humic acids from the two Brazilian soils studied were classified as type I, which includes most soil humic acids. Notwithstanding the greater detail, in some areas the fungal melanin spectra were similar to those reported for other fungal melanins and humic acids of different origins. The probable contribution of the melanic fungi to the formation of soil humic polymers is discussed.

Isolation and characterization of actinomycetes from Brazilian tropical soils

Actinomycetes have been isolated from three Brazilian tropical soils. The dispersion and differential centrifugation procedure revealed count values 1.5 to 5.0 times greater than those obtained by the conventional dilution plate technique for all soils and media tested. Eighteen strains, promising for biotechnological applications, were submitted to chemotaxonomic procedures and numerical taxonomy for identification. Two were identified as Amycolatopsis orientalis, one as Streptomyces misakiensis, and two tentatively included or associated to S. chromofuscus and S. griseoruber. The others, all belonging to the Streptomyces genus, could not be fitted into any known species, and were arranged by the UPGMA analysis for classification, as an isolated group. This suggests that the actinomycetes in tropical soils may represent a vast unexplored resource for biotechnology.

Amino acid distribution in some fungal melanins and of soil humic acids from Brazil

SummaryHumic acids from four Brazilian topsoils of different origins and four soil fungal melanins, synthesized under two cultural conditions, were subjected to 6N HCl hydrolysis and their amino acid distribution patterns qualitatively and quantitatively determined. Both soil and fungal polymers showed similar patterns with aspartic acid, glutamic acid, glycine and alanine as the dominant amino acids. Some variations noted were more quantitative than qualitative, the similarities were more pronounced than differences, indicating that the fungal melanins may play a significant role in the formation of soil humic acid polymers. The humic acids of Brazilian soils had amino acid distribution patterns similar to those reported for humic acids of other tropical and temperate soils.

Elemental, functional group and infrared spectroscopic analysis of actinomycete melanins from brazilian soils

Eleven actinomycete melanins were characterized by elemental and functional group and infrared analysis. A soil humic acid from a Brazilian topsoil, a darkred latosol under savanna grassland, analysed previously, was used for comparative purposes. C, N, total acidity, COOH, and phenolic OH contents were within the ranges reported for soil humic acids and fungal melanins. Compared to the soil humic acid, the actinomycete melanins showed greater detail, indicative of higher aliphaticity. Most of these were, in varying degrees, similar to the type III IR spectra of humic acids, which are characteristically high in proteinaceous material, and with variable amounts of polysaccharides. The exceptions were two melanin spectra that showed more resemblance to the humic acid from the dark-red latosol, which belongs to the type I spectra of soil humic acids, a category that includes most soil humic acids. The probable participation of melanic actinomycetes in the formation of humic polymers in discussed.

Sugars in hydrolysates of fungal melanins and soil humic acids

Humic acids from four Brazilian topsoils of different origin and four fungal melanins, synthesized under two cultural conditions were subjected to a two step hydrolysis procedure and the released monosaccharides qualitatively and quantitatively determined by gas-liquid chromatography. The neutral sugars, glucose, galactose, mannose, arabinose, xylose, fucose, rhamnose and the alcohol sugar inositol, were detected in most of the soil humic acid samples. The fungal melanins showed the presence of glucose, galactose, mannose and arabinose. Ribose was present in two out of the eight samples tested. Some quantitative differences in the two types of humic polymers were noted and expected considering their origins. However, similarities were more apparent than differences and give further indication that melanic fungi may play a significant role in the formation of soil humic acids.

Chitinolytic actinomycetes from a Brazilian tropical soil active against phytopathogenic fungi

Five Streptomyces spp. isolated from a Brazilian forest soil showed endochitinase activity in the alkaline range with optima between 40 and 50 °C. Three were highly active against three phytopathogenic fungi by in vitro experiments. Preliminary experiments suggested that the isolates may belong to a new taxon.

Melanogenic actinomycetes (Streptomyces spp.) from Brazilian soils

SummaryMelanogenic actinomycetes were isolated from cerrado soils. Starch agar with a neutral pH was the best medium for selecting pigment-producing colonies. A pigmentation screening test selected 52% of these as possible melanin producers. Tests on liquid (organic and inorganic) and solid (peptone and tyrosine) media, and enzymatic tests, confirmed about 90% as melanin producers, 68% as dihydroxyphenylalanine (DOPA)-melanin and 32% as possible other kinds of melanin producers. Melanin production occurred mostly with an organic N, or an inorganic N with an additional organic N source. An exception was observed with three strains, which were able to produce melanins with an entirely inorganic N source in the medium. Instability of melanin production was a common feature in many strains. Further characterization of melanins produced by actinomycetes compared with soil humic acids may clarify the possible role of melanogenic actinomycetes in soil organic matter formation.

Efficiency of the dispersion and differential centrifugation technique in the isolation of chitinolytic actinomycetes from soil

Actinomycetes were isolated from an acidic Brazilian soil under cerrado (savanna) vegetation, previously amended with chitin. The efficiency of two isolation techniques, the conventional dilution plate and the dispersion and differential centrifugation (DDC) procedures, were compared and a mathematical computer analysis of the results was made. The DDC technique gave counts about nine times greater than the conventional one. Analysis of the derivative curves suggested the presence of two distinct groups of actinomycetes on the isolation plates, one able to induce chitinolytic enzymes during amendment and the other whose initial activity was delayed. Major differences in results with different isolation procedures suggest a more subtle ecological role for these microbes in terms of their location in the soil environment.

Neutral sugars in melanins synthesized by actinomycetes from Brazilian soils

Abstract Nine actinomycete melanins synthesized under various culture conditions, eight of them by actinomycete samples isolated from Brazilian topsoils under savanna (cerrado) vegetation and one from an ATCC sample, were subjected to a two-step hydrolysis procedure and the sugars released qualitatively and quantitatively determined by capillary gas-liquid chromatography (GLC). Humic acids (HAs) extracted from these soils, analysed previously, were used for comparison. The neutral sugars glucose, galactose, mannose, xylose, arabinose, ribose, rhamnose and fucose and the alcohol sugar inositol were present in varying amounts in most of the melanins analysed. The same sugars were present in the HAs used for comparison, except for ribose. Some qualitative and quantitative differences observed in the two types of macromolecules would be expected, considering their origins. The results indicate that the actinomycete melanins have a qualitative sugar distribution pattern similar to that of the HAs from Brazilian tropical soils and of HAs reported for soils from other climatic regions. The possible participation of actinomycete melanins in the formation of soil humic substances is discussed.