Allostreptomyces indica sp. nov., isolated from India.

A novel actinobacterium, designated strain YIM 75704T, was isolated from a limestone quarry located at Gulbarga, Karnataka, India. The novel strain has showed typical morphological and chemotaxonomic characteristics of the family Streptomycetaceae. Comparison of 16S rRNA gene sequences indicated that this strain represents a novel member of the family Streptomycetaceae and exhibited 99.0% 16S rRNA gene sequence similarities with the type species of the recently described novel genus Allostreptomyces, that is, Allostreptomyces psammosilenae, whereas other species of Streptomyces were below 95% sequence similarity. The cell hydrolysates contained the LL-isomer of diaminopimelic acid and the predominant quinones were MK-9 (H6, H8 and H4). The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylinositolmannosides and three unknown phospholipids. The DNA G+C content was 75.0 mol%. A polyphasic study of the strain with morphological, phenotypic, phylogenetic and with DNA-DNA hybridization evidence with related members showed that this strain represents novel species of Allostreptomyces for which the name Allostreptomyces indica sp. nov., is proposed. The type strain is YIM 75704T (= DSM 41985T=CCTCC AA 209051T= NCIM 5485T).

Production, characterization and application of keratinase from Streptomyces gulbargensis.

A Streptomyces gulbargensis newly isolated, thermotolerant feather-degrading bacterial strain was investigated for its ability to produce keratinase enzyme. Maximum keratinolytic activity was observed at 45 degrees C and pH 9.0 at 120 h of incubation. Activity was completely stable (100%) between 30 and 45 degrees C and pH 7.0-9.0, respectively. Addition of starch to the growth medium affects the activity by means of increase in keratinase secretion. After seven days of cultivation, 10-fold increase (14.3 U ml(-1)) in keratinase activity was observed in the presence of 3g starch (per liter) of the medium. The enzyme was monomeric and had a molecular mass of 46 kDa. The enzyme activity was significantly inhibited by CaCl(2) and partly inhibited by EDTA, whereas, Na(2)SO(3) enhance the enzyme activity by 2.9 times more. In addition, native chicken feather was completely degraded at 96 h of incubation. The results obtained showed that newly isolated strain S. gulbargensis could be a useful in biotechnology in terms of valorization of keratin-containing wastes or in the leather industry.

Production and partial purification of alpha-amylase from a novel isolate Streptomyces gulbargensis.

Extracellular amylase production by a newly isolated alkali-thermotolerant strain Streptomyces gulbargensis DAS 131 was optimized and characterized. The highest amylase production was achieved by growing S. gulbargensis DAS 131 in media with 1% starch. Strain exhibited maximal activity at pH 9.0 and 45 degrees C and relatively stable in alkaline conditions (pH 11). Starch and peptone were found to be the good source of carbon and nitrogen with a yield of 2,216.6 and 2,156.1 U, respectively. Maltose and maltotriose were the main end products of starch hydrolysis, indicating alpha-amylase activity. SDS-PAGE analysis revealed a monomeric form with a molecular weight of 55 kDa.

Streptomyces deccanensis sp. nov., an alkaliphilic species isolated from soil.

A novel actinomycete strain, DAS-139T, was isolated from a soil sample collected from Gulbarga, Karnataka Province, India. The isolate was characterized by white to grey aerial mycelium. Long spore chains were found on the aerial mycelium and the aerial mycelium was composed of non-motile spores with hairy surfaces. The cell wall of strain DAS-139T contained ll-diaminopimelic acid isomer as the diagnostic diaminoacid indicating that the cell wall was of chemotype-I. The predominant menaquinones were MK-9(H6) (76 %), MK-9(H4) (14 %) and MK-9(H8) (10 %). Phosphatidylethanolamine was the diagnostic phospholipid. On the basis of 16S rRNA gene sequence, phenotypic and phylogenetic analyses, the novel strain was identified as a member of the genus Streptomyces. The novel strain grew optimally at 28 degrees C and pH 9.0. The G+C content of the genomic DNA was 71.8 mol%. 16S rRNA gene sequence analysis showed that the novel isolate had 99.4 % sequence similarity with Streptomyces scabiei ATCC 49173T and 99.2 % similarity with Streptomyces diastachromogenes ATCC 12309T. Furthermore, DNA-DNA hybridization with these two Streptomyces species showed 36.0 and 43.0 % relatedness, respectively. Based on these observations, strain DAS-139T is proposed to represent a novel species of the genus Streptomyces, for which the name Streptomyces deccanensis sp. nov. is proposed with the type strain DAS-139T (=KCTC 19241T=CCTCC AA 207004T).

Saccharomonospora saliphila sp. nov., a halophilic actinomycete from an Indian soil.

A Gram-positive, moderately halophilic actinomycete strain, designated YIM 90502(T), was isolated from a sample of muddy soil collected from Gulbarga, Karnataka Province, India, and subjected to a polyphasic taxonomical analysis. The isolate grew optimally at 28 degrees C and in the presence of 10 % (w/v) NaCl. The isolate was characterized chemotaxonomically as having meso-diaminopimelic acid in the cell-wall peptidoglycan and galactose and arabinose as whole-cell sugars. The predominant menaquinone was MK-9(H(4)), while MK-8(H(4)) was found in smaller amounts. The phospholipids were phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine. The major fatty acids were iso-C(16 : 0) (49.2 %) and C(17 : 1)omega6c (9.1 %). The DNA G+C content of the isolate was 71.8 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that the isolate fell within the evolutionary radiation encompassed by the genus Saccharomonospora. 16S rRNA gene sequence similarity between strain YIM 90502(T) and the type strains of Saccharomonospora species ranged from 92.42 % (with Saccharomonospora xinjiangensis CCTCC AA 97021(T)) to 97.45 % (with Saccharomonospora azurea KCTC 9693(T)). Levels of DNA-DNA relatedness between strain YIM 90502(T) and S. azurea KCTC 9693(T), Saccharomonospora halophila DSM 44411(T) and Saccharomonospora paurometabolica DSM 44619(T) were 46.0, 41.0 and 42.5 %, respectively. On the basis of phenotypic, phylogenetic and genotypic data, strain YIM 90502(T) was classified in the genus Saccharomonospora as a member of a novel species, for which the name Saccharomonospora saliphila sp. nov. is proposed, with YIM 90502(T) (=KCTC 19234(T) =DSM 45087(T)) as the type strain.

Streptomyces tritolerans sp. nov., a novel actinomycete isolated from soil in Karnataka, India.

A Gram-positive, nonmotile, moderately halophilic, alkali and thermotolerant strain designated DAS 165(T), was isolated from a dry land soil sample from the Gulbarga region, Karnataka province, India. The isolate produced yellow substrate mycelia and gray aerial mycelia on most tested media. Strain DAS 165(T) showed growth in the presence of 5 to 7% NaCl and at 45 degrees C. The DNA G + C content was 69.7%. 16S rRNA gene sequence analysis together with these characteristics consistently assigned strain DAS 165(T) to the genus Streptomyces. The 16S rRNA gene sequence analysis revealed that strain DAS 165(T) was most closely related to S. tendae ATCC 19812(T) (D 63873) with a sequence similarity of 99.6% (three nucleotide differences out of 1,517). Strain DAS 165(T) formed a distinct clade based on analysis of the almost complete sequence and 120-nucleotide variable gamma region of the 16S rRNA gene. Despite the high sequence similarity, strain DAS 165(T) was phenotypically different from S. tendae ATCC 19812(T). DNA-DNA hybridization between these strains was 47% showing that strain DAS 165(T) is a distinct genomic species. Phenetic and genetic results support the classification of strain DAS 165(T) as a new species, for which the name S. tritolerans is proposed, with strain DAS 165(T) as the type strain (=DSM 41899(T )= CCTCCAA 206013(T)).

Streptomyces gulbargensis sp. nov., isolated from soil in Karnataka, India.

During the course of screening for industrially important microorganisms, an alkali-tolerant and thermotolerant actinomycete, strain DAS 131T, was isolated from a soil sample collected from the Gulbarga region, Karnataka province, India. The strain was characterized by a polyphasic approach that showed that it belonged to the genus Streptomyces. Growth was observed over a wide pH range (pH 6-12) and at 45 degrees C. The 16S rRNA gene sequence of strain DAS 131T was deposited in the GenBank database under the accession number DQ317411. 16S rRNA gene sequence analysis revealed that strain DAS 131T was most closely related to Streptomyces venezuelae ISP 5230T (AY999739) with a sequence similarity of 99.5% (8 nucleotide differences out of 1,477). Despite this very high sequence similarity, strain DAS 131T was phenetically distinct from S. venezuelae. The DNA relatedness between these strains was 54%, indicating that strain DAS 131T is a distinct genomic species. On the basis of phenetic and genetic analyses, strain DAS 131T is classified as a new species in the genus Streptomyces, for which we propose the name Streptomyces gulbargensis sp. nov.