Identification by molecular techniques of halophilic bacteria producing important enzymes from pristine area in Campeche, Mexico / Identificação por técnicas moleculares de bactérias halofílicas que produzem enzimas importantes em área intocada de Campeche, México

Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Isla Arena está localizada na coordenada 20°70’N - 90°45’W, de Campeche, México. Nesses estuários, o oceano se mistura com a água doce e os ecossistemas se concentram onde proliferam petenos e flamingos rosa. Crustáceos e moluscos abundam no mar. Apesar de sua enorme riqueza marinha, não há estudos realizados sobre a presença de microrganismos halofílicos nessas águas. Neste trabalho, a diversidade e estrutura da comunidade microbiana foram investigadas através de uma abordagem metagenômica e corroboradas para o sequenciamento de genes 16S rRNA. Verificou-se que o filo Fimicutes predomina com mais de 50%, quase na mesma proporção da classe Bacilli e com quase 41% de abundância relativa da ordem Bacillales. Os resultados do sequenciamento mostraram que uma das amostras apresentou alto percentual de similaridade (99,75%) pelo programa Nucleotide BLAST com um microrganismo peculiar: Bacillus subtilis. Nossos resultados demonstram que B. subtilis pode ser uma fonte eficiente de proteases, lipases e celulases, provenientes de comunidades microbianas halofílicas localizadas em áreas pouco exploradas.

Identification by molecular techniques of halophilic bacteria producing important enzymes from pristine area in Campeche, Mexico

Abstract Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Resumo Isla Arena está localizada na coordenada 20°70N - 90°45W, de Campeche, México. Nesses estuários, o oceano se mistura com a água doce e os ecossistemas se concentram onde proliferam petenos e flamingos rosa. Crustáceos e moluscos abundam no mar. Apesar de sua enorme riqueza marinha, não há estudos realizados sobre a presença de microrganismos halofílicos nessas águas. Neste trabalho, a diversidade e estrutura da comunidade microbiana foram investigadas através de uma abordagem metagenômica e corroboradas para o sequenciamento de genes 16S rRNA. Verificou-se que o filo Fimicutes predomina com mais de 50%, quase na mesma proporção da classe Bacilli e com quase 41% de abundância relativa da ordem Bacillales. Os resultados do sequenciamento mostraram que uma das amostras apresentou alto percentual de similaridade (99,75%) pelo programa Nucleotide BLAST com um microrganismo peculiar: Bacillus subtilis. Nossos resultados demonstram que B. subtilis pode ser uma fonte eficiente de proteases, lipases e celulases, provenientes de comunidades microbianas halofílicas localizadas em áreas pouco exploradas.

Identification by molecular techniques of halophilic bacteria producing important enzymes from pristine area in Campeche, Mexico / Identificação por técnicas moleculares de bactérias halofílicas que produzem enzimas importantes em área intocada de Campeche, México

Abstract Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Resumo Isla Arena está localizada na coordenada 20°70'N - 90°45'W, de Campeche, México. Nesses estuários, o oceano se mistura com a água doce e os ecossistemas se concentram onde proliferam petenos e flamingos rosa. Crustáceos e moluscos abundam no mar. Apesar de sua enorme riqueza marinha, não há estudos realizados sobre a presença de microrganismos halofílicos nessas águas. Neste trabalho, a diversidade e estrutura da comunidade microbiana foram investigadas através de uma abordagem metagenômica e corroboradas para o sequenciamento de genes 16S rRNA. Verificou-se que o filo Fimicutes predomina com mais de 50%, quase na mesma proporção da classe Bacilli e com quase 41% de abundância relativa da ordem Bacillales. Os resultados do sequenciamento mostraram que uma das amostras apresentou alto percentual de similaridade (99,75%) pelo programa Nucleotide BLAST com um microrganismo peculiar: Bacillus subtilis. Nossos resultados demonstram que B. subtilis pode ser uma fonte eficiente de proteases, lipases e celulases, provenientes de comunidades microbianas halofílicas localizadas em áreas pouco exploradas.

Bacterias halófilas con potencial para la recuperación de suelos salinizados en Sáchica-Boyacá, Colombia

Es posible usar bacterias halófilas aisladas de ambientes salinos como una posible alternativa de rehabilitación de suelos salinizados para la producción agrícola y agropecuaria. En este estudio se evaluaron seis bacterias halófilas para determinar su potencial en la disminución de la conductividad eléctrica (Ce) de suelo moderadamente salino del municipio de Sáchica- Boyacá (Colombia). Las bacterias se aislaron en agar nutritivo con diferentes concentraciones de NaCl, luego fueron inoculadas en el suelo (3 kg puesto en materas) para realizar un bioensayo en invernadero durante 2 meses. Se realizaron dos fases, la primera hasta conseguir un valor de 2 ds/m (Ce) con el fin de sembrar plantas de tomate en la segunda fase. Se realizaron mediciones semanales de la (Ce) durante todo el bioensayo y solo en la fase 2 se determinaron semanalmente variables cualitativas de crecimiento de tomate (altura, número de hojas, área foliar, marchitez y clorosis). Los resultados mostraron un efecto directo de las bacterias halófilas en la disminución de la (Ce) del suelo, pasando de 5.2 a 0.8 ds/m (T1), 0.7 ds/m (T2), 0.8 ds/m (T3), 0.6 ds/m (T4), 0.7 ds/m (T5), 0.7 ds/m (T6), 3.0 ds/m (testigo), valores que corresponden a salinidad muy ligera (< 2 ds/m). En relación con las variables agronómicas, se evidenciaron mejoras en las características evaluadas en comparación con el testigo absoluto, sobre todo en altura y número de hojas de las plantas. Para el testigo absoluto se perdieron 80 % de las unidades experimentales, mientras que en el suelo con los diferentes tratamientos solo en algunos casos se perdió el 20 %. De acuerdo con esto, se evidencia que las bacterias halófilas utilizadas en esta investigación tienen potencial para la rehabilitación de suelos salinizados.

It is possible to use halophilic bacteria isolated from saline environments as a possible alternative for the rehabilitation of salinated soils for agricultural and agricultural production. Six halophilic bacteria were evaluated to determine their potential in the reduction of the electrical conductivity (Ce) of moderately saline soil in the municipality of Sáchica-Boyacá (Colombia). The bacteria were isolated on nutritive agar with different concentrations of NaCl, then inoculated in the soil (3 kg placed in pots) to perform a bioassay in the greenhouse for 2 months. Two phases were carried out, the first until obtaining a value of 2 ds / m (Ce) in order to plant tomato plants in the second phase. Weekly measurements of the ( Ce ) were made throughout the bioassay and only in phase 2 were determined weekly qualitative variables of tomato growth (height, number of leaves, foliar area, wilting and chlorosis). The results showed a direct effect of the halophilic bacteria in the decrease of the (Ce) of the soil, going from 5.2 to 0.8 ds / m (T1), 0.7 ds / m (T2), 0.8 ds / m (T3), 0.6 ds / m (T4), 0.7 ds / m (T5), 0.7 ds / m (T6), 3.0 ds / m (Control), values that correspond to very light salinity (< 2 ds / m). Regarding the agronomic variables, improvements were observed in the characteristics evaluated in comparison with the absolute control, especially in height and number of leaves of the plants. For the absolute control 80 % of the experimental units were lost, while in the soil with the different treatments only in some cases 20 % was lost. Accordingly, it is evident that the halophilic bacteria used in this research have potential for the rehabilitation of salinated soils.

Bioprospection of proteases from Halobacillus andaensis for bioactive peptide production from fish muscle protein

BACKGROUND: Biologically active peptides produced from fish wastes are gaining attention because their health benefits. Proteases produced by halophilic microorganisms are considered as a source of active enzymes in high salt systems like fish residues. Hence, the aim of this study was the bioprospection of halophilic microorganisms for the production of proteases to prove their application for peptide production. RESULTS: Halophilic microorganisms were isolated from saline soils of Mexico and Bolivia. An enzymatic screening was carried out for the detection of lipases, esterases, pHB depolymerases, chitinases, and proteases. Most of the strains were able to produce lipases, esterases, and proteases, and larger hydrolysis halos were detected for protease activity. Halobacillus andaensis was selected to be studied for proteolytic activity production; the microorganism was able to grow on gelatin, yeast extract, skim milk, casein, peptone, fish muscle (Cyprinus carpio), and soy flour as protein sources, and among these sources, fish muscle protein was the best inducer of proteolytic activity, achieving a protease production of 571 U/mL. The extracellular protease was active at 50°C, pH 8, and 1.4 M NaCl and was inhibited by phenylmethylsulfonyl fluoride. The proteolytic activity of H. andaensis was used to hydrolyze fish muscle protein for peptide production. The peptides obtained showed a MW of 5.3 kDa and a radical scavenging ability of 10 to 30% on 2,2-diphenyl-1-picrylhydrazyl and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) and a ferric reducing ability of plasma. Conclusion: The use of noncommercial extracellular protease produced by H. andaensis for biologically active peptide production using fish muscle as the protein source presents a great opportunity for high-value peptide production.

Diversity of culturable moderately halophilic bacteria producing extracellular hydrolytic enzymes from marine sediments / Diversidade de bactérias cultiváveis moderadamente halofílicas que produzem enzimas hidrolíticas extracelulares de sedimentos marinhos

A total of 114 moderately halophilic bacteria were isolated from marine sediment environments. The isolates are belonged to 23 species based on the 16S rRNA sequence analysis. 63, 52, 47, 57, 74, 15 and 4 isolates are able to produce protease, amylase, lipase, pectinase, pulluanase, xylanase, cellulase, respectively. Combined hydrolytic enzyme activity analysis show that 15 strains present 1 hydrolytic activity, 32 strains present 2 hydrolytic activities, 21 strains present 3 hydrolytic activities, 26 strains present 4 hydrolytic activities, 11 strains present 5 hydrolytic activities and 2 strains present 6 hydrolytic activities. Hydrolase activities are widely distributed in a variety of species. The highest rates for production of protease, amylase, lipase, pectinase, pullanase, xylanase and cellulase were observed in species of B. baekryungensis, Hallobacillus sp., B. pumilus, B. megaterium or P. chungwhensis, B. amyloliquefaciens, B. pumilus, B. baekryungensis, respectively. However, the higher activities of protease, pectinase and pulluanase are frequently produced by the species of Halomonas sp. B. amyloliquefaciens or P. chungwhensis, and Vibrio sp. respectively. This investigation show that the diversity of halophilic bacteria from marine sediments could serve as a potential source of hydrolytic enzymes for industrial applications. (AU)

Um total de 114 bactérias moderadamente halofílicas foram isoladas de ambientes de sedimentos marinhos. Os isolados pertencem a 23 espécies com base na análise da sequência 16S rRNA. 63, 52, 47, 57, 74, 15 e 4 isolados são capazes de produzir protease, amilase, lipase, pectinase, pululanase, xilanase, celulase, respectivamente. A análise da atividade enzimática hidrolítica combinada mostra que 15 cepas apresentam 1 atividade hidrolítica, 32 cepas apresentam 2 atividades hidrolíticas, 21 cepas apresentam 3 atividades hidrolíticas, 26 cepas apresentam 4 atividades hidrolíticas, 11 cepas apresentam 5 atividades hidrolíticas e 2 cepas apresentam 6 atividades hidrolíticas. Atividades de hidrolase são amplamente distribuídas em uma variedade de espécies. As maiores taxas de produção de protease, amilase, lipase, pectinase, pululanase, xilanase e celulase foram observadas em espécies de B. baekryungensis, Hallobacillus sp., B. pumilus, B. megaterium ou P. chungwhensis, B. amyloliquefaciens, B. pumilus, B. baekryungensis, respectivamente. No entanto, as atividades mais elevadas de protease, pectinase e pululanase são freqüentemente produzidas pelas espécies de Halomonas sp. B. amyloliquefaciens ou P. chungwhensis e Vibrio sp. respectivamente. Esta investigação mostra que a diversidade de bactérias halofílicas de sedimentos marinhos pode servir como uma fonte potencial de enzimas hidrolíticas para aplicações industriais. (AU)

Proteolytic activity of recombinant DegP from Chromohalobacter salexigens BKL5

Background: DegP is a serine protease that specifically cleaves and refolds unfolding proteins in the periplasmic space of the cells. To date, there is no information regarding DegP from halophilic bacteria. Chromohalobacter salexigens BKL5 is a moderately halophilic bacterium that has the ability to grow in a media containing more than 15% salt. Therefore, the objectives of this work were to clone and overexpress DegP-encoding gene from C. salexigens BKL5 and characterize its biochemical properties. Results: DegP-encoding gene was overexpressed in Escherichia coli BL21(DE3) CodonPlus in an active form. SDS-PAGE analysis showed that the molecular weight of the recombinant DegP was 45 kDa. Size-exclusion chromatography analysis suggested that recombinant DegP was present in two multimeric states, hexameric and dodecameric, with molecular weights of 297.9 and 579.12 kDa, respectively. Both conformations were enzymatically active when casein was used as substrate for enzymatic assay. Circular dichroism analysis showed that recombinant DegP was composed of 0.21­0.29 helical content, which was comparable to the helical content in the crystal structure of E. coli DegP. The basic/acidic residue ratio of recombinant DegP was 0.56, which was slightly higher than that of DegP from extreme halophiles (average, 0.45) but significantly lower than that of DegP from nonhalophiles (average, 0.94). Conclusions: Recombinant DegP from C. salexigens BKL5 showed proteolytic activity when ß-casein was used as a substrate. In silico analysis indicated that recombinant DegP had characteristics similar to those of halophilic proteins depending on its amino acid composition.

Culturable diversity of halophilic bacteria in foreshore soils

Halophilic bacteria are commonly found in natural environments containing significant concentration of NaCl such as inland salt lakes and evaporated sea-shore pools, as well as environments such as curing brines, salted food products and saline soils. Dependence on salt is an important phenotypic characteristic of halophilic bacteria, which can be used in the polyphasic characterization of newly discovered microorganisms. In this study the diversity of halophilic bacteria in foreshore soils of Daecheon, Chungnam, and Saemangeum, Jeonbuk, was investigated. Two types of media, namely NA and R2A supplemented with 3%, 5%, 9%, 15%, 20% and 30% NaCl were used. More than 200 halophilic bacteria were isolated and BOX-PCR fingerprinting analysis was done for the typing of the isolates. The BLAST identification results showed that isolated strains were composed of 4 phyla, Firmicutes (60%), Proteobacteria (31%), Bacteriodetes (5%) and Actinobacteria (4%). Isolates were affiliated with 16 genera and 36 species. Bacillus was the dominant genus in the phylum Firmicutes, comprising 24% of the total isolates. Halomonas (12%) and Shewanella (12%) were also found as the main genera. These findings show that the foreshore soil of Daecheon Beach and Saemangeum Sea of Korea represents an untapped source of bacterial biodiversity.

Production of Extracellular Enzymes by Halophilic Bacteria Isolated from Solar Salterns.

During the course of survey of halophilic microorganisms, a total of sixteen bacterial isolates were obtained from coastal solar salterns of Orissa and West Bengal, India. Morphological, physiological and biochemical characteristics of these isolates indicate that majority of them belong to the genus Halomonas, however, members belonging to Cobetia and Halococcus were not uncommon. These isolates were screened for the production of extracellular enzymes such as amylase, glutaminase, asparaginase, xylanase, cellulase, gelatinase, inulinase, caseinase, pectinase, urease and lipase. Among these hydrolytic enzymes, glutamine and asparagine hydrolytic activities were predominant, although lipid and casein degrading activities were not inferior. However, amylase and gelatinase production were rare. None of these halophiles was able to degrade cellulose, inulin, pectin and xylan and only one isolate was capable of hydrolyzing urea.

Isolation and Characterization of Secondary Metabolites from Halophilic Bacillus Species from Marin drive in Mumbai.

The present study was focussed on production of novel antibiotics from Halophilic bacterial species in Marin drive Mumbai. Bacillus pumilus were isolated from soil and screened for the production of antibiotics by plate assay and then cultured in shake flask fermentation at 30ºC for further studies. The bioactive secondary metabolites producing bacterial isolates were studied for their ability to tolerate 3% NaCl. Identification of Bacillus pumilus strains was done by using biochemical test as well as 16S r-RNA sequencing method. Identification of antibiotics was done by column chromatography as well as thin layer chromatography. Antibiotics was found to be produced by three are bacterial and one are fungal pathogen strains against E-coli (ATCC#2939), Staphylococcus aureus (ATCC# 96), Pseudomonas aeruginosa (ATCC# 2488) and one are fungal pathogen strains against Candida albicans (ATCC# 227) proved to be resistant to antibiotics produced by Bacillus pumilus. The maximum production of antibiotics from Bacillus pumilus against E-coli, Staphylococcus aureus and Candida albicans. Maximum zones of inhibition were observed after 48 hours of incubation at 30ºC against E-coli, Staphylococcus aureus and Candida albicans. After that structural elucidation was done by using IR, MS and NMR spectroscopy respectively.

Screening and characterization of protease-producing Virgibacillus, Halobacillus and Oceanobacillus strains from Thai fermented fish.

Halophilic bacteria from fermented fish (pla-ra and pla-chom), shrimp paste (ka-pi) and fermented crab] were isolated and screened for protease activities. Twenty-seven isolates exhibited protease specific activities ranged from 0.12 to 3.62 unit/mg protein. Isolate TPPN2-1 from pla-ra produced maximal protease specific activity (3.62 units/mg protein). They were Gram-positive, moderately halophilic rod-shaped bacteria and were divided into 5 groups on the basis of their phenotypic and chemotaxonomic characteristics, DNA-DNA relatedness including the 16S rRNA gene sequences analyses. Three isolates (Group I) were identified as Virgibacillus halodenitrificans, 3 isolates (Group II) as V. marismortui, 7 isolates (Group III) as V. dokdonensis, 10 isolates (Group IVA & IVB) as Halobacillus species and 4 isolates (Group V) as Oceanobacillus iheyensis.

Survey on Halophilic microbial diversity of Kovalam Saltpans in Kanyakumari District and its industrial applications.

Halophilic microorganisms are organisms that grow optimally in the presence of NaCl at least 0.2 M. The applications of halophilic bacteria include food and pharmaceutical industries, production of enzymes, polymers and various cosmetic products. The objective of the study was isolation and its characterization of potentially important microorganisms from saltpans. In this preliminary investigation, the total microbial counts were studied from the salt sample at three month intervals for one year. Totally 9 organisms were identified based on standard cultural, physiological and biochemical studies. These strains were subjected to screening of potential enzymes such as; amylase, protease, gelatinase, etc. The organisms Natranobacterium sp-1, Staphylococcus epidermidis, Staphylococcus intermedius and Staphylococcus citreus showed positive for amylase; Halo bacillus salinus, Halococcus salifodinae, Staphylococcus epidermidis and Staphylococcus intermedius showed positive for protease; Halobacillus salinus, Halobacterium salinarum, Natranobacterium sp-1, Staphylococcus intermedius and Staphylococcus citreus showed positive for gelatinase activity.

Polymorphic Halophilic Bacteria and Haloarchaea from Lianyungang Taibei and Yancheng Sanwei Salt Field Soils / 微生物学通报

Halophilic microorganisms play important rules in salt field ecosystem and salt production. In this study, halophilic bacteria and haloarchaea from soils of Lianyungang Taibei and Yancheng Sanwei salt fields were analyzed. The halophilic bacterial and haloarchaeal types from both the soils were similar, but each soil had its distinctive species. A total of 17 halophilic bacteria were identified, among them, Halomonas was found from both the soils, while Pontibacillus and Halobacillus were isolated from Sanwei salt field only. Using uncultured 16S rRNA gene library technology, 13 haloarchaeal soil 16S rRNA genes were identified from both the saline soils. Halobacterium and Haloplanus were found from Taibei salt field, while Halobac- terium, Natronobacterium, Halogeometricum and Haloarcula were identified from Sanwei salt field. Ten haloarchaeal 16S rRNA gene sequences showed 92%~97% identities with the GenBank sequences that ap- pear to represent novel soil haloarchaeal species. This study provides important information that is useful for further investigation and application of halophiles of saline soil fields.

Isolation and 16S rRNA Gene Sequence Analysis of a Thermotolerant Halophilic Bacterium / 微生物学通报

A strain of thermotolerant halophilic bacteria YJ0238 was isolated from the salt well at 47℃ in the Kangning countryside where was located at the side of Lancang River in the Tibet Autonomous Region. Some physiological and biochemical properties were characterized.16S rRNA gene sequence of YJ0238 was amplified by PCR,and its nucleotide sequence was determined. Based on it’s physiological and biochemical properties,homology and phylogenetic analysis of 16S rRNA gene sequence,strain YJ0238 was identified as a subspecies of the species Idiomarina zobellii. The GenBank accession number of the 16S rRNA gene sequence of strain YJ0238 is EF693953. Until now,there were few reports on the study of high-temperature and high-salt microbial in domestics. The results of this study will provide research material and information for further studies in this area.

Osmoregulation of a Halophilic Bacteria Strain Halomonas sp.BYS-1 / 微生物学通报

Halomonas sp.BYS-1 was a moderately halophilic bacteria strain isolated from activated sludge,It could grow on MM with NaCl concentration from 0.1～2.6 mol/L and phenyl acetic acid as sole carbon souce. When BYS-1 grew in the media with different concentrations of NaCl, there was no obvious change in its intracellular Na+ contents , it accumulated K+, glutamic acid and betaine as osmoprotectants. Its intracellular contents of K+,glutamic acid and betaine increased by 1.9,2.4 and 13.6 times, respectively, when the concentration of NaCl increased from 0.1 mol/L to 2mol/L.

NUMERICAL TAXONOMY OF MODERATELY HALOPHILIC BACTERIA ISOLATED FROM SALT LAKES IN XINJIANG REGION / 微生物学通报

Twenty nine moderately halophilic bacteria isolated from salt lakes in Xinjiang Region were compared with nine reference strains of relative species by performing a numerical analysis of 112 phenotypic characteristics. All the new isolates are Gram negative and can grow on the media containing 0～25% NaCl. The results showed that all tested strains were clustered into 2 phena at 76 % similarity level. Phenon I was composed by 25 new isolates strains,and phenon II included 9 reference strains and 4 new lsolated strains. Most of the new isolated strains were clusted into 3 subphena at 86 % similarity level. There were 10 new isolated strains in subphenon 1, 8 new isolated strains in subphenon 2, and 1 new isolated strain and 5 reference strains were included in subphenon 3. The results indicated that there were great differences between the new isolates and the reference strains.

16S rDNA PCR-RFLP ANALYSIS OF MODERATELY HALOPHILIC BACTERIA ISOLATED FROM SALT LAKES IN XINJIANG REGION / 微生物学通报

Twenty-eight moderately halophilic bacteria isolated from salt lakes in Xinjiang Regions were analyzed by 16S rDNA PCR-RFLP in comparison with nine reference strains of related species. The 16S rRNA PCR products of the tested stains were digested by four restriction enzymes (AluI, HinfI, RsaI and HaeⅢ). The results showed that all tested strains were clustered into three groups at 74% similarity level. Group I included 2 reference strains Chromohalobacter marismortui, Nesterenkonia halobia and new isolate CI. Group Ⅱ included seven type strains belonging to Halomonas and eight new isolates. Group Ⅲ included ninteen new isolates.