scholarly journals Thermostable Chitosanase from Bacillus sp. Strain CK4: Cloning and Expression of the Gene and Characterization of the Enzyme

2000 ◽  
Vol 66 (9) ◽  
pp. 3727-3734 ◽  
Author(s):  
Ho-Geun Yoon ◽  
Hee-Yun Kim ◽  
Young-Hee Lim ◽  
Hye-Kyung Kim ◽  
Dong-Hoon Shin ◽  
...  
Keyword(s):  
Gene Sequence ◽  
Specific Activity ◽  
High Specific Activity ◽  
Industrial Applications ◽  
Cloning And Expression ◽  
Rrna Gene ◽  
Phenotypic Analysis ◽  
Reading Frame ◽  
The 16S Rrna Gene

ABSTRACT A thermostable chitosanase gene from the environmental isolateBacillus sp. strain CK4, which was identified on the basis of phylogenetic analysis of the 16S rRNA gene sequence and phenotypic analysis, was cloned, and its complete DNA sequence was determined. The thermostable chitosanase gene was composed of an 822-bp open reading frame which encodes a protein of 242 amino acids and a signal peptide corresponding to a 30-kDa enzyme. The deduced amino acid sequence of the chitosanase from Bacillus sp. strain CK4 exhibits 76.6, 15.3, and 14.2% similarities to those from Bacillus subtilis, Bacillus ehemensis, and Bacillus circulans, respectively. C-terminal homology analysis shows thatBacillus sp. strain CK4 belongs to cluster III withB. subtilis. The gene was similar in size to that of the mesophile B. subtilis but showed a higher preference for codons ending in G or C. The enzyme contains 2 additional cysteine residues at positions 49 and 211. The recombinant chitosanase has been purified to homogeneity by using only two steps with column chromatography. The half-life of the enzyme was 90 min at 80�C, which indicates its usefulness for industrial applications. The enzyme had a useful reactivity and a high specific activity for producing functional oligosaccharides as well, with trimers through hexamers as the major products.

scholarly journals Cloning and Expression of the algL Gene, Encoding the Azotobacter chroococcum Alginate Lyase: Purification and Characterization of the Enzyme

1999 ◽  
Vol 181 (5) ◽  
pp. 1409-1414 ◽  
Author(s):  
Ana Peciña ◽  
Alberto Pascual ◽  
Antonio Paneque
Keyword(s):  
Gene Sequence ◽  
Azotobacter Vinelandii ◽  
Alginate Lyase ◽  
Azotobacter Chroococcum ◽  
Open Reading Frame ◽  
Cloning And Expression ◽  
Gene Encoding ◽  
Reading Frame ◽  
Encoding Gene

ABSTRACT The alginate lyase-encoding gene (algL) ofAzotobacter chroococcum was localized to a 3.1-kbEcoRI DNA fragment that revealed an open reading frame of 1,116 bp. This open reading frame encodes a protein of 42.98 kDa, in agreement with the value previously reported by us for this protein. The deduced protein has a potential N-terminal signal peptide that is consistent with its proposed periplasmic location. The analysis of the deduced amino acid sequence indicated that the gene sequence has a high homology (90% identity) to the Azotobacter vinelandii gene sequence, which has very recently been deposited in the GenBank database, and that it has 64% identity to the Pseudomonas aeruginosa gene sequence but that it has rather low homology (15 to 22% identity) to the gene sequences encoding alginate lyase in other bacteria. The A. chroococcum AlgL protein was overproduced in Escherichia coli and purified to electrophoretic homogeneity in a two-step chromatography procedure on hydroxyapatite and phenyl-Sepharose. The kinetic and molecular parameters of the recombinant alginate lyase are similar to those found for the native enzyme.

scholarly journals Aeromonas fluvialis sp. nov., isolated from a Spanish river

2010 ◽  
Vol 60 (1) ◽  
pp. 72-77 ◽  
Author(s):  
Anabel Alperi ◽  
Antonio J. Martínez-Murcia ◽  
Arturo Monera ◽  
Maria J. Saavedra ◽  
Maria J. Figueras
Keyword(s):  
Gene Sequence ◽  
Housekeeping Genes ◽  
Rrna Gene ◽  
Phenotypic Analysis ◽  
North East ◽  
Aeromonas Veronii ◽  
Aeromonas Sobria ◽  
Dna Reassociation ◽  
Species Being ◽  
The 16S Rrna Gene

A Gram-stain-negative, facultatively anaerobic bacterial strain, designated 717T, was isolated from a water sample collected from the Muga river, Girona, north-east Spain. Preliminary analysis of the 16S rRNA gene sequence showed that this strain belonged to the genus Aeromonas, the nearest species being Aeromonas veronii (99.5 % similarity, with seven different nucleotides). A polyphasic study based on a multilocus phylogenetic analysis of five housekeeping genes (gyrB, rpoD, recA, dnaJ and gyrA; 3684 bp) showed isolate 717T to be an independent phylogenetic line, with Aeromonas sobria, Aeromonas veronii and Aeromonas allosaccharophila as the closest neighbour species. DNA–DNA reassociation experiments and phenotypic analysis identified that strain 717T represents a novel species, for which the name Aeromonas fluvialis sp. nov. is proposed, with type strain 717T (=CECT 7401T =LMG 24681T).

scholarly journals Characterization of uncultured giant rod-shaped magnetotactic Gammaproteobacteria from a freshwater pond in Kanazawa, Japan

Microbiology ◽  
2014 ◽  
Vol 160 (10) ◽  
pp. 2226-2234 ◽  
Author(s):  
Azuma Taoka ◽  
Junya Kondo ◽  
Zachery Oestreicher ◽  
Yoshihiro Fukumori
Keyword(s):  
New Genus ◽  
Gene Sequence ◽  
Rrna Gene ◽  
Comprehensive Understanding ◽  
Dense Granules ◽  
Freshwater Pond ◽  
Bacterium Strain ◽  
And Function ◽  
The 16S Rrna Gene

Magnetotactic bacteria (MTB) are widespread aquatic bacteria, and are a phylogenetically, physiologically and morphologically heterogeneous group, but they all have the ability to orientate and move along the geomagnetic field using intracellular magnetic organelles called magnetosomes. Isolation and cultivation of novel MTB are necessary for a comprehensive understanding of magnetosome formation and function in divergent MTB. In this study, we enriched a giant rod-shaped magnetotactic bacterium (strain GRS-1) from a freshwater pond in Kanazawa, Japan. Cells of strain GRS-1 were unusually large (~13×~8 µm). They swam in a helical trajectory towards the south pole of a bar magnet by means of a polar bundle of flagella. Another striking feature of GRS-1 was the presence of two distinct intracellular biomineralized structures: large electron-dense granules composed of calcium and long chains of magnetosomes that surround the large calcium granules. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that this strain belongs to the Gammaproteobacteria and represents a new genus of MTB.

scholarly journals Streptomyces hainanensis sp. nov., a novel member of the genus Streptomyces

2007 ◽  
Vol 57 (11) ◽  
pp. 2694-2698 ◽  
Author(s):  
Yi Jiang ◽  
Shu-Kun Tang ◽  
Jutta Wiese ◽  
Li-Hua Xu ◽  
Johannes F. Imhoff ◽  
...  
Keyword(s):  
Gene Sequence ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Type Ii ◽  
Rrna Gene Sequence ◽  
Genus Streptomyces ◽  
Type Strains ◽  
The 16S Rrna Gene ◽  
Sequence Similarities

A novel isolate belonging to the genus Streptomyces, strain YIM 47672T, was obtained from soil from Hainan, an island in China. The characterization of this isolate was performed by using a polyphasic approach. The strain formed long spore chains in the aerial mycelia. The cell wall contained l-diaminopimelic acid, traces of meso-diaminopimelic acid and glycine. Whole-cell hydrolysates contained galactose and xylose. The phospholipid was of type II. The 16S rRNA gene sequence similarities for YIM 47672T with respect to the most closely related type strains of species of the genus Streptomyces were less than 96.3 %. Therefore strain YIM 47672T represents a novel member of the genus Streptomyces, for which the name Streptomyces hainanensis sp. nov. is proposed. The type strain is YIM 47672T (=CCTCC AA 205017T=DSM 41900T).

scholarly journals Molecular Characterization of BrucellaStrains Isolated from Marine Mammals

2000 ◽  
Vol 38 (3) ◽  
pp. 1258-1262 ◽  
Author(s):  
Betsy J. Bricker ◽  
Darla R. Ewalt ◽  
Alastair P. MacMillan ◽  
Geoff Foster ◽  
Simon Brew
Keyword(s):  
Host Species ◽  
Marine Mammals ◽  
Rrna Gene ◽  
Gram Negative Bacteria ◽  
Phenotypic Analysis ◽  
New Host ◽  
Homologous Sequences ◽  
The 16S Rrna Gene ◽  
New Host Species

Recently, gram-negative bacteria isolated from a variety of marine mammals have been identified as Brucella species by conventional phenotypic analysis. This study found the 16S rRNA gene from one representative isolate was identical to the homologous sequences of Brucella abortus, B. melitensis,B. canis, and B. suis. IS711-based DNA fingerprinting of 23 isolates from marine mammals showed all the isolates differed from the classical Brucella species. In general, fingerprint patterns grouped by host species. The data suggest that the marine mammal isolates are distinct types ofBrucella and not one of the classical species or biovars invading new host species. In keeping with historical precedent, the designation of several new Brucella species may be appropriate.

Isolation of Bacillus megaterium MPF-906 and its Fermentation Characterization of Reducing Nitrite

2011 ◽  
Vol 365 ◽  
pp. 272-279 ◽  
Author(s):  
Ying Zhang ◽  
Ping Liu ◽  
Peng Fei Ma ◽  
Jun She Sun
Keyword(s):  
Aqueous Solution ◽  
Bacillus Megaterium ◽  
Gene Sequence ◽  
Rrna Gene ◽  
Rrna Gene Sequence ◽  
Nitrite Concentration ◽  
Initial Ph ◽  
Physiological Tests ◽  
The 16S Rrna Gene

Nitrites are highly harmful compounds. They are extremely undesirable in surface and municipal water. The permissible content in natural water is very low and should not exceed 0.01 mg of NO2-/dm3. In this paper a Bacillus isolated from the waste tobacco trimmings was able to reduce above 99.8% of NaNO2 under aerobic conditions. Based on the morphology, physiological tests, the 16S rRNA gene sequence and the phylogenetic characteristics, the strain was identified as Bacillus megaterium. The bacteria grew optimally at 30 °C, initial pH 7.0 and 138 g/l of nitrite concentration. After incubated aerobically under agitation 160 rpm for 24 h, 99.8% of NaNO2 was reduced by B. megaterium MPF-906, this showed a promising biology method for the reduction of nitrites in aqueous solution.

scholarly journals CLONING AND EXPRESSION OF A LIPASE GENE FROM PSEUDOMONAS AERUGINOSA INTO E.coli

2019 ◽  
Vol 50 (3) ◽  
Author(s):  
Auda & Khalifa
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Enzymatic Activity ◽  
Solid Medium ◽  
Specific Activity ◽  
Cloning And Expression ◽  
Tween 20 ◽  
Rrna Gene ◽  
High Fat ◽  
Lipase Gene ◽  
The 16S Rrna Gene

Fifteen local isolates of Pseudomonas were obtained from several sources such as soil, water and some high-fat foods (Meat, olives, coconuts, etc.). The ability of isolates to produce lipase was measured by the size of clear zone on Tween 20 solid medium and by measuring the enzymatic activity and specific activity. Isolate M3 (as named in this study) was found to be the most efficient for the production of the lipase with enzymatic activity reached 56.6 U/ml and specific activity of 305.94 U/mg. This isolate was identified through genetic analysis of the 16S rRNA gene. and it was shown that the isolate M3 belongs to Pseudomonas aeruginosa with 99% similarity.  The DNA of isolate M3 was extracted and lipase gene was amplified through PCR technique, then purified and cloned into E.coli DH5α cells first using pTG19-T plasmid, and expressed in E.coli Bl21 with expression vector pet-28a. The activity of lipase from transformed E.coli Bl21 was 196.6 U/ml and the specific activity 618.2 U/mg.

scholarly journals Rhodanobacter thiooxydans sp. nov., isolated from a biofilm on sulfur particles used in an autotrophic denitrification process

2007 ◽  
Vol 57 (8) ◽  
pp. 1775-1779 ◽  
Author(s):  
Chang Soo Lee ◽  
Kwang Kyu Kim ◽  
Zubair Aslam ◽  
Sung-Taik Lee
Keyword(s):  
Dna Hybridization ◽  
Type Strain ◽  
Gene Sequence ◽  
Novel Species ◽  
Rrna Gene ◽  
Autotrophic Denitrification ◽  
Rrna Gene Sequence ◽  
Phenotypic Analysis ◽  
The 16S Rrna Gene ◽  
Denitrification Process

A novel thiosulfate-oxidizing bacterium, designated strain LCS2T, was isolated from a biofilm on sulfur particles used in an autotrophic denitrification process. The strain was found to comprise Gram-negative, non-motile, non-spore-forming rods that produced yellow-pigmented colonies on R2A agar. The strain contained Q-8 as the major ubiquinone and 17 : 1 iso ω9c, 15 : 0 iso and 17 : 0 iso as the major fatty acids. The G+C content of the genomic DNA was 64.6 mol%. The 16S rRNA gene sequence of strain LCS2T was found to be most similar to that of Rhodanobacter fulvus IAM 15025T (97.4 % similarity). The results of DNA–DNA hybridization and phenotypic analysis showed that strain LCS2T can be distinguished from all known Rhodanobacter species and therefore represents a novel species of the genus, for which the name Rhodanobacter thiooxydans sp. nov. is proposed. The type strain is LCS2T (=DSM 18863T =KCTC 12771T).

Sign in / Sign up

Export Citation Format

Share Document