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Katsumi Matsuura - One of the best experts on this subject based on the ideXlab platform.
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Desulfovibrio marinisediminis sp. nov., a novel sulfate-reducing bacterium isolated from coastal marine sediment via enrichment with Casamino Acids.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2008Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Yumiko Hase, Yutaka Uyeno, Katsumi MatsuuraAbstract:To obtain amino acid-utilizing sulfate reducers, enrichment culture was carried out with a medium containing Casamino Acids and sulfate and inoculated with coastal marine sediment from the eutrophic Tokyo Bay, Japan. A sulfate reducer, designated strain C/L2T, was isolated from the sulfide-producing enrichment culture after further enrichment with lactate and sulfate by means of the agar shake dilution method. Cells of strain C/L2T were vibrio-shaped, Gram-negative, motile rods (0.7–1.0 μm wide and 1.0–3.5 μm long) with single polar flagella. The optimum temperature for its growth was 37 °C, the optimum pH was around 7.5 and the optimum NaCl concentration was 20–25 g l−1. Hydrogen, formate, lactate, pyruvate, fumarate, malate, succinate, ethanol, propanol, glycerol, glycine, alanine, serine, aspartate, Casamino Acids, peptone and yeast extract were used as electron donors. Sulfate, sulfite and thiosulfate each served as an electron acceptor, but elemental sulfur, nitrate, fumarate, acrylate and 2,4,6-tribromophenol did not. Disproportionation of thiosulfate was not observed. Desulfoviridin, c-type cytochromes and catalase were present. The major respiratory quinone was MK-6(H2). The G+C content of the genomic DNA was 46.2 mol%. Comparisons based on 16S rRNA gene sequences and on dissimilatory sulfite reductase gene sequences clearly showed that strain C/L2T belonged to the genus Desulfovibrio: its closest relatives were the uncharacterized Desulfovibrio sp. strain TBP-1 (16S rRNA gene sequence similarity of 99.4 %) and Desulfovibrio acrylicus DSM 10141T (16S rRNA gene sequence similarity of 98.7 %). The level of DNA–DNA hybridization with Desulfovibrio acrylicus DSM 10141T was 10.3 %. On the basis of the data from this study and the physiological and phylogenetic differences that exist between the isolate and Desulfovibrio acrylicus, strain C/L2T represents a novel species of the genus Desulfovibrio, for which the name Desulfovibrio marinisediminis sp. nov. is proposed. The type strain is C/L2T (=NMRC 101113T=JCM 14577T=DSM 17456T).
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Desulfovibrio marinisediminis sp. nov., a novel sulfate-reducing bacterium isolated from coastal marine sediment via enrichment with Casamino Acids.
International journal of systematic and evolutionary microbiology, 2008Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Yumiko Hase, Yutaka Uyeno, Katsumi MatsuuraAbstract:To obtain amino acid-utilizing sulfate reducers, enrichment culture was carried out with a medium containing Casamino Acids and sulfate and inoculated with coastal marine sediment from the eutrophic Tokyo Bay, Japan. A sulfate reducer, designated strain C/L2(T), was isolated from the sulfide-producing enrichment culture after further enrichment with lactate and sulfate by means of the agar shake dilution method. Cells of strain C/L2(T) were vibrio-shaped, Gram-negative, motile rods (0.7-1.0 mum wide and 1.0-3.5 mum long) with single polar flagella. The optimum temperature for its growth was 37 degrees C, the optimum pH was around 7.5 and the optimum NaCl concentration was 20-25 g l(-1). Hydrogen, formate, lactate, pyruvate, fumarate, malate, succinate, ethanol, propanol, glycerol, glycine, alanine, serine, aspartate, Casamino Acids, peptone and yeast extract were used as electron donors. Sulfate, sulfite and thiosulfate each served as an electron acceptor, but elemental sulfur, nitrate, fumarate, acrylate and 2,4,6-tribromophenol did not. Disproportionation of thiosulfate was not observed. Desulfoviridin, c-type cytochromes and catalase were present. The major respiratory quinone was MK-6(H(2)). The G+C content of the genomic DNA was 46.2 mol%. Comparisons based on 16S rRNA gene sequences and on dissimilatory sulfite reductase gene sequences clearly showed that strain C/L2(T) belonged to the genus Desulfovibrio: its closest relatives were the uncharacterized Desulfovibrio sp. strain TBP-1 (16S rRNA gene sequence similarity of 99.4 %) and Desulfovibrio acrylicus DSM 10141(T) (16S rRNA gene sequence similarity of 98.7 %). The level of DNA-DNA hybridization with Desulfovibrio acrylicus DSM 10141(T) was 10.3 %. On the basis of the data from this study and the physiological and phylogenetic differences that exist between the isolate and Desulfovibrio acrylicus, strain C/L2(T) represents a novel species of the genus Desulfovibrio, for which the name Desulfovibrio marinisediminis sp. nov. is proposed. The type strain is C/L2(T) (=NBRC [corrected] 101113(T)=JCM 14577(T)=DSM 17456(T)).
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Dethiosulfatibacter aminovorans gen. nov., sp. nov., a novel thiosulfate-reducing bacterium isolated from coastal marine sediment via sulfate-reducing enrichment with Casamino Acids.
International Journal of Systematic and Evolutionary Microbiology, 2007Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Ueno Yutaka, Yuji Sekiguchi, Akihiro Ibe, Katsumi MatsuuraAbstract:A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino Acids as a substrate. A thiosulfate reducer, strain C/G2T, was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2T were non-motile rods (0.6–0.8 μm×2.2–4.8 μm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2T stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28–30 °C, the optimum pH was around 7.8 and the optimum salt concentration was 20–30 g l−1. Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino Acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino Acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2T fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2T on Casamino Acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino Acids. The DNA G+C content of strain C/G2T was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2T formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2T (=JCM 13356T=NBRC 101112T=DSM 17477T) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov.
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Dethiosulfatibacter aminovorans gen. nov., sp. nov., a novel thiosulfate-reducing bacterium isolated from coastal marine sediment via sulfate-reducing enrichment with Casamino Acids.
International journal of systematic and evolutionary microbiology, 2007Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Akihiro Ibe, Yutaka Ueno, Katsumi MatsuuraAbstract:A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino Acids as a substrate. A thiosulfate reducer, strain C/G2(T), was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2(T) were non-motile rods (0.6-0.8 microm x 2.2-4.8 microm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2(T) stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28-30 degrees C, the optimum pH was around 7.8 and the optimum salt concentration was 20-30 g l(-1). Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino Acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino Acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2(T) fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2(T) on Casamino Acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino Acids. The DNA G+C content of strain C/G2(T) was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2(T) formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2(T) (=JCM 13356(T)=NBRC 101112(T)=DSM 17477(T)) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov.
Gaurav Batra - One of the best experts on this subject based on the ideXlab platform.
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Casamino Acids facilitate the secretion of recombinant dengue virus serotype-3 envelope domain III in Pichia pastoris
BMC biotechnology, 2016Co-Authors: Neha Kaushik, Deepak Rohila, Upasana Arora, Rajendra Raut, Urpo Lamminmäki, Navin Khanna, Gaurav BatraAbstract:Dengue is a viral disease spread to humans by mosquitoes. Notably, there are four serotypes of dengue viruses (DENV) that places ~40 % of the global population at risk of infection. However, lack of a suitable drug or a preventive vaccine exacerbates the matter further. Envelope domain-III (EDIII) antigen of dengue virus (DENV) has garnered much attention as a promising vaccine candidate for dengue, in addition to its use as a diagnostic intermediate. Hence developing a method for efficient production of high quality recombinant EDIII is important for research and industrial purpose. In this work, a Pichia pastoris system was optimized for the secretory over-expression of DENV serotype-3 EDIII under the control of methanol inducible AOX1 promoter. Temperature alone had a significant impact upon the amount of secretory EDIII, with 2.5-fold increase upon reducing the induction temperature from 30 to 20 °C. However surprisingly, supplementation of culture media with Casamino Acids (CA), further augmented secretory EDIII titer, with a concomitant drop of intracellular EDIII levels at both temperatures. Though, reduction in intracellular retention of EDIII was more prominent at 20 °C than 30 °C. This suggests that CA supplementation facilitates overexpressing P. pastoris cells to secrete more EDIII by reducing the proportion retained intracellularly. Moreover, a bell-shaped correlation was observed between CA concentration and secretory EDIII titer. The maximum EDIII expression level of 187 mg/L was achieved under shake flask conditions with induction at 20 °C in the presence of 1 % CA. The overall increase in EDIII titer was ~9-fold compared to un-optimized conditions. Notably, mouse immune-sera, generated using this purified EDIII antigen, efficiently neutralized the DENV. The strategy described herein could enable fulfilling the mounting demand for recombinant EDIII as well as lay direction to future studies on secretory expression of recombinant proteins in P. pastoris with CA as a media supplement.
Susumu Takii - One of the best experts on this subject based on the ideXlab platform.
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Desulfovibrio marinisediminis sp. nov., a novel sulfate-reducing bacterium isolated from coastal marine sediment via enrichment with Casamino Acids.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2008Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Yumiko Hase, Yutaka Uyeno, Katsumi MatsuuraAbstract:To obtain amino acid-utilizing sulfate reducers, enrichment culture was carried out with a medium containing Casamino Acids and sulfate and inoculated with coastal marine sediment from the eutrophic Tokyo Bay, Japan. A sulfate reducer, designated strain C/L2T, was isolated from the sulfide-producing enrichment culture after further enrichment with lactate and sulfate by means of the agar shake dilution method. Cells of strain C/L2T were vibrio-shaped, Gram-negative, motile rods (0.7–1.0 μm wide and 1.0–3.5 μm long) with single polar flagella. The optimum temperature for its growth was 37 °C, the optimum pH was around 7.5 and the optimum NaCl concentration was 20–25 g l−1. Hydrogen, formate, lactate, pyruvate, fumarate, malate, succinate, ethanol, propanol, glycerol, glycine, alanine, serine, aspartate, Casamino Acids, peptone and yeast extract were used as electron donors. Sulfate, sulfite and thiosulfate each served as an electron acceptor, but elemental sulfur, nitrate, fumarate, acrylate and 2,4,6-tribromophenol did not. Disproportionation of thiosulfate was not observed. Desulfoviridin, c-type cytochromes and catalase were present. The major respiratory quinone was MK-6(H2). The G+C content of the genomic DNA was 46.2 mol%. Comparisons based on 16S rRNA gene sequences and on dissimilatory sulfite reductase gene sequences clearly showed that strain C/L2T belonged to the genus Desulfovibrio: its closest relatives were the uncharacterized Desulfovibrio sp. strain TBP-1 (16S rRNA gene sequence similarity of 99.4 %) and Desulfovibrio acrylicus DSM 10141T (16S rRNA gene sequence similarity of 98.7 %). The level of DNA–DNA hybridization with Desulfovibrio acrylicus DSM 10141T was 10.3 %. On the basis of the data from this study and the physiological and phylogenetic differences that exist between the isolate and Desulfovibrio acrylicus, strain C/L2T represents a novel species of the genus Desulfovibrio, for which the name Desulfovibrio marinisediminis sp. nov. is proposed. The type strain is C/L2T (=NMRC 101113T=JCM 14577T=DSM 17456T).
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Desulfovibrio marinisediminis sp. nov., a novel sulfate-reducing bacterium isolated from coastal marine sediment via enrichment with Casamino Acids.
International journal of systematic and evolutionary microbiology, 2008Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Yumiko Hase, Yutaka Uyeno, Katsumi MatsuuraAbstract:To obtain amino acid-utilizing sulfate reducers, enrichment culture was carried out with a medium containing Casamino Acids and sulfate and inoculated with coastal marine sediment from the eutrophic Tokyo Bay, Japan. A sulfate reducer, designated strain C/L2(T), was isolated from the sulfide-producing enrichment culture after further enrichment with lactate and sulfate by means of the agar shake dilution method. Cells of strain C/L2(T) were vibrio-shaped, Gram-negative, motile rods (0.7-1.0 mum wide and 1.0-3.5 mum long) with single polar flagella. The optimum temperature for its growth was 37 degrees C, the optimum pH was around 7.5 and the optimum NaCl concentration was 20-25 g l(-1). Hydrogen, formate, lactate, pyruvate, fumarate, malate, succinate, ethanol, propanol, glycerol, glycine, alanine, serine, aspartate, Casamino Acids, peptone and yeast extract were used as electron donors. Sulfate, sulfite and thiosulfate each served as an electron acceptor, but elemental sulfur, nitrate, fumarate, acrylate and 2,4,6-tribromophenol did not. Disproportionation of thiosulfate was not observed. Desulfoviridin, c-type cytochromes and catalase were present. The major respiratory quinone was MK-6(H(2)). The G+C content of the genomic DNA was 46.2 mol%. Comparisons based on 16S rRNA gene sequences and on dissimilatory sulfite reductase gene sequences clearly showed that strain C/L2(T) belonged to the genus Desulfovibrio: its closest relatives were the uncharacterized Desulfovibrio sp. strain TBP-1 (16S rRNA gene sequence similarity of 99.4 %) and Desulfovibrio acrylicus DSM 10141(T) (16S rRNA gene sequence similarity of 98.7 %). The level of DNA-DNA hybridization with Desulfovibrio acrylicus DSM 10141(T) was 10.3 %. On the basis of the data from this study and the physiological and phylogenetic differences that exist between the isolate and Desulfovibrio acrylicus, strain C/L2(T) represents a novel species of the genus Desulfovibrio, for which the name Desulfovibrio marinisediminis sp. nov. is proposed. The type strain is C/L2(T) (=NBRC [corrected] 101113(T)=JCM 14577(T)=DSM 17456(T)).
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Dethiosulfatibacter aminovorans gen. nov., sp. nov., a novel thiosulfate-reducing bacterium isolated from coastal marine sediment via sulfate-reducing enrichment with Casamino Acids.
International Journal of Systematic and Evolutionary Microbiology, 2007Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Ueno Yutaka, Yuji Sekiguchi, Akihiro Ibe, Katsumi MatsuuraAbstract:A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino Acids as a substrate. A thiosulfate reducer, strain C/G2T, was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2T were non-motile rods (0.6–0.8 μm×2.2–4.8 μm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2T stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28–30 °C, the optimum pH was around 7.8 and the optimum salt concentration was 20–30 g l−1. Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino Acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino Acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2T fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2T on Casamino Acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino Acids. The DNA G+C content of strain C/G2T was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2T formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2T (=JCM 13356T=NBRC 101112T=DSM 17477T) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov.
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Dethiosulfatibacter aminovorans gen. nov., sp. nov., a novel thiosulfate-reducing bacterium isolated from coastal marine sediment via sulfate-reducing enrichment with Casamino Acids.
International journal of systematic and evolutionary microbiology, 2007Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Akihiro Ibe, Yutaka Ueno, Katsumi MatsuuraAbstract:A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino Acids as a substrate. A thiosulfate reducer, strain C/G2(T), was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2(T) were non-motile rods (0.6-0.8 microm x 2.2-4.8 microm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2(T) stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28-30 degrees C, the optimum pH was around 7.8 and the optimum salt concentration was 20-30 g l(-1). Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino Acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino Acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2(T) fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2(T) on Casamino Acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino Acids. The DNA G+C content of strain C/G2(T) was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2(T) formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2(T) (=JCM 13356(T)=NBRC 101112(T)=DSM 17477(T)) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov.
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Amino Acids as main substrates for sulfate-reducing bacteria in surface sediment of a eutrophic bay.
The Journal of general and applied microbiology, 2003Co-Authors: Susumu TakiiAbstract:The inner part of Tokyo Bay, Japan, is highly eutrophicated as shown by the frequent occurrence of red tide. The bottom water is anoxic during warm seasons especially at artificially dredged sites. In the sediment slurries prepared from surface sediment samples collected from the dredged sites, substrate addition stimulated the consumption of sulfate during anaerobic incubation. Of the substrates added, the seston composed mainly of diatom stimulated consumption more than lactate and acetate. Its effect was nearly equal to that of Casamino Acids. Casamino Acids and some amino Acids also accelerated the rate of sulfate reduction measured by the tracer method in sediment samples more than lactate or acetate. Anaerobic incubation of the sediment slurry amended with Casamino Acids showed that the consumption of amino Acids was retarded by the addition of molybdate (final concentration; 20 mM). In the slurry amended with only molybdate, glutamate was accumulated distinctively and linearly with time. Its accumulation rate in molar base was comparable to the rate of sulfate reduction. These results suggested that amino Acids were the main substrates for sulfate-reducing bacteria (SRB) in the sediment. The MPN values of SRB in these sediment samples were often higher with the enumeration medium containing Casamino Acids instead of lactate. Furthermore, during a week incubation of sediment slurries amended with substrates, Casamino Acids and seston more greatly stimulated the growth of SRB enumerated by both media than lactate.
Satoshi Hanada - One of the best experts on this subject based on the ideXlab platform.
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Desulfovibrio marinisediminis sp. nov., a novel sulfate-reducing bacterium isolated from coastal marine sediment via enrichment with Casamino Acids.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2008Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Yumiko Hase, Yutaka Uyeno, Katsumi MatsuuraAbstract:To obtain amino acid-utilizing sulfate reducers, enrichment culture was carried out with a medium containing Casamino Acids and sulfate and inoculated with coastal marine sediment from the eutrophic Tokyo Bay, Japan. A sulfate reducer, designated strain C/L2T, was isolated from the sulfide-producing enrichment culture after further enrichment with lactate and sulfate by means of the agar shake dilution method. Cells of strain C/L2T were vibrio-shaped, Gram-negative, motile rods (0.7–1.0 μm wide and 1.0–3.5 μm long) with single polar flagella. The optimum temperature for its growth was 37 °C, the optimum pH was around 7.5 and the optimum NaCl concentration was 20–25 g l−1. Hydrogen, formate, lactate, pyruvate, fumarate, malate, succinate, ethanol, propanol, glycerol, glycine, alanine, serine, aspartate, Casamino Acids, peptone and yeast extract were used as electron donors. Sulfate, sulfite and thiosulfate each served as an electron acceptor, but elemental sulfur, nitrate, fumarate, acrylate and 2,4,6-tribromophenol did not. Disproportionation of thiosulfate was not observed. Desulfoviridin, c-type cytochromes and catalase were present. The major respiratory quinone was MK-6(H2). The G+C content of the genomic DNA was 46.2 mol%. Comparisons based on 16S rRNA gene sequences and on dissimilatory sulfite reductase gene sequences clearly showed that strain C/L2T belonged to the genus Desulfovibrio: its closest relatives were the uncharacterized Desulfovibrio sp. strain TBP-1 (16S rRNA gene sequence similarity of 99.4 %) and Desulfovibrio acrylicus DSM 10141T (16S rRNA gene sequence similarity of 98.7 %). The level of DNA–DNA hybridization with Desulfovibrio acrylicus DSM 10141T was 10.3 %. On the basis of the data from this study and the physiological and phylogenetic differences that exist between the isolate and Desulfovibrio acrylicus, strain C/L2T represents a novel species of the genus Desulfovibrio, for which the name Desulfovibrio marinisediminis sp. nov. is proposed. The type strain is C/L2T (=NMRC 101113T=JCM 14577T=DSM 17456T).
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Desulfovibrio marinisediminis sp. nov., a novel sulfate-reducing bacterium isolated from coastal marine sediment via enrichment with Casamino Acids.
International journal of systematic and evolutionary microbiology, 2008Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Yumiko Hase, Yutaka Uyeno, Katsumi MatsuuraAbstract:To obtain amino acid-utilizing sulfate reducers, enrichment culture was carried out with a medium containing Casamino Acids and sulfate and inoculated with coastal marine sediment from the eutrophic Tokyo Bay, Japan. A sulfate reducer, designated strain C/L2(T), was isolated from the sulfide-producing enrichment culture after further enrichment with lactate and sulfate by means of the agar shake dilution method. Cells of strain C/L2(T) were vibrio-shaped, Gram-negative, motile rods (0.7-1.0 mum wide and 1.0-3.5 mum long) with single polar flagella. The optimum temperature for its growth was 37 degrees C, the optimum pH was around 7.5 and the optimum NaCl concentration was 20-25 g l(-1). Hydrogen, formate, lactate, pyruvate, fumarate, malate, succinate, ethanol, propanol, glycerol, glycine, alanine, serine, aspartate, Casamino Acids, peptone and yeast extract were used as electron donors. Sulfate, sulfite and thiosulfate each served as an electron acceptor, but elemental sulfur, nitrate, fumarate, acrylate and 2,4,6-tribromophenol did not. Disproportionation of thiosulfate was not observed. Desulfoviridin, c-type cytochromes and catalase were present. The major respiratory quinone was MK-6(H(2)). The G+C content of the genomic DNA was 46.2 mol%. Comparisons based on 16S rRNA gene sequences and on dissimilatory sulfite reductase gene sequences clearly showed that strain C/L2(T) belonged to the genus Desulfovibrio: its closest relatives were the uncharacterized Desulfovibrio sp. strain TBP-1 (16S rRNA gene sequence similarity of 99.4 %) and Desulfovibrio acrylicus DSM 10141(T) (16S rRNA gene sequence similarity of 98.7 %). The level of DNA-DNA hybridization with Desulfovibrio acrylicus DSM 10141(T) was 10.3 %. On the basis of the data from this study and the physiological and phylogenetic differences that exist between the isolate and Desulfovibrio acrylicus, strain C/L2(T) represents a novel species of the genus Desulfovibrio, for which the name Desulfovibrio marinisediminis sp. nov. is proposed. The type strain is C/L2(T) (=NBRC [corrected] 101113(T)=JCM 14577(T)=DSM 17456(T)).
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Dethiosulfatibacter aminovorans gen. nov., sp. nov., a novel thiosulfate-reducing bacterium isolated from coastal marine sediment via sulfate-reducing enrichment with Casamino Acids.
International Journal of Systematic and Evolutionary Microbiology, 2007Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Ueno Yutaka, Yuji Sekiguchi, Akihiro Ibe, Katsumi MatsuuraAbstract:A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino Acids as a substrate. A thiosulfate reducer, strain C/G2T, was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2T were non-motile rods (0.6–0.8 μm×2.2–4.8 μm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2T stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28–30 °C, the optimum pH was around 7.8 and the optimum salt concentration was 20–30 g l−1. Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino Acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino Acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2T fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2T on Casamino Acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino Acids. The DNA G+C content of strain C/G2T was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2T formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2T (=JCM 13356T=NBRC 101112T=DSM 17477T) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov.
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Dethiosulfatibacter aminovorans gen. nov., sp. nov., a novel thiosulfate-reducing bacterium isolated from coastal marine sediment via sulfate-reducing enrichment with Casamino Acids.
International journal of systematic and evolutionary microbiology, 2007Co-Authors: Susumu Takii, Satoshi Hanada, Hideyuki Tamaki, Yuji Sekiguchi, Akihiro Ibe, Yutaka Ueno, Katsumi MatsuuraAbstract:A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino Acids as a substrate. A thiosulfate reducer, strain C/G2(T), was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2(T) were non-motile rods (0.6-0.8 microm x 2.2-4.8 microm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2(T) stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28-30 degrees C, the optimum pH was around 7.8 and the optimum salt concentration was 20-30 g l(-1). Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino Acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino Acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2(T) fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2(T) on Casamino Acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino Acids. The DNA G+C content of strain C/G2(T) was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2(T) formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2(T) (=JCM 13356(T)=NBRC 101112(T)=DSM 17477(T)) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov.
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Photosynthetic Apparatus in Roseateles depolymerans 61A Is Transcriptionally Induced by Carbon Limitation
Applied and environmental microbiology, 2002Co-Authors: Tetsushi Suyama, Toru Shigematsu, Toshihiko Suzuki, Yutaka Tokiwa, Takahiro Kanagawa, Kenji V. P. Nagashima, Satoshi HanadaAbstract:Production of a photosynthetic apparatus in Roseateles depolymerans 61A, a recently discovered freshwater β-Proteobacterium showing characteristics of aerobic phototrophic bacteria, was observed when the cells were subjected to a sudden decrease in carbon sources (e.g., when cells grown with 0.1 to 0.4% Casamino Acids were diluted or transferred into medium containing ≤0.04% Casamino Acids). Accumulation of bacteriochlorophyll (BChl) a was observed in the presence of oxygen and was enhanced under semiaerobic conditions (2% oxygen) but was reduced in the presence of light. Similarly to what has been reported regarding some aerobic phototrophic bacteria belonging to the α subclass of the Proteobacteria, viability of the cells in the carbon source-free medium was prolonged under aerobic-light (10 W m−2) conditions, possibly due to photosynthetic energy conversion, but was not prolonged under aerobic-dark conditions. The puf operon, which encodes most of the apoproteins of light-harvesting and reaction center complexes, was sequenced, and the effect of changes in Casamino Acids concentrations, oxygen, and light on its expression was estimated by the accumulation of its mRNA. The expression of the puf operon was induced by the decrease in carbon sources, similarly to what was observed for the accumulation of BChl a under aerobic and semiaerobic conditions (≥0.2% O2), and was reduced in the presence of light. Transcription of the R. depolymerans puf operon is considered to be controlled by changes in carbon nutrients in addition to oxygen tension and light intensity.
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Casamino Acids facilitate the secretion of recombinant dengue virus serotype-3 envelope domain III in Pichia pastoris
BMC biotechnology, 2016Co-Authors: Neha Kaushik, Deepak Rohila, Upasana Arora, Rajendra Raut, Urpo Lamminmäki, Navin Khanna, Gaurav BatraAbstract:Dengue is a viral disease spread to humans by mosquitoes. Notably, there are four serotypes of dengue viruses (DENV) that places ~40 % of the global population at risk of infection. However, lack of a suitable drug or a preventive vaccine exacerbates the matter further. Envelope domain-III (EDIII) antigen of dengue virus (DENV) has garnered much attention as a promising vaccine candidate for dengue, in addition to its use as a diagnostic intermediate. Hence developing a method for efficient production of high quality recombinant EDIII is important for research and industrial purpose. In this work, a Pichia pastoris system was optimized for the secretory over-expression of DENV serotype-3 EDIII under the control of methanol inducible AOX1 promoter. Temperature alone had a significant impact upon the amount of secretory EDIII, with 2.5-fold increase upon reducing the induction temperature from 30 to 20 °C. However surprisingly, supplementation of culture media with Casamino Acids (CA), further augmented secretory EDIII titer, with a concomitant drop of intracellular EDIII levels at both temperatures. Though, reduction in intracellular retention of EDIII was more prominent at 20 °C than 30 °C. This suggests that CA supplementation facilitates overexpressing P. pastoris cells to secrete more EDIII by reducing the proportion retained intracellularly. Moreover, a bell-shaped correlation was observed between CA concentration and secretory EDIII titer. The maximum EDIII expression level of 187 mg/L was achieved under shake flask conditions with induction at 20 °C in the presence of 1 % CA. The overall increase in EDIII titer was ~9-fold compared to un-optimized conditions. Notably, mouse immune-sera, generated using this purified EDIII antigen, efficiently neutralized the DENV. The strategy described herein could enable fulfilling the mounting demand for recombinant EDIII as well as lay direction to future studies on secretory expression of recombinant proteins in P. pastoris with CA as a media supplement.