Isolation and characterization of indigo-reducing bacteria and analysis of microbiota from indigo fermentation suspensions

2021 ◽  
Author(s):  
Kasumi Nakagawa ◽  
Michiki Takeuchi ◽  
Manami Tada ◽  
Momoka Matsunaga ◽  
Masami Kugo ◽  
...  
Keyword(s):  
Indigo Carmine ◽  
Water Soluble ◽  
Assay Method ◽  
Reduction Activity ◽  
Isolation And Characterization ◽  
Alkaline Conditions ◽  
Indigo Reduction ◽  
Reducing Bacteria ◽  
First Time ◽  
Indigo Fermentation

ABSTRACT In natural indigo dyeing, the water-insoluble indigo included in the composted indigo leaves called sukumo is converted to water-soluble leuco-indigo through the reduction activities of microorganisms under alkaline conditions. To understand the relationship between indigo reduction and microorganisms in indigo-fermentation suspensions, we isolated and identified the microorganisms that reduce indigo and analyzed the microbiota in indigo-fermentation suspensions. Indigo-reducing microorganisms, which were not isolated by means of a conventional indigo carmine-reduction assay method, were isolated by using indigo as a direct substrate and further identified and characterized. We succeeded in isolating bacteria closely related to Corynebacterium glutamicum, Chryseomicrobium aureum, Enterococcus sp. for the first time. Anthraquinone was found to be an effective mediator that facilitated the indigo-reduction activity of the isolated strains. On analysis of the microbiota in indigo-fermentation suspensions, the ratio of indigo-reducing bacteria and others was found to be important for maintaining the indigo-reduction activity.

scholarly journals Preparation and Application of Bioshell Calcium Oxide (BiSCaO) Nanoparticle-Dispersions with Bactericidal Activity

Molecules ◽  
2019 ◽  
Vol 24 (18) ◽  
pp. 3415 ◽  
Author(s):  
Sato ◽  
Ishihara ◽  
Nakamura ◽  
Fukuda ◽  
Takayama ◽  
...  
Keyword(s):  
Calcium Oxide ◽  
Pure Water ◽  
Bacterial Flora ◽  
Particle Diameter ◽  
Average Diameter ◽  
Water Soluble ◽  
High Ph ◽  
Reduction Activity ◽  
Nanoparticle Dispersions ◽  
Alkaline Conditions

Scallop-shell powder (SSP) heated at high temperature exhibits high pH and broad antimicrobial activity. Bioshell calcium oxide (BiSCaO) is an SSP composed mainly of calcium oxide. It is poorly water-soluble under alkaline conditions and the generated precipitate can plug spray nozzles. The aim of this study was to establish that BiSCaO dispersion caused no significant CaO loss and plugging of spray nozzles, and to evaluate its deodorization and microbicidal abilities and its ability to reduce the concentrations of NO2− and NO3−. BiSCaO dispersions were prepared by mixing various concentrations of BiSCaO suspension, while phosphate compounds such as Na3PO4, Na2HPO4 or NaH2PO4 and the pH, average diameter, zeta potential, and form of the compounds with cryo-SEM were evaluated. We evaluated deodorization using tainted pork meat and microbicidal efficacy using contaminated suspension with normal bacterial flora. The concentration of NO2− and NO3− after mixing BiSCaO dispersion and pure water containing a high proportion of NO2− and NO3− were measured. BiSCaO dispersion formed with Na2HPO4, whose ratio to BiSCaO was 60%, showed a high pH (>12), a small particle diameter (>181 nm) and was stable for seven days. The BiSCaO dispersion showed higher deodorization and microbicidal activities than SSP-Ca(OH)2, which was mainly composed of Ca(OH)2. BiSCaO, but not SSP-Ca(OH)2, could reduce the concentration of NO2− and NO3− by more than 90% within 15 min. We developed a stable BiSCaO dispersion, and it had high deodorization and microbicidal efficacy. These activities of BiSCaO might result from the high pH caused by CaO hydration and a reduction activity causing active radical species.

Isolation and characterization of cell walls from the mesocarp of mature grape berries (Vitis vinifera)

2020 ◽  
Author(s):  
KJ Nunan ◽  
Ian Sims ◽  
A Bacic ◽  
SP Robinson ◽  
GB Fincher
Keyword(s):  
Vitis Vinifera ◽  
Cell Walls ◽  
Vascular Tissue ◽  
Compositional Analysis ◽  
Water Soluble ◽  
Vitis Vinifera L ◽  
Nylon Mesh ◽  
Cytoplasmic Material ◽  
Isolation And Characterization ◽  
Tissue Homogenates

Cell walls have been isolated from the mesocarp of mature grape (Vitis vinifera L.) berries. Tissue homogenates were suspended in 80% (v/v) ethanol to minimise the loss of water-soluble wall components and wet-sieved on nylon mesh to remove cytoplasmic material. The cell wall fragments retained on the sieve were subsequently treated with buffered phenol at pH 7.0, to inactivate any wall-bound enzymes and to dislodge small amounts of cytoplasmic proteins that adhered to the walls. Finally, the wall preparation was washed with chloroform/methanol (1:1, v/v) to remove lipids and dried by solvent exchange. Scanning electron microscopy showed that the wall preparation was essentially free of vascular tissue and adventitious protein of cytoplasmic origin. Compositional analysis showed that the walls consisted of approximately 90% by weight of polysaccharide and less than 10% protein. The protein component of the walls was shown to be rich in arginine and hydroxyproline residues. Cellulose and polygalacturonans were the major constituents, and each accounted for 30-40% by weight of the polysaccharide component of the walls. Substantial varietal differences were observed in the relative abundance of these two polysaccharides. Xyloglucans constituted approximately 10% of the polysaccharide fraction and the remainder was made up of smaller amounts of mannans, heteroxylans, arabinans and galactans.

Natrophosphate from the Ilímaussaq alkaline complex, South Greenland

2001 ◽  
pp. 139-141
Author(s):  
Ole V. Petersen ◽  
Alexander P. Khomyakov ◽  
Henning Sørensen
Keyword(s):  
Refractive Index ◽  
Kola Peninsula ◽  
Unit Cell Parameter ◽  
Cell Parameter ◽  
Water Soluble ◽  
Type Locality ◽  
Drill Core ◽  
Alkaline Complex ◽  
First Time ◽  
Alkaline Complexes

NOTE: This article was published in a former series of GEUS Bulletin. Please use the original series name when citing this article, for example: Petersen, O. V., Khomyakov, A. P., & Henning. (2001). Natrophosphate from the Ilímaussaq alkaline complex, South Greenland. Geology of Greenland Survey Bulletin, 190, 139-141. https://doi.org/10.34194/ggub.v190.5184 _______________ The rare mineral natrophosphate has been identified for the first time in the Ilímaussaq alkaline complex in a drill core from the Kvanefjeld area. It occurs sparsely in zoned veinlets with cores of natrophosphate and borders of fibrous trona. The natrophosphate is more or less smoky, transparent and unaltered. The refractive index n = 1.448 ± 0.005 is low compared to that given for the material from the type locality, Khibina alkaline complex, Kola Peninsula; the unit cell parameter a = 27.76 ± 0.05 Å is in excellent agreement with that given for the material from the type locality. The veins occur in hyper-agpaitic naujakasite lujavrite; villiaumite is an associated mineral. Only a few water-soluble minerals have so far been found in the Ilímaussaq alkaline complex compared to the wealth of such minerals in the Khibina and Lovozero alkaline complexes. This is possibly at least partly due to lack of necessary precautions during sampling.

Isolation and HPLC Determination of the Chemical Components of Gentianella acuta (Michx.) Hulten

2018 ◽  
Vol 15 (1) ◽  
pp. 21-33
Author(s):  
Ying Wei ◽  
Yongqiao Liu ◽  
Yifan Hele ◽  
Weiwei Sun ◽  
Yang Wang ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
High Speed ◽  
Chemical Constituents ◽  
Folk Medicine ◽  
Gradient Elution ◽  
Chemical Components ◽  
Isolation And Characterization ◽  
Major Chemical ◽  
Main Components ◽  
First Time

Background: Gentianella acuta (Michx.) Hulten is an important type of medicinal plant found in several Chinese provinces. It has been widely used in folk medicine to treat various illnesses. However, there is not enough detailed information about the chemical constituents of this plant or methods for their content determination. Objective: The focus of this work is the isolation and characterization of the major chemical constituents of Gentianella acuta, and developing an analytical method for their determination. Methods: The components of Gentianella acuta were isolated using (1) ethanol extraction and adsorption on macroporous resin. (2) and ethyl acetate extraction and high speed countercurrent chromatography. A HPLC-DAD method was developed using a C18 column and water-acetonitrile as the mobile phase. Based on compound polarities, both isocratic and gradient elution methods were developed. Results: A total of 29 compounds were isolated from this plant, of which 17 compounds were isolated from this genus for the first time. The main components in this plant were found to be xanthones. The HPLC-DAD method was developed and validated for their determination, and found to show good sensitivity and reliability. Conclusion: The results of this work add to the limited body of work available on this important medicinal plant. The findings will be useful for further investigation and development of Gentianella acuta for its valuable medicinal properties.

scholarly journals Isolation and characterization of a new cold-active protease from psychrotrophic bacteria of Western Himalayan glacial soil

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Saleem Farooq ◽  
Ruqeya Nazir ◽  
Shabir Ahmad Ganai ◽  
Bashir Ahmad Ganai
Keyword(s):  
Specific Activity ◽  
Temperature Range ◽  
Psychrotrophic Bacteria ◽  
Isolation And Characterization ◽  
Cold Active ◽  
Active Protease ◽  
Quantitative Screening ◽  
First Time ◽  
Low K

AbstractAs an approach to the exploration of cold-active enzymes, in this study, we isolated a cold-active protease produced by psychrotrophic bacteria from glacial soils of Thajwas Glacier, Himalayas. The isolated strain BO1, identified as Bacillus pumilus, grew well within a temperature range of 4–30 °C. After its qualitative and quantitative screening, the cold-active protease (Apr-BO1) was purified. The Apr-BO1 had a molecular mass of 38 kDa and showed maximum (37.02 U/mg) specific activity at 20 °C, with casein as substrate. It was stable and active between the temperature range of 5–35 °C and pH 6.0–12.0, with an optimum temperature of 20 °C at pH 9.0. The Apr-BO1 had low Km value of 1.0 mg/ml and Vmax 10.0 µmol/ml/min. Moreover, it displayed better tolerance to organic solvents, surfactants, metal ions and reducing agents than most alkaline proteases. The results exhibited that it effectively removed the stains even in a cold wash and could be considered a decent detergent additive. Furthermore, through protein modelling, the structure of this protease was generated from template, subtilisin E of Bacillus subtilis (PDB ID: 3WHI), and different methods checked its quality. For the first time, this study reported the protein sequence for psychrotrophic Apr-BO1 and brought forth its novelty among other cold-active proteases.

scholarly journals Directional changes in Levallois core technologies between Eastern Africa, Arabia, and the Levant during MIS 5

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
James Blinkhorn ◽  
Huw S. Groucutt ◽  
Eleanor M. L. Scerri ◽  
Michael D. Petraglia ◽  
Simon Blockley
Keyword(s):  
Homo Sapiens ◽  
Environmental Parameters ◽  
Detailed Comparison ◽  
Eastern Africa ◽  
Middle Pleistocene ◽  
Reduction Activity ◽  
Core Technology ◽  
Geographic Expansion ◽  
First Time ◽  
Mis 5

AbstractMarine Isotope Stage (MIS) 5, ~ 130 to 71 thousand years ago, was a key period for the geographic expansion of Homo sapiens, including engagement with new landscapes within Africa and dispersal into Asia. Occupation of the Levant by Homo sapiens in MIS 5 is well established, while recent research has documented complementary evidence in Arabia. Here, we undertake the first detailed comparison of Levallois core technology from eastern Africa, Arabia, and the Levant during MIS 5, including multiple sites associated with Homo sapiens fossils. We employ quantitative comparisons of individual artefacts that provides a detailed appraisal of Levallois reduction activity in MIS 5, thereby enabling assessment of intra- and inter-assemblage variability for the first time. Our results demonstrate a pattern of geographically structured variability embedded within a shared focus on centripetal Levallois reduction schemes and overlapping core morphologies. We reveal directional changes in core shaping and flake production from eastern Africa to Arabia and the Levant that are independent of differences in geographic or environmental parameters. These results are consistent with a common cultural inheritance between these regions, potentially stemming from a shared late Middle Pleistocene source in eastern Africa.

scholarly journals Isolation and Characterization of Nitrate Reducing Bacteria for Conversion of Vegetable-Derived Nitrate to ‘Natural Nitrite’

2021 ◽  
Vol 1 (1) ◽  
pp. 11-23
Author(s):  
Arjun Bhusal ◽  
Peter M. Muriana
Keyword(s):  
Nitrate Reduction ◽  
High Performance ◽  
Limit Of Detection ◽  
Reversed Phase ◽  
Test Tube ◽  
Screening Assay ◽  
Isolation And Characterization ◽  
The Us ◽  
Reducing Bacteria ◽  
Nitrate Reducing Bacteria

In the US, sodium nitrate is used as a preservative and curing agent in processed meats and is therefore a regulated ingredient. Nitrate reducing bacteria (NRB) can convert vegetable nitrate into nitrite allowing green/clean label status in the US as per the USDA-FSIS definition of ‘natural nitrite’. The current ‘in-liquid’ test tube assay for detecting nitrite is not suitable for screening mixtures of bacteria nor is commercial nitrate broth suitable for growth of many Gram (+) bacteria. M17 broth was therefore used to develop M17-nitrate broth to be inclusive of Gram (+) bacteria. An ‘on-agar’ colony-screening assay was developed to detect the conversion of nitrate to nitrite on agar plates and could detect one NRB+ colony among ~300–500 colonies on a single plate. Samples that might have NRB were spread-plated on M17 agar plates, sandwiched with nitrate agar, and after incubation followed with sequential agar overlays containing the reagents used in the nitrate reduction assay; the appearance of red color zones above colonies indicated the presence of nitrite. NRB derived from various samples were confirmed for nitrate conversion and both nitrate and nitrite were quantified by C8 reversed-phase (RP) ion-pairing high performance liquid chromatography (HPLC) analysis (1 ppm limit of detection). Staphylococcus carnosus, a strain commonly used for nitrate reduction, was able to convert 1100 ppm M17-nitrate broth to 917 ppm nitrite. Staphylococcus caprae and Panteoa agglomerans, NRB isolated using the M17-nitrate agar assay, were also able to ferment the same broth to 916 ppm and 867 ppm nitrite, respectively. This is the first report of an on-agar colony screening assay for the detection and isolation of nitrite reducing bacteria allowing NRB to be readily isolated. This may allow for the identification of new bacteria that may have a more efficient process to generate nitrite, and possibly concomitant with production of additional natural antimicrobials, as vegetable nitrite becomes more widely used to prevent spore germination.

A water-soluble hybrid[4]arene: synthesis, host–guest complexation and application in the construction of a supra-amphiphile

2016 ◽  
Vol 40 (5) ◽  
pp. 4756-4760 ◽  
Author(s):  
Bin Hua ◽  
Li Shao ◽  
Jiong Zhou ◽  
Guocan Yu
Keyword(s):  
Water Soluble ◽  
Ph Responsive ◽  
New Host ◽  
Recognition Motif ◽  
First Time

A water-soluble hybrid[4]arene was synthesized for the first time and its pH-responsive host–guest complexation with paraquat in water was investigated. This new host–guest recognition motif was further applied in the construction of a supra-amphiphile.

scholarly journals Characterization of a Novel Fructosyltransferase from Lactobacillus reuteri That Synthesizes High-Molecular-Weight Inulin and Inulin Oligosaccharides

2002 ◽  
Vol 68 (9) ◽  
pp. 4390-4398 ◽  
Author(s):  
S. A. F. T. van Hijum ◽  
G. H. van Geel-Schutten ◽  
H. Rahaoui ◽  
M. J. E. C. van der Maarel ◽  
L. Dijkhuizen
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Lactobacillus Reuteri ◽  
Bacterial Strains ◽  
Potential Health ◽  
Isolation And Characterization ◽  
A Cell ◽  
Encoding Gene ◽  
First Time

ABSTRACT Fructosyltransferase (FTF) enzymes produce fructose polymers (fructans) from sucrose. Here, we report the isolation and characterization of an FTF-encoding gene from Lactobacillus reuteri strain 121. A C-terminally truncated version of the ftf gene was successfully expressed in Escherichia coli. When incubated with sucrose, the purified recombinant FTF enzyme produced large amounts of fructo-oligosaccharides (FOS) with β-(2→1)-linked fructosyl units, plus a high-molecular-weight fructan polymer (>107) with β-(2→1) linkages (an inulin). FOS, but not inulin, was found in supernatants of L. reuteri strain 121 cultures grown on medium containing sucrose. Bacterial inulin production has been reported for only Streptococcus mutans strains. FOS production has been reported for a few bacterial strains. This paper reports the first-time isolation and molecular characterization of (i) a Lactobacillus ftf gene, (ii) an inulosucrase associated with a generally regarded as safe bacterium, (iii) an FTF enzyme synthesizing both a high molecular weight inulin and FOS, and (iv) an FTF protein containing a cell wall-anchoring LPXTG motif. The biological relevance and potential health benefits of an inulosucrase associated with an L. reuteri strain remain to be established.

scholarly journals Live-Cell Imaging of Caspase Activation for High-Content Screening

2009 ◽  
Vol 14 (8) ◽  
pp. 956-969 ◽  
Author(s):  
Christophe Antczak ◽  
Toshimitsu Takagi ◽  
Christina N. Ramirez ◽  
Constantin Radu ◽  
Hakim Djaballah
Keyword(s):  
Cell Death ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Exposure Period ◽  
Live Cells ◽  
Assay Method ◽  
Caspase Activation ◽  
Fluorogenic Substrate ◽  
Automated Imaging ◽  
First Time

Caspases are central to the execution of programmed cell death, and their activation constitutes the biochemical hallmark of apoptosis. In this article, the authors report the successful adaptation of a high-content assay method using the DEVDNucView488™ fluorogenic substrate, and for the first time, they show caspase activation in live cells induced by either drugs or siRNA. The fluorogenic substrate was found to be nontoxic over an exposure period of several days, during which the authors demonstrate automated imaging and quantification of caspase activation of the same cell population as a function of time. Overexpression of the antiapoptotic protein Bcl-XL, alone or in combination with the inhibitor Z-VAD-FMK, attenuated caspase activation in HeLa cells exposed to doxorubicin, etoposide, or cell death siRNA. This method was further validated against 2 well-characterized NSCLC cell lines reported to be sensitive (H3255) or refractory (H2030) to erlotinib, where the authors show a differential time-dependent activation was observed for H3255 and no significant changes in H2030, consistent with their respective chemosensitivity profile. In summary, the results demonstrate the feasibility of using this newly adapted and validated high-content assay to screen chemical or RNAi libraries for the identification of previously uncovered enhancers and suppressors of the apoptotic machinery in live cells. ( Journal of Biomolecular Screening 2009:956-969)

Sign in / Sign up

Export Citation Format

Share Document