scholarly journals Use of Nonspecific, Glutamic Acid-Free, Media and High Glycerol or High Amylase as Inducing Parameters for Screening Bacillus Isolates Having High Yield of Polyglutamic Acid

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Nandita N. Baxi
Keyword(s):  
Glutamic Acid ◽  
Synthetic Medium ◽  
Maximum Yield ◽  
Proteinase K ◽  
High Yield ◽  
Polyglutamic Acid ◽  
Production Medium ◽  
Coomassie Blue ◽  
Sds Page ◽  
Free Media

Out of fifty-five Bacillus isolates obtained from ten different regional locations and sources, seven showed the ability to consistently produce specific extracellular polymeric substance (EPS) on rich as well as synthetic but nonspecific media which did not contain glutamic acid. The isolates were identified as either Bacillus licheniformis or Bacillus subtilis. The EPS from all isolates was resistant to alpha protease, proteinase K, and was thus of high molecular weight. Further it was detected after SDS-PAGE by methylene blue but not by coomassie blue R staining as in case of proteins with high proportion of acidic amino acids. Cell-free EPS, after acid hydrolysis, showed absence of carbohydrates and presence of only glutamic acid. Thus the native the EPS from all seven isolates was confirmed to be gamma polyglutamic acid (PGA) and not exopolysaccharide. The Bacillus isolate T which produced maximum polymer on all media tested had higher amylase: protease activity as compared to other strains. If inoculum was developed in rich medium as compared to synthetic medium, the PGA produced increased by twofold in the subsequent synthetic production medium. Similarly, use of inoculum consisting of young and vegetative cells also increased the PGA production by twofold though amount of inoculum did not affect yield of PGA. Though PGA was produced in even in the absence of glutamic acid supplementation in the production medium by all isolates, the yield of PGA increased by fourfold in the presence glutamic acid and the maximum yield was 30 g/l for isolate K. The supplementation of glutamine instead of glutamic acid into the medium caused an increase in the viscosity of the non-Newtonian solution of PGA.

Growth of Aphanomyces cochlioides in synthetic media as affected by carbon, nitrogen, methionine, and trace elements

1973 ◽  
Vol 51 (12) ◽  
pp. 2495-2503 ◽  
Author(s):  
Leonard J. Herr
Keyword(s):  
Trace Elements ◽  
Glutamic Acid ◽  
Synthetic Medium ◽  
Maximum Yield ◽  
Major Elements ◽  
Aphanomyces Cochlioides ◽  
N Source ◽  
Trace Element Nutrition ◽  
Methionine Concentration ◽  
Synthetic Media

Aspects of the nutrition of Aphanomyces cochlioides were investigated using a synthetic medium containing the major elements (ME) C, N, S, Ca, K, Mg, P; with or without the trace elements (TE) Fe, Zn, Cu, B, Mn, Mo. Primary C, N, and S sources were D-glucose, L-asparagine or DL-glutamic acid, and DL-methionine. In the presence or absence of individual trace elements, growth increased with Fe, decreased with Cu, and was unaffected by the remaining trace elements. Combining Fe with Cu eliminated the toxicity of Cu. C, N, and methionine concentration level effects were studied in relation to trace element nutrition and N sources in a series of factorial experiments. Trace elements strongly affected growth (maximum yield minus TE = 21.0 mg; maximum yield plus TE = 139.7 mg). As a source of C, glucose supported greater growth (83 mg) than sucrose (18 mg), while growth without added C was poor (7 mg). L-Asparagine was superior to DL-glutamic acid as a source of N at high C levels and the effect of methionine concentrations on growth differed with the N source. D-Asparagine was not utilized as a source of N. Growth in the synthetic medium approached that obtained in a complex medium.

scholarly journals Enrichment of Exosome-Like Extracellular Vesicles from Plasma Suitable for Clinical Vesicular miRNA Biomarker Research

2019 ◽  
Vol 8 (11) ◽  
pp. 1995 ◽  
Author(s):  
Moon ◽  
Shin ◽  
Kim ◽  
Lee ◽  
Mankhong ◽  
...  
Keyword(s):  
Human Plasma ◽  
Extracellular Vesicles ◽  
Proteinase K ◽  
High Yield ◽  
Clinical Samples ◽  
Diagnostic Biomarkers ◽  
Micro Rnas ◽  
Biomarker Research ◽  
Protein Rich ◽  
Yield And Purity

Exosome-like extracellular vesicles (ELVs) contain biomolecules that have potential as diagnostic biomarkers, such as proteins, micro-RNAs (miRNAs), and lipids. However, it is difficult to enrich ELVs consistently with high yield and purity from clinical samples, which hampers the development of ELV biomarkers. This is particularly true for miRNAs in protein-rich plasma. Hence, we modified ELV isolation protocols of three commercially available polymer-precipitation-based kits using proteinase K (PK) treatment to quantify ELV-associated miRNAs in human plasma. We compared the yield, purity, and characteristics of enriched plasma ELVs, and measured the relative quantity of three selected miRNAs (miR-30c, miR-126, and miR-192) in ELVs using six human plasma samples. Compared with the original protocols, we demonstrated that ELVs can be isolated with PK treatment with high purity (i.e., lack of non-exosomal proteins and homogeneous size of vesicles) and yield (i.e., abundancy of exosomal markers), which were dependent on kits. Using the kit with the highest purity and yield with PK treatment, we successfully quantified ELV miRNAs (levels of 45%–65% in total plasma) with acceptable variability. Collectively, ELV enrichment using the modified easy-to-use method appears suitable for the analysis of miRNAs, although its clinical applicability needs to be confirmed in larger clinical studies.

scholarly journals Promoting Cell Growth And Poly-Y-Glutamic Acid Production By Boosting The Synthesis of Alanine And D-Alanyl-D-Alanine In Bacillus Licheniformis

2022 ◽  
Author(s):  
Zheng Zhang ◽  
Penghui He ◽  
Shiying Hu ◽  
Yanqing Yu ◽  
Xiaoting Wang ◽  
...  
Keyword(s):  
Cell Growth ◽  
Microbial Biomass ◽  
Glutamic Acid ◽  
Bacillus Licheniformis ◽  
Alanine Racemase ◽  
High Yield ◽  
Effective Strategy ◽  
Genes Encoding ◽  
Alanine Synthesis ◽  
Glutamic Acid Production

Abstract Objective: The production of some bio-chemicals affected by the cell growth. This study aimed at promoting the cell growth by overexpressing the synthesis of peptidoglycans tetrapeptide tail components to improve poly-γ-glutamic acid (γ-PGA) production. Results: L-alanine, D-alanine and D-alanyl-D-alanine are primary precursors for the synthesis of peptidoglycans. The addition of L-alanine and D-alanine significantly increased both the cell growth and production of γ-PGA. Then, several genes encoding key enzymes for L/D-alanine and D-alanyl-D-alanine biosynthesis were overexpressed respectively, including ald (encoding alanine dehydrogenase), dal (encoding alanine racemase) and ddl (encoding D-alanine ligase). The results showed that the overexpression of genes ald , dal and ddl increased the production of γ-PGA by 19.72%, 15.91% and 60.90%, and increased the microbial biomass by 15.58%, 18.34% and 49.85%, respectively. Moreover, we demonstrated that the overexpression of genes ald , dal and ddl increased γ-PGA production mainly by enhancing cell growth rather than providing more precursors. Conclusions: This work illustrated the importance of the L/D-alanine and D-alanyl-D-alanine synthesis to the cell growth and the high yield of γ-PGA, and provided an effective strategy for producing γ-PGA .

scholarly journals Antigen Similarity In Hydatid Cyst Wall And Human Bone Tumours: A Short Report

2021 ◽  
Author(s):  
Maryam Hajizadeh ◽  
Fariba Amni ◽  
Maryam Sahlolbei ◽  
Masoumeh Tavakoli-yaraki ◽  
Amirreza Javadi Mamaghani ◽  
...  
Keyword(s):  
Hydatid Cyst ◽  
Cyst Wall ◽  
Bone Tumours ◽  
Short Report ◽  
Hydatid Cysts ◽  
Serum Samples ◽  
Coomassie Blue ◽  
Sds Page ◽  
Specific Antigens ◽  
Laminated Layer

Abstract Background: less studies have been done on bone cancers which are complex despite lower incidance. Hydatidosis is a parasitic disease that may influence host immunity by mimicking cancer cells antigens. So, this study aimed to elavuate the similarity of the immunogenic antigens between hydatid cyst and different bone cancers.Method: Cyst wall of hydatid cysts were collected and their antigens were separated with SDS-PAGE gel electrophoresis (SDS-PAGE). Serum samples obtained from patients with bone cancers and the anitigenicity of isolated anitgens were evaluated inwith E. granulosus( Larval form )infection and healthy individuals using western-blot approaches. Results: The crude extract of the laminated layer showed two specific antigens, 53 KDa and 70 KDa, after stainging the membrane with Coomassie blue. Both antigens reacted with the serum of different bone cancers but only the 53 KDa band reacted with all sera.Conclusion: It seems people with bone tumours may have extra antibodies in their serum comparing to healthy and hydatidosis which may be an autoantibodies; and the presence of this antibody against 70 KDa band protein in sera of patients with various types of bone cancers, may be helpful in diagnostic test or designing of preventive approaches in future.

scholarly journals The role of positively charge poly-L-lysine in the formation of high yield gold nanoplates on the surface for plasmonic sensing application

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259730
Author(s):  
Marlia Morsin ◽  
Suratun Nafisah ◽  
Rahmat Sanudin ◽  
Nur Liyana Razali ◽  
Farhanahani Mahmud ◽  
...  
Keyword(s):  
Substrate Surface ◽  
Surrounding Medium ◽  
Resonance Effect ◽  
Maximum Yield ◽  
Single Layer ◽  
Localized Surface Plasmon ◽  
High Yield ◽  
Synthesis Process ◽  
Wavelength Shift ◽  
Plasmonic Sensor

An anisotropic structure, gold (Au) nanoplates was synthesized using a two-step wet chemical seed mediated growth method (SMGM) directly on the substrate surface. Prior to the synthesis process, poly-l-lysine (PLL) as a cation polymer was used to enhance the yield of grown Au nanoplates. The electrostatic interaction of positive charged by PLL with negative charges from citrate-capped gold nanoseeds contributes to the yield increment. The percentage of PLL was varied from 0% to 10% to study the morphology of Au nanoplates in term of shape, size and surface density. 5% PLL with single layer treatment produce a variety of plate shapes such as hexagonal, flat rod and triangular obtained over the whole substrate surface with the estimated maximum yield up to ca. 48%. The high yield of Au nanoplates exhibit dual plasmonic peaks response that are associated with transverse and longitudinal localized surface plasmon resonance (TSPR and LSPR). Then, the PLL treatment process was repeated twice resulting the increment of Au nanoplates products to ca. 60%. The thin film Au nanoplates was further used as sensing materials in plasmonic sensor for detection of boric acid. The anisotropic Au nanoplates have four sensing parameters being monitored when the medium changes, which are peak position (wavelength shift), intensity of TSPR and LSPR, and the changes on sensing responses. The sensor responses are based on the interaction of light with dielectric properties from surrounding medium. The resonance effect produces by a collection of electron vibration on the Au nanoparticles surface after hit by light are captured as the responses. As a conclusion, it was found that the PLL treatment is capable to promote high yield of Au nanoplates. Moreover, the high yield of the Au nanoplates is an indication as excellent candidate for sensing material in plasmonic sensor.

Equine synovial fluid protein equalization via combinatorial peptide ligand libraries

2021 ◽  
Vol 1 (4) ◽  
pp. 18-22
Author(s):  
Pablo Fueyo ◽  
Marco Galleguillos ◽  
Cristóbal Dörner ◽  
Pedro A. Smith ◽  
Francisca Godoy ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Sodium Dodecyl ◽  
Electrophoretic Pattern ◽  
Peptide Ligand ◽  
Joint Pathology ◽  
Coomassie Blue ◽  
Standard Procedures ◽  
Sds Page ◽  
New Biomarkers ◽  
Synovial Fluid Protein

To gain further knowledge of the equine synovial fluid (SF) proteome, we propose a protocol based on the equalization of the relative concentrations of its proteins, which leads to the modification of the standard electrophoretic pattern revealing low-abundance proteins that otherwise remain undetected. Fresh SF samples were collected from ten macroscopically normal metacarpophalangeal joints of crossbred horses. The samples were processed using standard procedures as the control and via combinatorial peptide ligand libraries (CPLL) using low ionic forces (NaH2PO4 10 mM) at different pHs (4.0, 7.0, and 9.3) with 10% sodium dodecyl sulfate (SDS) and 25 mM DTT for protein resolubilization. Proteins were then separated by conventional 8% SDS-PAGE and stained with coomassie blue. After separation of the equalized proteins, there was a significant reduction in the albumin (the most abundant protein in the SF) and, at the same time, other protein bands arise that were not visible without CPLL processing. In addition, there was variation in the protein profiles at different pHs. The results suggest that protein equalization of the equine SF by CPLL could be a useful tool to better understand the articular homeostasis and/or for the detection of new biomarkers of joint pathology.

Front Cover: Specific glutamic acid residues in targeted proteins induce exaggerated retardations in Phos-tag SDS-PAGE migration

2017 ◽  
Vol 38 (8) ◽  
pp. NA-NA
Author(s):  
Eiji Kinoshita ◽  
Emiko Kinoshita-Kikuta ◽  
Kiyonobu Karata ◽  
Toshiki Kawano ◽  
Atsuhiro Nishiyama ◽  
...  
Keyword(s):  
Glutamic Acid ◽  
Front Cover ◽  
Page Migration ◽  
Sds Page

Biosynthesis of Polyglutamic Acid and its Application on Agriculture

2011 ◽  
Vol 183-185 ◽  
pp. 1219-1223 ◽  
Author(s):  
Xiang Yu ◽  
Min Wang ◽  
Qun Hui Wang ◽  
Xu Ming Wang
Keyword(s):  
Water Treatment ◽  
Glutamic Acid ◽  
Biodegradable Polymer ◽  
Water Soluble ◽  
Future Trends ◽  
Polyglutamic Acid ◽  
Separation And Purification ◽  
Potential Applications ◽  
Purification Methods ◽  
Fermentation Parameters

Poly γ-glutamic acid (γ-PGA) is a kind of water soluble and biodegradable polymer, which is nontoxic toward humans and the environment. Therefore, the potential applications of γ-PGA and its derivatives have been of interest in a broad range such as food, cosmetics, medicine, water treatment and agriculture. This paper summarizes γ-PGA synthesis pathways in microorganisms, producing strains, fermentation parameters, as well as separation and purification methods. In addition, the prospect of γ-PGA application on agriculture was introduced, pointing out the problems and the future trends in research and application of γ-PGA.

A MINIMAL SYNTHETIC MEDIUM SUPPORTING GROWTH OF A MICROCOCCUS SPECIES

1960 ◽  
Vol 6 (3) ◽  
pp. 251-256 ◽  
Author(s):  
I. J. McDonald
Keyword(s):  
Glutamic Acid ◽  
Aspartic Acid ◽  
Synthetic Medium ◽  
Ammonium Salts ◽  
Nutritional Requirements ◽  
Inorganic Sulphur ◽  
Relationship Of ◽  
The Relationship

Nutritional requirements of a Micrococcus species (M. freudenreichii, A.T.C.C. No. 407) were studied. The organism required glutamic acid, thiamine, biotin, magnesium, iron, and potassium for growth. Cells from such a synthetic medium were shown to contain methionine indicating that inorganic sulphur was used. Glutamic acid could not be replaced with glutamine (unheated), aspartic acid, asparagine, nor ammonium salts. The relationship of nutritional requirements of micrococci and staphylococci to classification is discussed.

Cerebrospinal fluid levels of alpha-synuclein measured using a poly-glutamic acid-modified gold nanoparticle-doped disposable neuro-biosensor system

The Analyst ◽  
2019 ◽  
Vol 144 (2) ◽  
pp. 611-621 ◽  
Author(s):  
Münteha Nur Sonuç Karaboğa ◽  
Mustafa Kemal Sezgintürk
Keyword(s):  
Parkinson’S Disease ◽  
Cerebrospinal Fluid ◽  
Glutamic Acid ◽  
Gold Nanoparticle ◽  
Alpha Synuclein ◽  
Polyglutamic Acid ◽  
Cerebrospinal Fluid Levels ◽  
System P ◽  
Biosensor System ◽  
Fluid Levels

A gold nanoparticle and polyglutamic acid-modified ITO-based biosensor system to detect alpha-synuclein, an important biomarker of Parkinson's disease.

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