scholarly journals A Rapid Method for Performing a Multivariate Optimization of Phage Production Using the RCCD Approach

Pathogens ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1100
Author(s):  
Jessica Silva ◽  
Roberto Dias ◽  
José Ivo Junior ◽  
Maraísa Marcelino ◽  
Mirelly Silva ◽  
...  
Keyword(s):  
Protein Delivery ◽  
Large Scale ◽  
Minimal Medium ◽  
Culture Media ◽  
Scale Up ◽  
Carbon Sources ◽  
Luria Bertani ◽  
Luria Bertani Broth ◽  
Phage Production ◽  
Set Up

Bacteriophages can be used in various applications, from the classical approach as substitutes for antibiotics (phage therapy) to new biotechnological uses, i.e., as a protein delivery vehicle, a diagnostic tool for specific strains of bacteria (phage typing), or environmental bioremediation. The demand for bacteriophage production increases daily, and studies that improve these production processes are necessary. This study evaluated the production of a T4-like bacteriophage vB_EcoM-UFV09 (an E. coli-infecting phage with high potential for reducing environmental biofilms) in seven types of culture media (Luria–Bertani broth and the M9 minimal medium with six different carbon sources) employing four cultivation variables (temperature, incubation time, agitation, and multiplicity of infection). For this purpose, the rotatable central composite design (RCCD) methodology was used, combining and comparing all parameters to determine the ideal conditions for starting to scale up the production process. We used the RCCD to set up the experimental design by combining the cultivation parameters in a specific and systematic way. Despite the high number of conditions evaluated, the results showed that when specific conditions were utilized, viral production was effective even when using a minimal medium, such as M9/glucose, which is less expensive and can significantly reduce costs during large-scale phage production.

Evaluation of the growth and sporulation of different entomopathogenic fungi in different liquid and solid media at varied concentrations

2017 ◽  
Vol 6 (8) ◽  
pp. 5459
Author(s):  
Chandra Teja K. ◽  
Rahman S. J.
Keyword(s):  
Entomopathogenic Fungi ◽  
Large Scale ◽  
Insect Pests ◽  
Culture Media ◽  
Virulent Strain ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Maximum Growth ◽  
Nutritional Requirements ◽  
Virulent Strains

Entomopathogenic fungi like Beauveria bassiana, Metarhizium anisopliae and Lecanicillium lecanii are used in biological control of agricultural insect pests. Their specific mode of action makes them an effective alternative to the chemical Insecticides. Virulent strains of Entomopathogenic fungi are effectively formulated and used as bio-insecticides world-wide. Amenable and economical multiplication of a virulent strain in a large scale is important for them to be useful in the field. Culture media plays a major role in the large-scale multiplication of virulent strains of Entomopathogens. Different substrates and media components are being used for this purpose. Yet, each strain differs in its nutritional requirements for the maximum growth and hence it is necessary to standardize the right components and their optimum concentrations in the culture media for a given strain of Entomopathogen. In the current study, three different nitrogen sources and two different carbon sources were tried to standardize the mass multiplication media for seven test isolates of Entomopathogenic fungi. A study was also conducted to determine the ideal grain media for the optimum conidial yields of the test isolates. Yeast extract was found to be the best Nitrogen source for the isolates. The isolates tested, differed in their nutritional requirements and showed variation in the best nitrogen and carbon sources necessary for their growth. Variation was also found in the optimum concentration of both the ingredients for the growth and sporulation of the isolates. In the solid-state fermentation study, rice was found to be the best grain for the growth of most of the fungi followed by barley. The significance of such a study in the development of an effective Myco-insecticide is vital and can be successfully employed in agriculture is discussed.

scholarly journals Impact of Different Carbon Sources on the in vitro Growth and Viability of Escherichia coli (SUBE01) and Salmonella spp. (SUBS01) Cells

2016 ◽  
pp. 39-44
Author(s):  
Ifra Tun Nur ◽  
Jannatun Tahera ◽  
Md Sakil Munna ◽  
M Majibur Rahman ◽  
Rashed Noor
Keyword(s):  
Escherichia Coli ◽  
Bacterial Growth ◽  
Culture Media ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Growth Dynamics ◽  
Luria Bertani ◽  
Tween 20 ◽  
Salmonella Spp

With a previous observation of Escherichia coli growth cessation along with temperature variation within three different bacteriological culture media (nutrient agar, Luria-Bertani agar and minimal agar), current investigation further depicted on the possible growth dynamics of Escherichia coli (SUBE01) and Salmonella (SUBS01) growth and viability upon supplementation of different carbon sources (dextrose, sucrose, lactose, glycerol and tween 20) at 37°C under the aeration of 100 rpm. Viability of the tested bacterial species was assessed through the enumeration of the colony forming unit (cfu) appeared upon prescribed incubation for 12-24 hours on different agar plates consisting of the above mentioned carbon sources. Besides, to inspect the cellular phenotypic changes, morphological observations were conducted under the light microscope. Variations in bacterial growth (either growth acceleration or cessation) were further noticed through the spot tests on the agar plates. Considerable shortfalls in the culturable cells of E. coli and Salmonella spp. were noted in the minimal media separately consisting of sucrose, lactose, glycerol or tween 20 while an opposite impact of accelerated growth was noticed in the media supplied with dextrose. The data revealed a hierarchy of consequence of carbon sources as nutrient generators whereby the favourable bacterial growth and survival order of the carbon sources was estimated as dextrose > glycerol > lactose > tween 20 > sucrose.Bangladesh J Microbiol, Volume 32, Number 1-2,June-Dec 2015, pp 39-44

scholarly journals Role of the sRNA GcvB in regulation of cycA in Escherichia coli

Microbiology ◽  
2009 ◽  
Vol 155 (1) ◽  
pp. 106-114 ◽  
Author(s):  
Sarah C. Pulvermacher ◽  
Lorraine T. Stauffer ◽  
George V. Stauffer
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Minimal Medium ◽  
Luria Bertani ◽  
Cell Physiology ◽  
Rt Pcr ◽  
Luria Bertani Broth ◽  
Increased Sensitivity ◽  
Transcriptional Fusions

In Escherichia coli, the gcvB gene encodes a small non-translated RNA that regulates several genes involved in transport of amino acids and peptides (including sstT, oppA and dppA). Microarray analysis identified cycA as an additional regulatory target of GcvB. The cycA gene encodes a permease for the transport of glycine, d-alanine, d-serine and d-cycloserine. RT-PCR confirmed that GcvB and the Hfq protein negatively regulate cycA mRNA in cells grown in Luria–Bertani broth. In addition, deletion of the gcvB gene resulted in increased sensitivity to d-cycloserine, consistent with increased expression of cycA. A cycA : : lacZ translational fusion confirmed that GcvB negatively regulates cycA expression in Luria–Bertani broth and that Hfq is required for the GcvB effect. GcvB had no effect on cycA : : lacZ expression in glucose minimal medium supplemented with glycine. However, Hfq still negatively regulated the fusion in the absence of GcvB. A set of transcriptional fusions of cycA to lacZ identified a sequence in cycA necessary for regulation by GcvB. Analysis of GcvB identified a region complementary to this region of cycA mRNA. However, mutations predicted to disrupt base-pairing between cycA mRNA and GcvB did not alter expression of cycA : : lacZ. A model for GcvB function in cell physiology is discussed.

scholarly journals Evaluation of the Strain Bacillus amyloliquefaciens YP6 in Phoxim Degradation via Transcriptomic Data and Product Analysis

Molecules ◽  
2019 ◽  
Vol 24 (21) ◽  
pp. 3997 ◽  
Author(s):  
Di Meng ◽  
Liyuan Zhang ◽  
Jie Meng ◽  
Qiaopeng Tian ◽  
Lixin Zhai ◽  
...  
Keyword(s):  
Bacillus Amyloliquefaciens ◽  
Culture Media ◽  
Organophosphorus Pesticide ◽  
Luria Bertani ◽  
Inoculum Size ◽  
Product Analysis ◽  
Transcriptomic Data ◽  
Luria Bertani Broth ◽  
Phosphorus Source ◽  
First Time

Phoxim, a type of organophosphorus pesticide (OP), is widely used in both agriculture and fisheries. The persistence of phoxim has caused serious environmental pollution problems. In this study, Bacillus amyloliquefaciens YP6 (YP6), which is capable of promoting plant growth and degrading broad-spectrum OPs, was used to study phoxim degradation. Different culture media were applied to evaluate the growth and phoxim degradation of YP6. YP6 can grow rapidly and degrade phoxim efficiently in Luria–Bertani broth (LB broth) medium. Furthermore, it can also utilize phoxim as the sole phosphorus source in a mineral salt medium. Response surface methodology was performed to optimize the degradation conditions of phoxim by YP6 in LB broth medium. The optimum biodegradation conditions were 40 °C, pH 7.20, and an inoculum size of 4.17% (v/v). The phoxim metabolites, O,O-diethylthiophosphoric ester, phoxom, and α-cyanobenzylideneaminooxy phosphonic acid, were confirmed by liquid chromatography–mass spectrometry. Meanwhile, transcriptome analysis and qRT-PCR were performed to give insight into the phoxim-stress response at the transcriptome level. The hydrolase-, oxidase-, and NADPH-cytochrome P450 reductase-encoding genes were significantly upregulated for phoxim hydrolysis, sulfoxidation, and o-dealkylation. Furthermore, the phoxim biodegradation pathways by YP6 were proposed, for the first time, based on transcriptomic data and product analysis.

scholarly journals Melanogenesis Using Tyrosinate (Not Tyrosinase)

2018 ◽  
Author(s):  
Koen Vercruysse ◽  
Nahfisa Richardson
Keyword(s):  
Large Scale ◽  
Culture Media ◽  
Disodium Salt ◽  
Size Exclusion ◽  
Future Research ◽  
Vis Spectroscopy ◽  
Set Up ◽  
Mediated Oxidation

<p>We present our initial observations regarding the effect of the presence of L-tyrosinate (= L-tyrosine disodium salt) on the auto- or Fe<sup>2+</sup>/H<sub>2</sub>O<sub>2</sub>-mediated oxidation of various catecholic substances into melanin-like pigments. We observed that L-tyrosinate inhibited the Fe<sup>2+</sup>/H<sub>2</sub>O<sub>2</sub>-mediated oxidation. In contrast, L-tyrosinate promoted the auto-oxidation of ortho-diphenols like L-DOPA, dopamine, epinephrine, norepinephrine, catechol or pyrogallol, but not a meta-diphenol like resorcinol. In addition, we briefly demonstrated the melanogenic properties of cell culture media containing L-tyrosinate. The reactions were monitored using UV-Vis spectroscopy and size exclusion chromatography. For a reaction between L-tyrosinate and L-DOPA, a large scale experiment was set up allowing us to isolate, purify and characterize using FT-IR spectroscopy the melanin-like material obtained. We discuss our observations in the context of the <i>in vitro</i> and <i>in vivo</i> study of melanogenesis and provide some directions for future research efforts.<i></i></p>

Comparative Analysis of Shigella boydii 18 Foodborne Outbreak Isolate and Related Enteric Bacteria: Role of rpoS and adiA in Acid Stress Response

2005 ◽  
Vol 68 (3) ◽  
pp. 521-527 ◽  
Author(s):  
YVONNE C. CHAN ◽  
HANS P. BLASCHEK
Keyword(s):  
Minimal Medium ◽  
Enteric Bacteria ◽  
Arginine Decarboxylase ◽  
Luria Bertani ◽  
Decarboxylase Activity ◽  
E Coli ◽  
Rpos Gene ◽  
Shigella Boydii ◽  
Luria Bertani Broth ◽  
K 12

Shigella boydii CDPH (Chicago Department of Public Health) serotype 18 was implicated in an outbreak of foodborne illness in 1998. The suspected food vehicles were parsley and cilantro imported from Mexico used to prepare bean salad. Previous studies revealed that S. boydii CDPH serotype 18 can survive in bean salad, which contains organic acids and whose pH decreases over time. Acid challenge assays in acidified tryptic soy broth at pH 4.5, acidified Luria-Bertani broth at pH 4.5, and acidified M9 minimal salts medium at pH 2.5 containing amino acids, arginine, or glutamic acid were performed using S. boydii CDPH, S. boydii ATCC 35966, S. flexneri 3136, Escherichia coli O157:H7 dd8872, and E. coli O157:H7 dd642 to compare differences in acid tolerance. Differences in survival of exponential-phase cells were detected in acidified tryptic soy broth and Luria-Bertani broth at pH 4.5. In acidified minimal medium containing arginine, S. boydii strains were able to survive at pH 2.5. The arginine decarboxylase gene (adiA) present in S. boydii is involved in survival at extremely low pH. The discovery of adiA expression in S. boydii serotype 18 by use of an acidified minimal medium challenge and arginine decarboxylase biochemical assay is significant because arginine decarboxylase activity was thought to be unique to E. coli. Sequencing of the rpoS gene from the S. boydii outbreak strain indicates that it is 99% conserved compared with the E. coli K-12 rpoS gene and plays a vital role in survival under acidic conditions.

scholarly journals Komagataeibacter intermedius V-05: An Acetic Acid Bacterium Isolated from Vinegar Industry, with High Capacity for Bacterial Cellulose Production in Soybean Molasses-Based Medium

2021 ◽  
Vol 59 (4) ◽  
Author(s):  
Rodrigo José Gomes ◽  
Paula Cristina de Sousa Faria-Tischer ◽  
Cesar Augusto Tischer ◽  
Leonel Vinicius Constantino ◽  
Morsyleide de Freitas Rosa ◽  
...  
Keyword(s):  
Bacterial Cellulose ◽  
Large Scale ◽  
Culture Media ◽  
Carbon Sources ◽  
High Capacity ◽  
Cellulose Synthesis ◽  
Standard Medium ◽  
Cellulose Production ◽  
Suitable Alternative

Research background. Despite the great properties of bacterial cellulose, its manufacture is still limited due to difficulties in production at large-scale. These problems are mainly related to low production yields and high overall costs of the conventional culture media normally used. Reversing these problems makes it necessary to identify new cheap and sustainable carbon sources. Thus, this work aimed to isolate and select a high cellulose-producing Komagataeibacter strain from vinegar industry, and study their potential for bacterial cellulose synthesis in an industrial soybean co-product, known as soybean molasses, to be used as fermentation medium. Experimental approach. For one isolated strain that exhibited high level of cellulose production in the standard Hestrin-Schramm medium, the ability of this biopolymer production in a soybean molasses-based medium was determined. The produced membranes were characterized by thermogravimetric analysis, X-ray diffraction, infrared spectroscopy, water holding capacity and rehydration ratio for determination of its characteristics and properties. The selected strain was also characterized by genetic analysis for determination of its genus and specie. Results and conclusions. An isolated strain was genetically identified as Komagataeibacter intermedius V-05 and exhibited the highest cellulose production in Hestrin-Schramm medium (3.7 g/L). In addition, the production by this strain in soybean molasses-based medium was 10.0 g/L. Membranes from both substrates were similar in terms of chemical structure, crystallinity and thermal degradation. Soybean molasses proved to be a suitable alternative medium for biosynthesis of cellulose in comparison with standard medium. In addition to providing higher production yield, the membranes showed great structural characteristics, similar to those obtained from standard medium. Novelty and scientific contribution. In this research, we have isolated and identified a Komagataeibacter strain which exhibits a high capacity for cellulose production in soybean molasses medium. The isolation and selection of strains with high capacity of microbial metabolites production is important for decreasing bioprocess costs. Furthermore, as there is a necessity today to find cheaper carbon sources that provide microbial products at a lower cost, soybean molasses represents an interesting alternative medium to produce bacterial cellulose prior to its industrial application.

scholarly journals Degradation of 2,3-Diethyl-5-Methylpyrazine by a Newly Discovered Bacterium, Mycobacterium sp. Strain DM-11

2006 ◽  
Vol 72 (2) ◽  
pp. 1437-1444 ◽  
Author(s):  
Sugima Rappert ◽  
Kathrin Caroline Botsch ◽  
Stephanie Nagorny ◽  
Wittko Francke ◽  
Rudolf Müller
Keyword(s):  
Energy Source ◽  
Molecular Oxygen ◽  
Carbon Sources ◽  
Treatment Plant ◽  
Luria Bertani ◽  
Ring Cleavage ◽  
Rrna Gene ◽  
Gas Treatment ◽  
Luria Bertani Broth ◽  
Waste Gas Treatment

ABSTRACT A bacterium was isolated from the waste gas treatment plant at a fishmeal processing company on the basis of its capacity to use 2,3-diethyl-5-methylpyrazine (DM) as a sole carbon and energy source. The strain, designated strain DM-11, grew optimally at 25°C and had a doubling time of 29.2 h. The strain did not grow on complex media like tryptic soy broth, Luria-Bertani broth, or nutrient broth or on simple carbon sources like glucose, acetate, oxoglutarate, succinate, or citrate. Only on Löwenstein-Jensen medium was growth observed. The 16S rRNA gene sequence of strain DM-11 showed the highest similarity (96.2%) to Mycobacterium poriferae strain ATCC 35087T. Therefore, strain DM-11 merits recognition as a novel species within the genus Mycobacterium. DM also served as a sole nitrogen source for the growth of strain DM-11. The degradation of DM by strain DM-11 requires molecular oxygen. The first intermediate was identified as 5,6-diethyl-2-hydroxy-3-methylpyrazine (DHM). Its disappearance was accompanied by the release of ammonium into the culture medium. No other metabolite was detected. We conclude that ring fission occurred directly after the formation of DHM and ammonium was eliminated after ring cleavage. Molecular oxygen was essential for the degradation of DHM. The expression of enzymes involved in the degradation of DM and DHM was regulated. Only cells induced by DM or DHM converted these compounds. Strain DM-11 also grew on 2-ethyl-5(6)-methylpyrazine (EMP) and 2,3,5-trimethylpyrazine (TMP) as a sole carbon, nitrogen, and energy source. In addition, the strain converted many pyrazines found in the waste gases of food industries cometabolically.

scholarly journals The Small RNA GcvB Regulates sstT mRNA Expression in Escherichia coli

2008 ◽  
Vol 191 (1) ◽  
pp. 238-248 ◽  
Author(s):  
Sarah C. Pulvermacher ◽  
Lorraine T. Stauffer ◽  
George V. Stauffer
Keyword(s):  
Escherichia Coli ◽  
Minimal Medium ◽  
Small Region ◽  
Luria Bertani ◽  
Negative Regulation ◽  
Transport Systems ◽  
Mrna Levels ◽  
Glucose Minimal Medium ◽  
Reverse Transcription Pcr ◽  
Luria Bertani Broth

ABSTRACT In Escherichia coli, the gcvB gene encodes a nontranslated RNA (referred to as GcvB) that regulates OppA and DppA, two periplasmic binding proteins for the oligopeptide and dipeptide transport systems. An additional regulatory target of GcvB, sstT, was found by microarray analysis of RNA isolated from a wild-type strain and a gcvB deletion strain grown to mid-log phase in Luria-Bertani broth. The SstT protein functions to transport l-serine and l-threonine by sodium transport into the cell. Reverse transcription-PCR and translational fusions confirmed that GcvB negatively regulates sstT mRNA levels in cells grown in Luria-Bertani broth. A series of transcriptional fusions identified a region of sstT mRNA upstream of the ribosome binding site needed for negative regulation by GcvB. Analysis of the GcvB RNA identified a sequence complementary to this region of the sstT mRNA. The region of GcvB complementary to sstT mRNA is the same region of GcvB identified to regulate the dppA and oppA mRNAs. Mutations predicted to disrupt base pairing between sstT mRNA and GcvB were made in gcvB, which resulted in the identification of a small region of GcvB necessary for negative regulation of sstT-lacZ. Additionally, the RNA chaperone protein Hfq was found to be necessary for GcvB to negatively regulate sstT-lacZ in Luria-Bertani broth and glucose minimal medium supplemented with glycine. The sstT mRNA is the first target found to be regulated by GcvB in glucose minimal medium supplemented with glycine.

scholarly journals Melanogenesis Using Tyrosinate (Not Tyrosinase)

2018 ◽  
Author(s):  
Koen Vercruysse ◽  
Nahfisa Richardson
Keyword(s):  
Large Scale ◽  
Culture Media ◽  
Disodium Salt ◽  
Size Exclusion ◽  
Future Research ◽  
Vis Spectroscopy ◽  
Set Up ◽  
Mediated Oxidation

<p>We present our initial observations regarding the effect of the presence of L-tyrosinate (= L-tyrosine disodium salt) on the auto- or Fe<sup>2+</sup>/H<sub>2</sub>O<sub>2</sub>-mediated oxidation of various catecholic substances into melanin-like pigments. We observed that L-tyrosinate inhibited the Fe<sup>2+</sup>/H<sub>2</sub>O<sub>2</sub>-mediated oxidation. In contrast, L-tyrosinate promoted the auto-oxidation of ortho-diphenols like L-DOPA, dopamine, epinephrine, norepinephrine, catechol or pyrogallol, but not a meta-diphenol like resorcinol. In addition, we briefly demonstrated the melanogenic properties of cell culture media containing L-tyrosinate. The reactions were monitored using UV-Vis spectroscopy and size exclusion chromatography. For a reaction between L-tyrosinate and L-DOPA, a large scale experiment was set up allowing us to isolate, purify and characterize using FT-IR spectroscopy the melanin-like material obtained. We discuss our observations in the context of the <i>in vitro</i> and <i>in vivo</i> study of melanogenesis and provide some directions for future research efforts.<i></i></p>

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