scholarly journals Optimization of Cellulase Production from Bacteria Isolated from Soil

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Sonia Sethi ◽  
Aparna Datta ◽  
B. Lal Gupta ◽  
Saksham Gupta
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Source ◽  
Pseudomonas Fluorescens ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Fermentation Medium ◽  
Cellulase Production ◽  
Culture Conditions ◽  
Optimum Conditions ◽  
E Coli

Cellulase-producing bacteria were isolated from soil and identified as Pseudomonas fluorescens, Bacillus subtilIs, E. coli, and Serratia marcescens. Optimization of the fermentation medium for maximum cellulase production was carried out. The culture conditions like pH, temperature, carbon sources, and nitrogen sources were optimized. The optimum conditions found for cellulase production were 40°C at pH 10 with glucose as carbon source and ammonium sulphate as nitrogen source, and coconut cake stimulates the production of cellulase. Among bacteria, Pseudomonas fluorescens is the best cellulase producer among the four followed by Bacillus subtilis, E. coli, and Serratia marscens.

scholarly journals Protease Production from UV Mutated Bacillus subtilis

2012 ◽  
Vol 47 (1) ◽  
pp. 69-76 ◽  
Author(s):  
MG Sher ◽  
M Nadeem ◽  
Q Syed ◽  
M Irfan ◽  
S Baig
Keyword(s):  
Bacillus Subtilis ◽  
Inorganic Nitrogen ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Fermentation Medium ◽  
Maximum Activity ◽  
Inoculum Size ◽  
Protease Production ◽  
Protease Enzyme ◽  
Uv Mutation

UV mutation of the strain has significant contributation to enhance the yield of protease enzyme from Bacillus subtilis bacteria under the  cultivation conditions in submerged fermentation. The fermentation medium used for the production of protease composed of carbon  sources 1%, organic 1% or inorganic nitrogen sources 0.5% , K2HPO4 0.2 %, CaCl2 0.04% and MgSO4 0.02 % by mutated Bacillus subtilis  G-4 under the optimum parameters which are important to induce the mutated strain to produce high units of the protease, which were  temperature 37.5°C, pH 9, inoculum size 3 % v/v, glucose 1% as carbon source and peptone 1% as nitrogen source were give the maximum  455.25 + 1.66 units of protease. The results of stability studies revealed that protease of B. subtilis G-4 was stable over a broad range  of temperature (30 to 60°C) and pH (8 to 12). However, maximum activity (155.45U/ml) was observed at temperature 50°C and pH 10.  These characteristics render its potential use in detergent industries for detergent formulation.DOI: http://dx.doi.org/10.3329/bjsir.v47i1.10725 Bangladesh J. Sci. Ind. Res. 47(1), 69-76, 2012

Cellulase Production from Filamentous Fungi for Its Application in the Hydrolysis of Wheat Straw

2015 ◽  
Vol 1763 ◽  
Author(s):  
L. Toscano-Palomar ◽  
G. Montero-Alpirez ◽  
M. Stilianova-Stoytcheva ◽  
E. Vertiz-Pelaez ◽  
y E. Romero Uscanga
Keyword(s):  
Aspergillus Niger ◽  
Wheat Straw ◽  
Filamentous Fungi ◽  
Submerged Fermentation ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Cellulase Production ◽  
Culture Conditions ◽  
Mineral Medium ◽  
Cellulolytic Enzymes

ABSTRACTExtended research has been developed in the use of wheat straw (WS) as biomass for the production of biofuels (bioethanol), including the processes of degradation of cellulose by enzymatic systems. For centuries, Cellulose has been used by man; however, its enormous potential as a renewable energy source was recognized only after the discovery of cellulose degrading enzymes (cellulases). A wide variety of microorganisms can produce cellulolytic enzymes under appropriate culture conditions and among these microorganisms are filamentous fungi of the genera Trichoderma, Aspergillus, Penicillium and Fusarium. The purpose of this study was to produce cellulase enzyme from previously isolated and characterized filamentous fungi. Cellulytic fungi belonged toAspergillus flavus, Aspergillus niger, Aspergillus oryzae, Penicillium chrysogenum, Penicillium sp.,andTrichoderma harzianum.All these strains were preserved by lyophilization and also kept in sterile media (sand and soil) at 4 °C. The production of cellulases by submerged fermentation was performed in a Mandels mineral medium. The nitrogen sources were urea and ammonium sulfate. Glucose alone was used in the pre-inoculum, and dried and ground wheat straw was used in the fermentation as carbon sources. Subcultures of spore suspensions were incubated with orbital stirring (120 rpm) at 30 °C for 48 hours and used as inoculum for submerged fermentation with wheat straw as substrate in mineral medium with an initial pH of 5. Activity cellulase was determined by the method of 3,5-dinitrosalicylic acid (DNS). The results showed that wheat straw have potential for use as a substrate in the production of cellulases.Aspergillus nigershowed the highest enzymatic activity from the cellulase produced 0.051 FPU (filter paper units) after 96 hours of fermentation.

scholarly journals 4-Aminobutyrate (GABA) transporters from the amine-polyamine-choline superfamily: substrate specificity and ligand recognition profile of the 4-aminobutyrate permease from Bacillus subtilis

1998 ◽  
Vol 333 (3) ◽  
pp. 565-571 ◽  
Author(s):  
Casey E. BRECHTEL ◽  
Steven C. KING
Keyword(s):  
Bacillus Subtilis ◽  
Culture Conditions ◽  
Ligand Recognition ◽  
Open Chain ◽  
Conformationally Constrained ◽  
E Coli ◽  
Gaba Transporters ◽  
Transport Defect ◽  
Gaba Analogues ◽  
Limited Culture

A previous study [Ferson, Wray and Fisher (1996) Mol. Microbiol. 22, 693–701] has shown that transposon-mediated disruption of a protein 47% identical to the Escherichia coli GABA (4-aminobutyrate) transporter abolishes the ability of nitrogen-limited culture conditions to induce expression of a GABA transport activity in Bacillus subtilis. Here it is demonstrated directly that the B. subtilis GABA permease (gabP) gene can complement the transport defect in the gabP-negative E. colistrain. Unexpectedly, the ligand-recognition profile of the B. subtilis GabP was found to differ substantially from that of the highly homologous E. coli GabP. Unlike the E. coli GabP, the B. subtilis GabP: (i) exhibits approx. equal preference for the 3-carbon (β-alanine, Km = 9.6 µM) and the 4-carbon (GABA, Km = 37 µM) amino acids, and (ii) resists inhibition by bulky, conformationally constrained compounds (e.g. nipecotic acid, guvacine), which are active against GABA transporters from brain. The present study shows additionally that the B. subtilis GabP can translocate several open-chain GABA analogues (3-aminobutyrate, 3-aminopropanoate, cis-4-aminobutenoate) across the membrane via counterflow against [3H]GABA. Thus, consistent with the idea that the ligand-recognition domain of the B. subtilis GabP is less spacious than that of the close homologue from E. coli, the former exhibits more stringent requirements than the latter for substrate recognition and translocation. These distinct functional characteristics of the E. coli and B. subtilis GABA transporters provide a basis by which to identify ligand-recognition domains within the amine-polyamine-choline transporter superfamily.

scholarly journals Effect of culture conditions on nitrogen-fixing activity of bacteria isolated from cassava cultivated soils of Vietnam

2021 ◽  
Vol 43 (3) ◽  
pp. 27-35
Author(s):  
Pham Viet Cuong ◽  
Nguyen Phuong Hoa
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Source ◽  
16S Rrna ◽  
Culture Conditions ◽  
Rrna Gene ◽  
Atmospheric Nitrogen ◽  
Bacillus Sp ◽  
Nitrogen Fixing ◽  
The 16S Rrna Gene ◽  
Cultivated Soils

The bacteria capable of fixing atmospheric nitrogen were isolated from cassava cultivated soils of Vietnam. The potential isolates were identified by analyzing the 16S rRNA gene and by morphological, biochemical, cultural characteristics. The selected isolates were assigned to the species Bacillus sp. DQT2 M17, Bacillus subtilis DTAN6 M17, and Bacillus megaterium DSHB I8. The effect of culture conditions on the nitrogen-fixing activity of three selected isolates were studied and the obtained results showed that the highest amount of accumulated ammonia was detected after 6 days of incubation at 35 oC, pH 7.0 with sucrose as a carbon source. The selected strains could be exploited as inoculants for microbial fertilizer production.

scholarly journals Wild type Bacillus subtilis treated with ethyl methyl sulphonate produced catabolite insensitive mutants with improved cellulase production

2021 ◽  
Author(s):  
Oladipo Olaniyi
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Source ◽  
Enzyme Production ◽  
Cellulase Activity ◽  
Cellulase Production ◽  
Minimal Salt Medium ◽  
Salt Medium ◽  
Production Medium ◽  
Wild Type ◽  
Ethyl Methyl

Abstract The goal of this present investigation was to mutagenize Bacillus subtilis with Ethyl Methyl Sulphonate (EMS), screen the mutants for cellulase production and evaluate the influence of different glucose concentrations on their cellulase production potentials. The wild type B. subtilis was treated with 20, 40, 60 and 80 µl of EMS and the mutants generated were screened for cellulase production in minimal salt medium containing carboxylmethylcellulose (CMC) as the carbon source. Quantitatively, cellulase activity and protein contents were determined by dinitrosalicylic acid and Lowry methods respectively. Seven mutants were developed from each of the EMS concentration bringing the total to twenty-eight from all the concentrations. Approximately 14 and 57% of the mutants developed from 40 and 60µl of EMS had higher cellulase activities than the wild type, while none of the mutants developed from 20 and 80 µl of EMS had better activities than the wild type. The supplementation of 0.2, 0.5, 1.0 and 1.5% glucose in enzyme production medium caused approximately 100, 14, 29 and 14% cellulase repression respectively in the mutants developed from 60µl EMS. Mutants MSSS02 and MSSS05 were considered as catabolite insensitive mutants because their cellulase production were enhanced in comparison to wild type.

scholarly journals Studies on poly-3-hydroxyoctanoate biosynthesis by a consortium of microorganisms

2016 ◽  
Vol 27 (1) ◽  
pp. 44-47 ◽  
Author(s):  
Mihaela Carmen Eremia ◽  
Irina Lupescu ◽  
Mariana Vladu ◽  
Maria Petrescu ◽  
Gabriela Savoiu ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Source ◽  
Pseudomonas Putida ◽  
Sole Carbon Source ◽  
Bacterial Biomass ◽  
Fermentation Medium ◽  
Energy Reserve ◽  
Bacterial Strains ◽  
Sodium Octanoate ◽  
Intracellular Energy

Abstract Polyhydroxyalcanoates (PHAs) are specifically produced by a wide variety of bacteria, as an intracellular energy reserve in the form of homo- and copolymers of [R]-β-hydroxyalkanoic acids, depending on the C source used for microorganism growth, when the cells are grown under stressing conditions. In this paper we present microbiological accumulation of poly-3-hydroxyoctanoate (PHO) by using a consortium of bacterial strains, Pseudomonas putida and Bacillus subtilis, in a rate of 3:1, grown on a fermentation medium based on sodium octanoate as the sole carbon source. The experiments performed in the above mentioned conditions led to the following results: from 18.70 g sodium octanoate (7.72 g/L in the fermentation medium) used up during the bioprocess, 3.93-3.96 g/L dry bacterial biomass and 1.834 - 1.884 g/L PHA, containing 85.83 - 86.8% PHO, were obtained.

scholarly journals Putative metabolic pathway for the bioproduction of bikaverin and intermediates thereof in the wild Fusarium oxysporum LCP531 strain

AMB Express ◽  
2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Juliana Lebeau ◽  
Thomas Petit ◽  
Laurent Dufossé ◽  
Yanis Caro
Keyword(s):  
Metabolic Pathway ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Pharmaceutical Products ◽  
Culture Conditions ◽  
Fusarium Fujikuroi ◽  
Submerged Cultures ◽  
In The Wild ◽  
Considerable Work ◽  
Pharmaceutical Uses

AbstractFungal naphthoquinones, like red bikaverin, are of interest due to their growing applications in designing pharmaceutical products. Though considerable work has been done on the elucidation of bikaverin biosynthesis pathway in Fusarium fujikuroi, very few reports are available regarding its bioproduction in F. oxysporum. We are hereby proposing a putative metabolic pathway for bikaverin bioproduction in a wild F. oxysporum strain by cross-linking the pigment profiles we obtained under two different fermentation conditions with literature. Naphthoquinone pigments were extracted with a pressurized liquid extraction method, and characterized by HPLC–DAD and UHPLC-HRMS. The results led to the conclusions that the F. oxysporum LCP531 strain was able to produce bikaverin and its various intermediates, e.g., pre-bikaverin, oxo-pre-bikaverin, dinor-bikaverin, me-oxo-pre-bikaverin, and nor-bikaverin, in submerged cultures in various proportions. To our knowledge, this is the first report of the isolation of these five bikaverin intermediates from F. oxysporum cultures, providing us with steady clues for confirming a bikaverin metabolic pathway as well as some of its regulatory patterns in the F. oxysporum LCP531 strain, based on the previously reported model in F. fujikuroi. Interestingly, norbikaverin accumulated along with bikaverin in mycelial cells when the strain grew on simple carbon and nitrogen sources and additional cofactors. Along bikaverin production, we were able to describe the excretion of the toxin beauvericin as main extrolite exclusively in liquid medium containing complex nitrogen and carbon sources, as well as the isolation of ergosterol derivate in mycelial extracts, which have potential for pharmaceutical uses. Therefore, culture conditions were also concluded to trigger some specific biosynthetic route favoring various metabolites of interest. Such observation is of great significance for selective production of pigments and/or prevention of occurrence of others (aka mycotoxins).

scholarly journals β-Fructofuranosidase andβ–D-Fructosyltransferase from NewAspergillus carbonariusPC-4 Strain Isolated from Canned Peach Syrup: Effect of Carbon and Nitrogen Sources on Enzyme Production

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Gustavo Carvalho do Nascimento ◽  
Ryhára Dias Batista ◽  
Claudia Cristina Auler do Amaral Santos ◽  
Ezequiel Marcelino da Silva ◽  
Fabrício Coutinho de Paula ◽  
...  
Keyword(s):  
Yeast Extract ◽  
Soybean Protein ◽  
Low Cost ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Culture Conditions ◽  
Carbon And Nitrogen ◽  
Production Values ◽  
Pure Carbon ◽  
Fermentation Parameters

β-fructofuranosidase (invertase) andβ-D-fructosyltransferase (FTase) are enzymes used in industrial processes to hydrolyze sucrose aiming to produce inverted sugar syrup or fructooligosaccharides. In this work, a blackAspergillussp. PC-4 was selected among six filamentous fungi isolated from canned peach syrup which were initially screened for invertase production. Cultivations with pure carbon sources showed that invertase and FTase were produced from glucose and sucrose, but high levels were also obtained from raffinose and inulin. Pineapple crown was the best complex carbon source for invertase (6.71 U/mL after 3 days of cultivation) and FTase production (14.60 U/mL after 5 days of cultivation). Yeast extract and ammonium chloride nitrogen sources provided higher production of invertase (6.80 U/mL and 6.30 U/mL, respectively), whereas ammonium nitrate and soybean protein were the best nitrogen sources for FTase production (24.00 U/mL and 24.90 U/mL, respectively). Fermentation parameters for invertase using yeast extract wereYP/S= 536.85 U/g andPP= 1.49 U/g/h. FTase production showed values ofYP/S= 2,627.93 U/g andPP= 4.4 U/h using soybean protein. The screening for best culture conditions showed an increase of invertase production values by 5.10-fold after 96 h cultivation compared to initial experiments (fungi bioprospection), while FTase production increased by 14.60-fold (44.40 U/mL) after 168 h cultivation.A. carbonariusPC-4 is a new promising strain for invertase and FTase production from low cost carbon sources, whose synthesized enzymes are suitable for the production of inverted sugar, fructose syrups, and fructooligosaccharides.

scholarly journals Controlling Instability in gacS-gacARegulatory Genes during Inoculant Production of Pseudomonas fluorescens Biocontrol Strains

2000 ◽  
Vol 66 (8) ◽  
pp. 3142-3150 ◽  
Author(s):  
Brion K. Duffy ◽  
Geneviève Défago
Keyword(s):  
Pseudomonas Fluorescens ◽  
Response Regulator ◽  
Spontaneous Mutation ◽  
Scale Up ◽  
Screening Method ◽  
Carbon Sources ◽  
Transcriptional Response ◽  
Culture Conditions ◽  
Ammonium Molybdate ◽  
Spontaneous Mutants

ABSTRACT Secondary metabolism in fluorescent pseudomonads is globally regulated by gacS, which encodes a membrane-bound sensor kinase, and gacA, which encodes a transcriptional response regulator. Spontaneous mutation in either gene blocked biosynthesis of the antimicrobial compounds hydrogen cyanide, 2,4-diacetylphloroglucinol, pyoluteorin, and pyrrolnitrin by the model biocontrol strain Pseudomonas fluorescens CHA0. Spontaneous mutants also had altered abilities to utilize several carbon sources and to increase medium pH compared with the wild type, suggesting thatgacS and gacA influence primary as well as secondary bacterial metabolism. Inoculant efficacy for biocontrol was significantly reduced by contamination with regulatory mutants which accumulated during inoculum production. Spontaneous mutants accumulated in all 192 separate liquid cultures examined, typically at a frequency of 1% or higher after 12 days. During scale-up in a simulated industrial fermentation process, mutants increased exponentially and accounted for 7, 23, and 61% of the total viable cells after transfer to 20-, 100-, and 500-ml preparations, respectively. GacS−and GacA− mutants had identical phenotypes and occurred at the same frequency, indicating that the selective pressures for the two mutants were similar. We developed a simple screening method for monitoring inoculant quality based on the distinctive appearance of mutant colonies (i.e., orange color, enlarged diameter, hyperfluorescence). Mutant competitiveness was favored in a nutrient-rich medium with a high electrolyte concentration (nutrient broth containing yeast extract). We were able to control mutant accumulation and to clean up contaminated cultures by using certain mineral amendments (i.e., zinc, copper, cobalt, manganese, and ammonium molybdate) or by diluting media 1/10. Spontaneous mutants and genetic constructs had the same response to culture conditions. Zinc and medium dilution were also effective for improving the genetic stability of other P. fluorescens biocontrol strains obtained from Ghana and Italy.

scholarly journals Comparison of methods for the identification and sub-typing of O157 and non-O157 Escherichia coli serotypes and their integration into a polyphasic taxonomy approach

2016 ◽  
Vol 55 (2) ◽  
pp. 81-90 ◽  
Author(s):  
M.A. Prieto-Calvo ◽  
M.K. Omer ◽  
O. Alvseike ◽  
M. López ◽  
A. Alvarez-Ordóñez ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Carbon Sources ◽  
23S Rrna ◽  
Local Alignment ◽  
Taxonomic Structure ◽  
Sequencing Data ◽  
E Coli ◽  
Additional Information ◽  
Ft Ir

AbstractPhenotypic, chemotaxonomic and genotypic data from 12 strains ofEscherichia coli werecollected, including carbon source utilisation profiles, ribotypes, sequencing data of the 16S–23S rRNA internal transcribed region (ITS) and Fourier transform-infrared (FT-IR) spectroscopic profiles. The objectives were to compare several identification systems forE. coliand to develop and test a polyphasic taxonomic approach using the four methodologies combined for the sub-typing of O157 and non-O157E. coli. The nucleotide sequences of the 16S–23S rRNA ITS regions were amplified by polymerase chain reaction (PCR), sequenced and compared with reference data available at the GenBank database using the Basic Local Alignment Search Tool (BLAST) . Additional information comprising the utilisation of carbon sources, riboprint profiles and FT-IR spectra was also collected. The capacity of the methods for the identification and typing ofE. colito species and subspecies levels was evaluated. Data were transformed and integrated to present polyphasic hierarchical clusters and relationships. The study reports the use of an integrated scheme comprising phenotypic, chemotaxonomic and genotypic information (carbon source profile, sequencing of the 16S–23S rRNA ITS, ribotyping and FT-IR spectroscopy) for a more precise characterisation and identification ofE. coli. The results showed that identification ofE. colistrains by each individual method was limited mainly by the extension and quality of reference databases. On the contrary, the polyphasic approach, whereby heterogeneous taxonomic data were combined and weighted, improved the identification results, gave more consistency to the final clustering and provided additional information on the taxonomic structure and phenotypic behaviour of strains, as shown by the close clustering of strains with similar stress resistance patterns.

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