scholarly journals Enhanced production of inulinase from Xanthomonas campestris pv. phaseoli

2010 ◽  
Author(s):  
◽  
Kameshnee Naidoo
Keyword(s):  
Xanthomonas Campestris ◽  
Large Scale ◽  
Specific Activity ◽  
Carbon Sources ◽  
Gel Filtration ◽  
Catalytic Efficiency ◽  
Scale Production ◽  
Inulin Hydrolysis ◽  
Enhanced Production ◽  
Good Potential

Xanthomonas campestris pv phaseoli produced an extracellular endoinulinase on various carbon sources. The highest inulinase production of 9.24 ± 0.03 IU ml¯¹by X. campestris pv. phaseoli was attained using an optimized medium comprising of 3% sucrose and 2.5% tryptone. Inulinase production in X. campestris pv. phaseoli was further enhanced through ethylmethanesulfonate mutagenesis. The resulting mutant, X. campestris pv. phaseoli KM 24 demonstrated enhanced inulinase production of 22.09 ± 0.03 IU ml¯¹after 24 h, which was 2.4 – fold higher than that of the wild type. Inulinase production by this mutant was scaled up in a 5 L fermenter yielding a final activity of 21.87 ± 0.03 IU ml¯¹with an inulinase/invertase (I/S) ratio of 2.6 after 18 h. Maximum volumetric (21 865 IU 1¯¹ h¯¹) and specific (119 025 IU g¯¹ h¯¹) productivities of inulinase were attained in a fermenter after 18h growth. Inulin hydrolysis by the crude inulinase and subsequent detection of mono- and oligosaccharides indicated the presence of an endoinulinase. The extracellular endoinulinase from the mutant KM 24 was purified to homogeneity by gel filtration chromatography and had a specific activity of 174.74U/mg. the optimum pH and temperature of the purified enzyme were found to be 6.0 and 50°C, respectively. The enzyme was stable up to 60°C, retaining over 60% activity for 30 min, but activity rapidly declined at temperatures above 60°C. The pure inulinase enzyme was also found to be stable between pH 6-9. The Lineweaver-Burk plots showed that the apparent Km and Vmax values of the inulinase for inulin were 1.15 mg/ml and 15µM/min, respectively. The Kcat value was found to be 0.145 min¯¹ with an enzyme catalytic efficiency of 0.126 mg¯¹.ml.min¯¹.This mutant demonstrated good potential for large scale production of inulinase and fructooligosaccharides.

Large-scale production of citrate synthase from a cloned gene

1982 ◽  
Vol 60 (12) ◽  
pp. 1143-1147 ◽  
Author(s):  
Harry W. Duckworth ◽  
Alexander W. Bell
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Citrate Synthase ◽  
Specific Activity ◽  
Tetracycline Resistance ◽  
Scale Production ◽  
Ampicillin Resistance ◽  
Colicin E1 ◽  
Large Scale Production ◽  
Cloned Gene

Starting with a colicin E1 resistance recombinant plasmid which contains gltA, the gene for citrate synthase in Escherichia coli, we have constructed an ampicillin-resistance plasmid containing the gltA region as a 2.9-kilobase-pair insert in the tetracycline-resistance region of pBR322. Escherichia coli HB101 harbouring this plasmid, when grown on rich medium containing ampicillin, contains citrate synthase as about 8% of its soluble protein. The enzyme has been purified from this rich source and is identical to the chromosomal enzyme prepared previously in every property tested, except for specific activity, which is 64 U∙mg−1 as compared with 45–50 U∙mg−1 previously obtained. The N-terminal sequences of both enzymes are reported, and they are identical up to residue 16 at least. The overall yield of pure enzyme, starting with the cells grown in 15 L of medium, is 600–800 mg.

scholarly journals Enhanced production of 99Mo in inverse kinematics heavy ion reactions

2021 ◽  
Vol 252 ◽  
pp. 08003
Author(s):  
Justin Mabiala ◽  
Marcia R.D. Rodrigues ◽  
Georgios A. Souliotis ◽  
Victor E. Iacob ◽  
Ninel Nica ◽  
...  
Keyword(s):  
Inverse Kinematics ◽  
Large Scale ◽  
Heavy Ion ◽  
Scale Production ◽  
Gas Cell ◽  
Enhanced Production ◽  
Gas Target ◽  
Large Scale Production ◽  
Cell Target ◽  
Catcher Foil

The reaction of a 100Mo beam at 12 MeV/nucleon impinging on a 4He gas-cell target was performed. The 99Mo alongside other coproduced isotopes were collected after the gas target on an aluminum catcher foil and their respective radioactivities were measured by offline γ-ray analysis. In this contribution, preliminary experimental results which are used to discuss the possibility of optimal large-scale production conditions of the produced radioisotopes are presented.

Enhanced production of falcarinol-type polyacetylenes in hairy roots of cultivated carrot (Daucus carota subsp. sativus)

2021 ◽  
Author(s):  
Frank Dunemann ◽  
Christoph Böttcher
Keyword(s):  
Hairy Root ◽  
Large Scale ◽  
Higher Plants ◽  
Leaf Tissue ◽  
Hairy Root Cultures ◽  
Scale Production ◽  
Root Cultures ◽  
Tissue Samples ◽  
Positive Effects ◽  
Enhanced Production

Abstract Polyacetylenes (PAs) are a large group of bioactive phytochemicals, which are primarily produced by higher plants of the families Apiaceae and Araliaceae. Especially aliphatic C17-polyacetylenes of the falcarinol-type such as falcarinol (FaOH) and falcarindiol (FaDOH) are known for their numerous positive effects on human health. In this study we investigate the potential of carrot hairy root cultures for production of PAs. Three individual plants of seven differently coloured carrot cultivars were used for the development of hairy root cultures by transformation of root discs with the wild-type Rhizobium rhizogenes strain 15834. A total of 51 individual hairy root (HR) lines were obtained and quantitatively analysed together with root, petiole and leaf tissue samples for FaOH and FaDOH. Among the five tissues sampled from the donor plants, root periderm samples generally exhibited the highest PA levels with FaDOH as prevailing PA and large differences between cultivars. In comparison to periderm tissue, FaOH levels were highly increased in HR lines of all cultivars. In contrast, FaDOH levels were not significantly altered. Considering the low to moderate PA concentration in root and leaf tissues of the orange cultivars there was an up to more than 10-fold increase of the FaOH concentration in HRs of these genotypes. Within this study a reproducible method for Rrhi-mediated transformation of carrot root discs was applied which provides an efficient tool to assess the function of candidate genes involved in the biosynthesis of key PAs in carrot but might be used in future also for the large-scale production of falcarinol-type PAs.

scholarly journals A New Procedure for Purification of Crotalase From Crotalus Adamanteus Venom

1977 ◽  
Author(s):  
F.S. Markland ◽  
J. Chou ◽  
Y. Shih ◽  
H. Pirkle
Keyword(s):  
Sodium Chloride ◽  
Large Scale ◽  
Specific Activity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Fold Increase ◽  
Tris Buffer ◽  
High Yield ◽  
Crude Venom ◽  
Diamondback Rattlesnake

A new procedure has been developed for large scale, rapid purification of crotalase, the thrombin-1ike enzyme from the venom of the eastern diamondback rattlesnake (Crotalus adamanteus). The three step procedure involves: (1) molecular sieve chromatography on Sephadex G-100 in 0.04 M Tris buffer containing 0.10 M sodium chloride, pH 7.1; (2) gradient elution from DEAE-cellulose with sodium acetate buffer, pH 7.0; and (3) affinity chromatography on p-aminobenzamidine Sepharose using a spacer of 6-aminohexanoic acid. Crotalase was eluted from the affinity resin by 0.05 M Tris buffer containing 0.10 M sodium chloride and 0.15 M benzamidine-hydrochloride, pH 9.0, after first washing with the Tris buffer containing 0.40 M sodium chloride. From the crude venom, pure enzyme was obtained with an overall recovery of 40-60% of clotting activity and a 90-100 fold increase in specific activity. Crotalase was shown to be pure by Polyacrylamide disk gel electrophoresis which gave one band. The molecular weight was estimated to be approximately 31,000 by gel filtration on a calibrated Sephadex G-100 column. Amino acid analysis was performed and the composition was shown to be very similar to that reported earlier (F.S. Markland and P.S. Damus, J. Biol. Chem. 246: 6460, 1971). Clotting activity of the enzyme was not inhibited by heparin, either with or without plasma, whereas, thrombin was rapidly inactivated by heparin in the presence of plasma. In conclusion, we have developed a rapid and reproducible procedure for isolation in high yield of large quantities of the thrombin-like enzyme from the venom of the eastern diamondback rattlesnake. Studies are continuing on the primary structure and possible clinical applications of this enzyme.

Development of a Large Scale Production Process for BAX 855, a PEGylated rFVIII Product with Prolonged Half-Life

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4362-4362
Author(s):  
Jürgen Siekmann ◽  
Purtscher Martin ◽  
Oliver Zöchling ◽  
Robert Kellerer ◽  
Hanspeter Rottensteiner ◽  
...  
Keyword(s):  
Manufacturing Process ◽  
Large Scale ◽  
Specific Activity ◽  
Drug Product ◽  
Scale Production ◽  
Drug Products ◽  
Large Scale Production ◽  
Bulk Drug ◽  
Recombinant Fviii ◽  
Conjugation Process

Abstract Abstract 4362 Baxter and Nektar have developed BAX 855, a PEGylated form of Baxter’s recombinant FVIII (rFVIII) product based on the ADVATE™ manufacturing process. The conjugation process for preparing BAX 855 uses proprietary stable PEGylation from Nektar Therapeutics. Similar Nektar technology has been successfully employed for marked and licensed PEGylated drug products and drugs in clinical use. The manufacturing process for BAX 855 comprises several steps, including chromatographic purification on a MacroCap SP resin, a strong cation exchanger, which allows the fractionation of species with different PEGylation degrees and concentration of the conjugate collected by an ultra- / diafiltration step leading to the pre-formulated bulk drug substance (BDS). Final formulation of the BDS includes a filling and lyophilization step to obtain the final drug product. The process described is suited to manufacture BAX 855 in gram scale and showed a good batch to batch consistency, ensuring an equivalent product quality for each batch. BAX 855 manufactured by this process has a specific activity similar to that of rFVIII in ADVATE™ and PEGylation degrees in the narrow range of 2 to 3 mols PEG / mol rFVIII. SDS-PAGE and Western blot analysis of BAX 855 confirm the stable PEGylation and demonstrate an increase in the molecular weight of the various FVIII domains. PK studies in different species display longer survival of BAX 855 compared to ADVATE™. Disclosures: Siekmann: Baxter Innovations GmbH: Employment. Martin:Baxter Innovations GmbH: Employment. Zöchling:Baxter Innovations GmbH: Employment. Kellerer:Baxter Innovations GmbH: Employment. Rottensteiner:Baxter Innovations GmbH: Employment. Mitterer:Baxter Innovations GmbH: Employment. Bossard:Nektar Therapeutics: Employment. Phillips:Nektar Therapeutics: Employment. Ehrlich:Baxter Innovations GmbH: Employment. Scheiflinger:Baxter Innovations GmbH: Employment. Turecek:Baxter Innovations GmbH: Employment.

scholarly journals Improvement of Bacillus licheniformis MZK05 by mutation for increased production of keratinase

2015 ◽  
Vol 24 (1) ◽  
pp. 17-23 ◽  
Author(s):  
Serajus Salaheen ◽  
Md Arafat Al Mamun ◽  
Shakila Nargis Khan ◽  
Md Mozammel Hoq
Keyword(s):  
Bacillus Licheniformis ◽  
Large Scale ◽  
Skim Milk ◽  
Shake Flask Culture ◽  
Scale Production ◽  
Feather Meal ◽  
Carbon And Nitrogen ◽  
Enhanced Production ◽  
Large Scale Production ◽  
Improved Strain

Bacillus licheniformis MZK05 was subjected to mutation by ultraviolet radiation for enhanced production of keratinase. Of 750 isolates from irradiated plates, 200 colonies that showed zone of casein hydrolysis on Skim Milk Agar were cultured in liquid Feather Meal Medium containing digested feather as carbon and nitrogen source in shake culture at 37ºC. The mutant B. licheniformis MZK05M9 (BlM9) exhibited highest enzyme activity of 170 ± 5.63 U/ml as compared to 74 ± 5.29 U/ml by the wild MZK05. Both the strains were examined for the presence of gene encoded for keratinase (kerA gene) by PCR using primer which showed the product sizes 1156 bp and 520 bp, respectively for MZK05 and BlM9. The keratinase from both strains exhibited a thermal stability of about 97% for 2 hrs at 40°C whereas the keratinase of the mutant strain showed less stability (55%) at 50°C. The BlM9 while cultivated in batch culture in 7 litre bioreactor for production of the keratinase in the Feather Meal Medium, the productivity was found to be double (17,608 U/L/hr) than that of in the shake flask culture (8,525 U/L/hr). This improved strain thus will be very useful for large scale production of keratinase enabling its technical applications in industry. Dhaka Univ. J. Biol. Sci. 24(1): 17-23, 2015 (January)

scholarly journals Anti-L. donovani Activity in Macrophage/Amastigote Model of Palmarumycin CP18 and its Large Scale Production

2014 ◽  
Vol 9 (1) ◽  
pp. 1934578X1400900
Author(s):  
Humberto E. Ortega ◽  
Eliane de Morais Teixeira ◽  
Ana Rabello ◽  
Sarah Higginbotham ◽  
Luis Cubilla-Ríos
Keyword(s):  
Large Scale ◽  
Leishmania Donovani ◽  
Specific Activity ◽  
Fermentation Broth ◽  
Fungal Culture ◽  
Scale Production ◽  
Corn Meal ◽  
H Nmr ◽  
Solid Media ◽  
Large Scale Production

Palmarumycin CP18, isolated from an extract of the fermentation broth and mycelium of the Panamanian endophytic fungus Edenia sp., was previously reported with strong and specific activity against Leishmania donovani. Here we report that when the same strain was cultured on different solid media – Harrold Agar, Leonian Agar, Potato dextrose Agar (PDA), Corn Meal Agar, Honey Peptone Agar, and eight vegetables (V8) Agar – in order to determine the optimal conditions for isolation of palmarumycin CP18, no signal for this compound was observed in any of the 1H NMR spectra of fractions obtained from these extracts. However, one extract, prepared from the fungal culture in PDA contained significant amounts of CJ-12,372, a possible biosynthetic precursor of palmarumycin CP18. Edenia sp. was cultivated on a large scale on PDA and CJ-12,372 was converted to palmarumycin CP18 by oxidation of its p-hydroquinone moiety with DDQ in dioxane. Palmarumycin CP18 showed anti-leishmanial activity against L. donovani in a macrophage/amastigote model, with IC50 values of 23.5 μM.

scholarly journals Peroxidase from Coleus Forskohlii: Purification and Biochemical Characterization

2020 ◽  
Vol 5 (1) ◽  
pp. 9-20
Author(s):  
Yaaser Q. Almulaiky ◽  
Yaaser Q. Almulaiky
Keyword(s):  
Ion Exchange ◽  
Peroxidase Activity ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Gel Filtration ◽  
Catalytic Efficiency ◽  
Industrial Applications ◽  
Coleus Forskohlii ◽  
Sds Page ◽  
Optimum Ph

In this study, a peroxidase from new source was purified using ion exchange and gel filtration techniques. The recovery for peroxidase activity was 19% with 11-fold purification and specific activity of 749 unit/mg protein. Purified peroxidase demonstrated a molecular mass of 39 kDa using gel filtration and was confirmed as a single band on SDS-PAGE. The purified peroxidase revealed a broad optimum pH activity at 6.0-6.5 and 50°C temperature. The kinetic parameters for purified peroxidase toward H2O2 and guaiacol as substrates were found to be Km = 3.355, 5.395 mM, Kcat = 99.52, 79.56 s-1 and Vmax =1.531, 1.242 µmole ml-1 min-1, respectively. The catalytic efficiency (kcat/Km) of the purified peroxidase was 14.75 and 29.66 s−1 mM−1 for guaiacol and H2O2, respectively. Peroxidase activity was observed to be enhanced by Cu2+, Co2+, Ni2+ and inhibited in the presence of Sn2+, Al3+, Hg2+, NaN3, EDTA and urea. Characterization showed that peroxidase purified from C. forskohlii has the ability to be used for food industrial applications.

Synthesis of YBa2Cu3O7−x powder by autoignition of citrate-nitrate gel

1993 ◽  
Vol 8 (11) ◽  
pp. 2761-2766 ◽  
Author(s):  
Sukumar Roy ◽  
Das A. Sharma ◽  
S.N. Roy ◽  
H.S. Maiti
Keyword(s):  
Large Scale ◽  
Reduction Reaction ◽  
Ignition Process ◽  
Scale Production ◽  
Thermally Induced ◽  
Oxidation Reduction ◽  
Large Scale Production ◽  
Fine Particle Size ◽  
Oxidation Reduction Reaction ◽  
Good Potential

A simple and convenient method for the synthesis of YBa2Cu3O7−x powder is described. The technique involves autoignition of a citrate-nitrate gel resulting from a thermally induced oxidation-reduction reaction to yield an ash that upon calcination produces the desired compound. The resulting powder is pure, homogeneous, and possesses a reasonably fine particle size. The autoignition is restricted to a particular range of citrate-nitrate ratio in the gel. Attempts have been made to understand the ignition process with the help of Thermogravimetry (TG) and Differential Thermal Analysis (DTA) of the samples. The process appears to have a higher degree of reproducibility and a good potential for large-scale production.

scholarly journals Raw Glycerol as an Alternative Carbon Source for Cultivation of Exopolysaccharide-Producing Bacteria

2015 ◽  
Vol 3 (2) ◽  
pp. 61
Author(s):  
Renata Aguirre Trindade ◽  
Adriel Penha Munhoz ◽  
Carlos André Veiga Burkert
Keyword(s):  
Carbon Source ◽  
Xanthomonas Campestris ◽  
Large Scale ◽  
Carbon Sources ◽  
Environmental Benefits ◽  
The Other ◽  
Pseudomonas Oleovorans ◽  
Raw Glycerol ◽  
Biodiesel Synthesis ◽  
Alternative Carbon Source

<p>The large-scale use of biodiesel has shown significant environmental benefits as regards the reduction of global warming impacts. The increased generation of glycerol, the main byproduct of the reaction, makes necessary to propose alternatives to its use. In this context, the aim of this study was to evaluate raw glycerol (RG), a byproduct from biodiesel synthesis, as a carbon source for the cultivation of bacteria recognized as exopolysaccharides (EPSs) producers, compared with sucrose (S) and with a mixture of both components in a ratio of 1:1 w:w (SRG). The bacteria used were: <em>Xanthomonas campestris </em>pv. <em>mangiferaeindicae</em> IBSBF 1230, <em>Pseudomonas oleovorans</em> NRRL B-14683, <em>Sphingomonas capsulata</em> NRRL B-4261 and <em>Zymomonas mobilis</em> NRRL B-4286. All bacteria were capable of growing and producing EPSs using RG as the sole carbon source. For <em>X</em>. <em>campestris</em>, EPSs concentration of around 4.00 g L<sup>-1</sup> was found for the different carbon sources tested. For <em>P. oleovorans</em>, only the medium composed by S (0.85 g L<sup>-1</sup>) differed from the other media, with better results being found using RG and SRG. <em>S. capsulata</em> showed higher concentration in the medium containing S and SRG, around 3.40 g L<sup>-1</sup>, and in the medium containing RG this value decreased to 1.70 g L<sup>-1</sup>. <em>Z. mobilis</em>, on the other hand, showed a better result using SRG (1.41 g L<sup>-1</sup>), and in the medium containing S and RG, these values were lower, reaching 0.27 and 0.77 g L<sup>-1</sup>, respectively.</p>

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