Utilization of triaryl phosphates by a mixed bacterial population

1975 ◽  
Vol 21 (2) ◽  
pp. 140-145 ◽  
Author(s):  
M. A. Pickard ◽  
J. A. Whelihan ◽  
D. W. S. Westlake
Keyword(s):  
Inorganic Phosphate ◽  
Major Part ◽  
Culture Medium ◽  
Bacterial Population ◽  
Extracellular Enzymes ◽  
Culture Supernatant ◽  
Enrichment Culture ◽  
Mixed Population ◽  
Triphenyl Phosphate ◽  
Cell Numbers

A mixed bacterial population that has been isolated by enrichment culture is capable of growth on Fyrquel 220, a commercial triaryl phosphate lubricant, as sole carbon source.The mixture was dominated by a yellow, Gram-negative rod which made up greater than 60% of the mixture. However, all attempts to grow this organism in pure culture on triaryl phosphate were unsuccessful. The mixed population was also capable of growth on tri-o-cresyl phosphate, trixylenyl phosphate, and triphenyl phosphate as sole carbon sources.Viable cell numbers increased 20- to 30-fold, reaching a maximum after 72–96 h growth. Only a small portion of the triaryl phosphate was used for growth; the major part was emulsified and remained in the culture medium. No evidence of extracellular enzymes capable of triaryl phosphate degradation could be found in concentrates of the culture supernatant after growth, though traces of what may have been triaryl phosphate breakdown products were observed.Cell-free extracts of the mixed culture catalyzed the release of inorganic phosphate when incubated with Fyrquel 220, tri-o-cresyl phosphate, trixylenyl phosphate, or triphenyl phosphate, indicating the presence of a phosphotriesterase or of a phosphodiesterase of wide specificity.

Production and control of extracellular enzymes in Micrococcus sodonensis

1974 ◽  
Vol 20 (1) ◽  
pp. 81-90 ◽  
Author(s):  
Cecily Mills ◽  
J. N. Campbell
Keyword(s):  
Alkaline Phosphatase ◽  
Inorganic Phosphate ◽  
Culture Medium ◽  
Extracellular Enzymes ◽  
Synthetic Medium ◽  
Culture Conditions ◽  
The Other ◽  
Organic Nutrients ◽  
And Control ◽  
Logarithmic Growth

Micrococcus sodonensis has been shown to produce several extracellular enzymes: an alkaline phosphatase, at least two forms of phosphodiesterase, a 5′-nucleotidase, and an alkaline proteinase. The quantitative release of these enzymes into the culture medium during logarithmic growth under all the various culture conditions tested indicates that these enzymes are truly extracellular in nature. Inorganic phosphate repressed the production of the alkaline phosphatase in synthetic as well as in complex media, whereas, the repression of the production of active diesterase and 5′-nucleotidase by inorganic phosphate was partly reversed by the addition of supplemental organic nutrients to the culture medium. Proteinase production was independent of the culture conditions used. A mutant strain of M. sodonensis with an altered production of diesterase was obtained; the other extracellular enzymes were unaffected. These results suggest that the extracellular enzymes of M. sodonensis are not produced in a pleiotropic fashion since the level of one of the enzymes can be changed without affecting a corresponding change in the levels of the other enzymes. An extracellular high molecular weight carbohydrate fraction was shown to be produced by M. sodonensis in synthetic medium. The fraction was also shown to contain glycoprotein.

scholarly journals Characterization and optimization of extracellular enzymes production by Aspergillus niger strains isolated from date by-products

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Reda Bellaouchi ◽  
Houssam Abouloifa ◽  
Yahya Rokni ◽  
Amina Hasnaoui ◽  
Nabil Ghabbour ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Culture Medium ◽  
Extracellular Enzymes ◽  
Enzyme Expression ◽  
Specific Enzyme ◽  
Enzymatic Production ◽  
High Production Rate ◽  
High Production ◽  
Incubation Temperatures ◽  
By Products

Abstract Background This work aims to study the optimal conditions of the fermentation culture medium used for the production of extracellular enzymes (amylase, cellulase, lipase, and protease) from previously isolated Aspergillus niger strains in date by-products. Results The five most powerful isolates selected based on the zone of degradation formed on Petri plates by the substrate were subjected to the quantitative evaluation of their enzymatic production. All five strains showed almost similar API-ZYM profiles, with minor variations observed at the level of some specific enzyme expression. The production of cellulase and amylase was depending on pH and incubation temperatures. ASP2 strain demonstrated the high production rate of amylase (at pH 5 and 30 °C) and cellulase (at pH 6 and 30 °C) for 96 h of incubation. Conclusion The A. niger showed the ability to produce several extracellular enzymes and can be used in the valorization of different agroindustrial residues.

scholarly journals Optimisation of xylanases production by two Cellulomonas strains and their use for biomass deconstruction

2021 ◽  
Author(s):  
Ornella M Ontañon ◽  
Soma Bedő ◽  
Silvina Ghio ◽  
Mercedes M Garrido ◽  
Juliana Topalian ◽  
...  
Keyword(s):  
Extracellular Enzymes ◽  
Culture Supernatant ◽  
Xylanase Activity ◽  
Barley Straw ◽  
Proteomic Profiling ◽  
Biomass Hydrolysis ◽  
Biomass Deconstruction ◽  
Xylanolytic Activity ◽  
Key Points ◽  
Distinguishing Features

Abstract One of the main distinguishing features of bacteria belonging to the Cellulomonas genus is their ability to secrete multiple polysaccharide degrading enzymes. However, their application in biomass deconstruction still constitutes a challenge. We addressed the optimisation of the xylanolytic activities in extracellular enzymatic extracts of Cellulomonas sp. B6 and Cellulomonas fimi B-402 for their subsequent application in lignocellulosic biomass hydrolysis by culture in several substrates. As demonstrated by secretomic profiling, wheat bran and waste paper resulted to be suitable inducers for the secretion of xylanases of Cellulomonas sp. B6 and C. fimi B-402, respectively. Both strains showed high xylanolytic activity in culture supernatant although Cellulomonas sp. B6 was the most efficient xylanolytic strain. Upscaling from flasks to fermentation in a bench scale bioreactor resulted in equivalent production of extracellular xylanolytic enzymatic extracts and freeze drying was a successful method for concentration and conservation of the extracellular enzymes, retaining 80% activity. Moreover, enzymatic cocktails composed of combined extra and intracellular extracts effectively hydrolysed the hemicellulose fraction of extruded barley straw into xylose and xylooligosaccharides. Key points • Secreted xylanase activity of Cellulomonas sp. B6 and C. fimi was maximised. • Biomass-induced extracellular enzymes were identified by proteomic profiling. • Combinations of extra and intracellular extracts were used for barley straw hydrolysis.

scholarly journals EXPRESSION OF b-XYLOSIDASE ENCODING GENE IN PHIS/ Bacillus megaterium MS SYSTEM

2015 ◽  
Vol 2 (1) ◽  
pp. 25 ◽  
Author(s):  
Sri Sumarsih
Keyword(s):  
Culture Medium ◽  
Culture Supernatant ◽  
Specific Activity ◽  
Agar Medium ◽  
Nitrilotriacetic Acid ◽  
Protoplast Transformation ◽  
E Coli ◽  
Recombinant Cells ◽  
Relative Molecular Weight ◽  
Encoding Gene

b-Xylosidase encoding gene from G. thermoleovorans IT-08 had been expressed in the pHIS1525/ B. megaterium MS941 system. The b-xylosidase gene (xyl) was inserted into plasmid pHIS1525 and propagated in E. coli DH10b. The recombinant plasmid was transformed into B. megaterium MS941 by protoplast transformation. Transformants were selected by growing the recombinant cells on solid LB medium containing tetracycline (10 µg/ ml). The expression of the b-xylosidase gene was assayed by overlaid the recombinant B. megaterium MS941 cell with agar medium containing 0.2% ethylumbelliferyl-b-D-xyloside (MUX). This research showed that the b-xylosidase gene was succesfully sub-cloned in pHIS1525 system and expressed by the recombinant B. megaterium MS941. Theaddition of 0.5% xylose into the culture medium could increase the activity of recombinantactivity of recombinant of recombinantb-xylosidase by 2.74 fold. The recombinant B. megaterium MS941 secreted 75.56% of the expressed b-xylosidase into culture medium. The crude extract b-xylosidase showed the optimum activity at 50° C and pH 6. The recombinant b-xylosidase was purified from culture supernatant by affinity chromatographic method using agarose containing Ni-NTA (Nickel-Nitrilotriacetic acid). The pure b-xylosidase showed a specific activity of 10.06 Unit/mg protein and relative molecular weight ± 58 kDa.

scholarly journals Enzyme Bioprospection of Marine-Derived Actinobacteria from the Chilean Coast and New Insight in the Mechanism of Keratin Degradation in Streptomyces sp. G11C

Marine Drugs ◽  
2020 ◽  
Vol 18 (11) ◽  
pp. 537
Author(s):  
Valentina González ◽  
María José Vargas-Straube ◽  
Walter O. Beys-da-Silva ◽  
Lucélia Santi ◽  
Pedro Valencia ◽  
...  
Keyword(s):  
Substrate Concentration ◽  
Major Part ◽  
Extracellular Enzymes ◽  
Incubation Temperature ◽  
Inoculum Size ◽  
Marine Actinobacteria ◽  
Valuable Alternative ◽  
Streptomyces Sp ◽  
Abundance And Diversity ◽  
Promising Source

Marine actinobacteria are viewed as a promising source of enzymes with potential technological applications. They contribute to the turnover of complex biopolymers, such as pectin, lignocellulose, chitin, and keratin, being able to secrete a wide variety of extracellular enzymes. Among these, keratinases are a valuable alternative for recycling keratin-rich waste, which is generated in large quantities by the poultry industry. In this work, we explored the biocatalytic potential of 75 marine-derived actinobacterial strains, focusing mainly on the search for keratinases. A major part of the strains secreted industrially important enzymes, such as proteases, lipases, cellulases, amylases, and keratinases. Among these, we identified two streptomycete strains that presented great potential for recycling keratin wastes—Streptomyces sp. CHA1 and Streptomyces sp. G11C. Substrate concentration, incubation temperature, and, to a lesser extent, inoculum size were found to be important parameters that influenced the production of keratinolytic enzymes in both strains. In addition, proteomic analysis of culture broths from Streptomyces sp. G11C on turkey feathers showed a high abundance and diversity of peptidases, belonging mainly to the serine and metallo-superfamilies. Two proteases from families S08 and M06 were highly expressed. These results contributed to elucidate the mechanism of keratin degradation mediated by streptomycetes.

The metabolism of p-fluorophenylacetic acid by a Pseudomonas sp. I. Isolation and identification of intermediates in degradation

1971 ◽  
Vol 17 (5) ◽  
pp. 635-644 ◽  
Author(s):  
D. B. Harper ◽  
E. R. Blakley
Keyword(s):  
Sole Carbon Source ◽  
Culture Medium ◽  
Enrichment Culture ◽  
Culture Technique ◽  
Spectroscopic Techniques ◽  
Resting Cells ◽  
Pseudomonas Sp ◽  
Isolation And Identification ◽  
Hydroxyphenylacetic Acid ◽  
Organic Fluorine

A Pseudomonas sp. capable of growing on p-fluorophenylacetic acid as sole carbon source has been isolated using the enrichment culture technique. All the organic fluorine is released into the culture medium as fluoride ion during growth. A number of fluorinated intermediates have been isolated from the culture medium when resting cells were incubated with the substrate. Using infrared, nuclear magnetic resonance, and mass spectroscopic techniques together with chemical degradative procedures, these have been identified as D(+)-monofluorosuccinic acid, trans-3-fluoro-3-hexenedioic acid, (−)-4-carboxymethyl-4-fluorobutanolide, 4-fluoro-2-hydroxyphenylacetic acid, and 4-fluoro-3-hydroxyphenylacetic acid.

scholarly journals Development of an optimized broth enrichment culture medium for the isolation of Clostridium difficile

Anaerobe ◽  
2018 ◽  
Vol 54 ◽  
pp. 92-99 ◽  
Author(s):  
Mairéad C. Connor ◽  
John W. McGrath ◽  
Geoff McMullan ◽  
Nikki Marks ◽  
Derek J. Fairley
Keyword(s):  
Clostridium Difficile ◽  
Culture Medium ◽  
Enrichment Culture ◽  
Broth Enrichment

286 INFLUENCE OF REVERSIBLE MEIOSIS INHIBITION ON SWINE EMBRYOS PRODUCED BY IN VITRO FERTILIZATION AND PATHENOGENETIC ACTIVATION

2006 ◽  
Vol 18 (2) ◽  
pp. 250
Author(s):  
M. G. Marques ◽  
A. B. Nascimento ◽  
V. P. Oliveira ◽  
A. R. S. Coutinho ◽  
M. E. O. A. Assumpção ◽  
...  
Keyword(s):  
Culture Medium ◽  
Cell Number ◽  
Control Group ◽  
Cleavage Rate ◽  
Vitro Fertilization ◽  
Electric Pulses ◽  
Cell Numbers ◽  
Blastocyst Rate ◽  
And Control

The present work evaluated the reversible meiosis inhibition effect on the development of swine embryos produced by in vitro fertilization (IVF) or parthenogenetic activation (PA). The efficiency of PZM3 and NCSU23 embryo culture media was also evaluated. Oocytes from ovaries collected at a slaughterhouse were subjected to IVM in two different groups: CHX (cycloheximide 5 µM for 10 h) and control, both with TCM-199 + 3.05 mM glucose + 0.91 mM sodium pyruvate + 10% porcine follicular fluid (pFF) + 0.57 mM cystein + 10 ng epidermal growth factor (EGF)/mL + 10 IU eCG/mL + 10 IU hCG/mL for the initial 22 h. In the remaining period (20 h for CHX and 22 h for control), medium without hormones was utilized. After IVM, oocytes were denuded and fertilized for 6 h (IFV) or the matured oocytes were submitted to activation by electric pulses (PA) (2 DC of 1.5 kV/cm for 30 µs), incubated for 1 h in culture medium with 10 μM of CHX, and again submitted to the same electric pulses for 60 µs. Embryo development was evaluated by cleavage rate on Day 3 and blastocyst rate and blastocyst cell number on Day 7 of culture. Cleavage and blastocyst rates were analyzed by the equality-of-two-ratios test and cell number by the Kruskal-Wallis and Mann-Whitney tests (P < 0.05). In relation to IVF, the PZM3 medium was more efficient than NCSU23 for cleavage rate in the CHX group (PZM3: 68.4%, NCSU23: 44.4%) and had a better blastocyst rate in the control group (PZM3: 13.4%, NCSU23: 5.6%). With reference to PA, NCSU23 presented better cleavage and blastocyst rates than PZM3 in the CHX group (NCSU23: 89.5%, PZM3: 78.5% and NCSU23: 20.4%, PZM3: 13.0%, respectively). In the control group, only the NCSU23 blastocyst rate was higher than that for PZM3 (NCSU23: 22.5%, PZM3: 10.8%). No culture medium effect on cell number mean of IVF and PA blastocysts was observed. Maturation block improved cleavage rates in IVF groups cultured with PZM3 (68.4% and 50.6%, respectively, for CHX and control) and in PA groups cultured with NCSU23 (89.5% and 80.3%, respectively, for CHX and control), but no improvement of blastocyst rates in both groups (IVF and PA) was verified. Table 1 below shows that maturation block decreased the IVF and increased the PA blastocyst cell numbers. As older oocytes are more effectively activated, oocytes blocked with CHX achieved the maturation stage faster than the control group, therefore resulting in high-quality PA blastocysts. In conclusion, PZM3 was more efficient for IVF embryo production in contrast to NCSU23, whereas NCSU23 can be indicated for PA embryo production. Moreover, maturation blockage with CHX influenced blastocyst cell number, decreasing in IVF embryos and increasing in PA embryos. Table 1. Mean (±SD) of blastocyst cell numbers for IVF or PA groups after in vitro maturation without (control) or with cycloheximide (CHX) and cultured in NCSU23 or PZM3 medium This work was supported by FAPESP 02/10747–1.

RESPIRATION AND PHOSPHORUS-CONTAINING COMPOUNDS IN NORMAL AND TUMOR TISSUES OF RED BEET ROOTS

1962 ◽  
Vol 40 (9) ◽  
pp. 1251-1256 ◽  
Author(s):  
K. J. Scott ◽  
R. M. Smillie ◽  
G. Krotkov
Keyword(s):  
Inorganic Phosphate ◽  
Analytical Study ◽  
The Other ◽  
Cell Numbers ◽  
Normal Tissues ◽  
Red Beet ◽  
Tumor Tissues ◽  
Labile Phosphorus ◽  
Phosphorus Containing ◽  
Acid Labile

A comparative analytical study has been made of normal and tumor tissues of red beet roots. Per μg DNA, the two tissues had similar cell numbers, protein contents, and respiratory rates. On the other hand, the fresh and dry weights, acid-soluble phosphorus including nucleotides, acid-labile phosphorus, and inorganic phosphate were higher in the normal tissues. Tumor tissues contained more RNA. These results are discussed in relation to the cellular metabolism of the normal and tumor tissues and it is suggested that the main metabolic differences between these tissues lie in the utilization of respiratory energy, rather than in its production.

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