Pseudomonas fluorescens extraction

In Pseudomonas fluorescens extract, histidase free from urocanase can be prepared by heating the extract at 85°C. for 15 minutes. By this procedure all ability to destroy urocanic acid is lost, but the full capacity to deaminate histidine is retained when the product is supplemented with 10 M glutathione. 

It is nearly 50 years since a c-type cytochrome was shown to catalyze peroxidase activity in crude extracts of Pseudomonas fluorescens (40). The enzyme responsible was first purified some 20 years later by Ell-folk and Soninen from the closely related P. aeruginosa and shown to be a diheme cytochrome c peroxidase (CCP) (41). These bacterial diheme CCPs are quite distinct from the superfamily of plant and yeast peroxidases (42) and are widely distributed among the Gram-negative bacteria (41, 43 6). Diheme CCPs are located in the periplasm (Fig. 2), where they catalyze the two-electron reduction of H2O2 to H2O by soluble one-electron donors such as cytochromes c and cupredoxins. 

M. phlei, and several strains of M. tuberculosis (28-30) contain a true asparaginase, as do extracts of Bacillus coagulans and Bacillus stearo-thermophilus (31). Brucella abortus contains two asparaginases, one specific for L-asparagine and the other for the opposite enantiomorph (32). An asparaginase is also present in Pseudomonas fluorescens (33). Also, Tsuji, in 1957, had reported the presence of asparaginase in extracts of acetone powders of E. coli, Staphylococcus, M. avium, and Aspergillus oryzae (34). 

Marin, A., Mawhinney, T.P., Marshall, R.T. 1984. Glycosidic activities of Pseudomonas fluorescens on fat-extracted skim milk, buttermilk and milk fat globule membranes. J. Dairy Sci. 67, 52-59. 

Petrovic et al. [37] investigated the antibacterial activity of the water, ethanol and ethyl acetate extracts of Cichorium intybus L. Water extract inhibited Agrobacterium radiobacter spp. tumefaciens, Erwinia carotovora. Pseudomonas fluorescens and Pseudomonas aeruginosa. The plant also showed antifungal activity against phytopathogenic fungi [28]. 

Investigations by Colowick et al. (1) on isocitrate dehydrogenase in Pseudomonas fluorescens led to the discovery that in the presence of extracts of these bacteria NAD+ could be reduced by isocitrate provided a catalytic amount of NADP+ was added. It was proposed that a specific enzyme, called pyridine nucleotide transhydrogenase, catalyzed the for-... 

More recently acids have been resolved with enzymes cloned and over-expressed in their own organisms, such as an esterase from Bacillus subtilis that resolves ibuprofen methyl ester 138 to give ibuprofen 139 of 99% ee. A range of anti-inflammatory arylpropionic esters, including 138, could also be resolved with a cell-free extract from Pseudomonas fluorescens showing that purified enzymes are not essential.34... 

Typical kinetic resolutions of the arylpropionic acids are those of flurbiprofen 16 and ketoprofen 18 with a cell-free extract of Pseudomonas fluorescens. Note the special ester (trifluoroethyl) selected for maximum efficiency and how successful that is perfect ee in the hydrolysed products at close to 50% conversion.8 The unreacted esters 15 and 17 can of course be racemised by enolisation and added to the next resolution. 

The range of phosphonic acids which can be utilized enzymically (at least by those enzymes present in Pseudomonas fluorescens) has recently been extended to include aminoalkyl-, hydroxyalkyl- and oxoalkyl-phosphonic acids, as well as phosphonodipep-tides only slight growth was seen with the herbicide glyphosate (A-phosphonomethyl-glycine (83), which is metabolized to (aminomethyl)phosphonic acid, and neither phos-phinothricin (84) nor its dialanyl tripeptide bialaphos supports growth in contrast to the behaviour of other bacterial extracts isolated from soil ". ... 

Dihydroxybenzoate is oxidized by extracts of Pseudomonas fluorescens with the consumption of one mole of oxygen per mole of substrate. An equivalent amount of carbon dioxide is evolved, and a-hydroxymuconic semialdehyde is formed. Intermediates between 2,3-dihydroxyben-zoate and a-hydroxymuconic semialdehyde have not been detected, nor has the site of ring cleavage been established. 

Figure 1. 2,3-Dihydroxybenzoate oxidation by partially purified extracts of Pseudomonas fluorescens...

Viscosinamide, a cydodepsipeptide — depsipeptides) isolated from an extract of Pseudomonas fluorescens DR54. It contains Leu (3), d-G1u, D-flHo-Thr, D-Val, D-Ser (2), He, and -hydroxydecanoic acid with the ester bond between D-allo-Thr and Ile. Viscosinamide exhibits antifungal and surfactant properties [T. H. Nielsen et al, J. Appl. Microbiol. 1999, 87, 80]. 

Odham et al. (1988) compared plasmaspray with thermospray by using ammonium acetate as a buffer, where PSP required no addition of buffer prior to ionization. The total ion current obtained in PSP was significantly higher than previously reported for TS (filament-on). No or very few molecular ions were observed as a result of the harder ionization in PSP compared with TS. Diacylglycerol- and monoacylglycerol-derived fragments of phospholipids were obtained. Cation-exchange HPLC separated PI and PE in a phospholipid extract from bacterial cells Pseudomonas fluorescens). Furthermore, PC, PE, PA, PS, cardiolipin (CL) and polyphosphoinositides (PIP and PIP-2) were also studied. 

Valeur, Michelsen and Odham (1993) showed how diacylglycerol- and monoacylglycerol-derived fragments of phospholipids were obtained. PG and PE were found in a phospholipid extract from bacterial cells Pseudomonas fluorescens). They achieved class separation of the phospholipids but did not apparently separate molecular species (Fig. 9.6). Collision-induced dissociation (CID)-MS-MS analyses of PI from soybean were performed which revealed specific fatty acid compositions of the selected diacylglycerol-derived fragment. 

The correct catabolic pattern of histidine was established first in bacterial preparations and for this reason will be discussed first. Cell-free extracts of Pseudomonas fluorescens were obtained which catalyze the quantitative conversion of L-histidine to L-glutamic acid, formic acid, and 2 moles of NH3, as required by equation 14. Proof of the formation of L-glutamate was obtained by isolating the crystalline hydrochloride from the incubation mixture and establishing its identity. ... 

Electron Transfer. Another mechanism for the interaction of the pyridine nucleotides was described by Colowick et al. They observed an enzyme in extracts of Pseudomonas fluorescens, Azotobacter and, more... 

This enzyme was discovered by Hayaishi and Hashimoto 364), ft has been purified and studied from acetone-dried Micrococcus ureae cells (366) and from tryptophan-adapted Pseudomonas fluorescens cell extracts (366). The enzyme has a molecular weight of about 80,000 and contains... 

A number of heteroaromatic monothiocarboxylic acids are formed by Pseudomonas sp. From P. putida, there was isolated pyridine-2,6-di-(mon-othiocarboxylic acid) 46 (Scheme 16). Of interest is the fact that in P. stutzeri KC, a copper complex of 46 is the active agent for a one electron transfer in the bacterial biodegradation of CCI4. Methylation of P. putida extracts provides a number of related structures such as 47. In addition, a P. fluorescens sp. contains 8-hydroxy-4-methoxy-quinoline-2-monothiocarboxylic acid 48.98... 

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