Characterization of arylsulfatase isoenzymes from Pseudomonas aeruginosa

1972 ◽  
Vol 18 (5) ◽  
pp. 561-568 ◽  
Author(s):  
G. J. Delisle ◽  
F. H. Milazzo
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Kinetic Properties ◽  
Type I ◽  
Similar Kinetic ◽  
Substrate Hydrolysis ◽  
Curious Property

A method is described for the separation and isolation of two electrophoretically distinct arylsulfatases (arylsulfatase EC.3.1.6.1) from Pseudomonas aeruginosa.Characterization of the enzymes revealed that they were type I arylsulfatases, with similar kinetic properties. They differed, however, in respect to charge, pH optima for substrate hydrolysis, and activation by anions. In addition, the enzymes displayed dual pH optima curves with both substrates used and the curious property of a shift in pH optima with varied p-nitrophenyl sulfate concentrations.

Characterization of a type I pullulanase from Anoxybacillus sp. SK3-4 reveals an unusual substrate hydrolysis

2016 ◽  
Vol 100 (14) ◽  
pp. 6291-6307 ◽  
Author(s):  
Ummirul Mukminin Kahar ◽  
Chyan Leong Ng ◽  
Kok-Gan Chan ◽  
Kian Mau Goh
Keyword(s):  
Type I ◽  
Substrate Hydrolysis

scholarly journals Identification and characterization of the gene encoding the human phosphopantetheine adenylyltransferase and dephospho-CoA kinase bifunctional enzyme (CoA synthase)

2002 ◽  
Vol 365 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Suren AGHAJANIAN ◽  
D.Margaret WORRALL
Keyword(s):  
Biosynthetic Pathway ◽  
Sequence Similarity ◽  
Kinetic Properties ◽  
Sequence Information ◽  
Bifunctional Enzyme ◽  
Gene Encoding ◽  
Pig Liver ◽  
Identification And Characterization ◽  
Similar Kinetic

The final two enzymes in the CoA biosynthetic pathway, phosphopantetheine adenylyltransferase (PPAT; EC 2.7.7.3) and dephospho-CoA kinase (DPCK; EC 2.7.1.24), are separate proteins in prokaryotes, but exist as a bifunctional enzyme in pig liver. In the present study we have obtained sequence information from purified pig-liver enzyme, and identified the corresponding cDNA in a number of species. The human gene localizes to chromosome 17q12-21 and contains regions with sequence similarity to the monofunctional Escherichia coli DPCK and PPAT. The recombinant 564-amino-acid human protein confirmed the associated transferase and kinase activities, and gave similar kinetic properties to the wild-type pig enzyme.

scholarly journals Isolation and characterization of pigeon breast muscle cytosolic 5'-nucleotidase-I (cN-I).

2005 ◽  
Vol 52 (4) ◽  
pp. 789-796 ◽  
Author(s):  
Kinga Tkacz-Stachowska ◽  
Katarzyna Lechward ◽  
Andrzej C Skladanowski
Keyword(s):  
Physiological Function ◽  
Specific Activity ◽  
Heart Tissue ◽  
Breast Muscle ◽  
Kinetic Properties ◽  
Type I ◽  
Isolation And Characterization ◽  
Atp Breakdown ◽  
Wet Weight

5'-Nucleotidase specific towards dCMP and AMP was isolated from avian breast muscle and characterized. It was found to be similar to a type-I form (cN-I) identified earlier as the AMP-selective 5'-nucleotidase responsible for adenosine formation during ATP breakdown in transfected COS-7 cells. Expression pattern of the cN-I gene in pigeon tissues indicated breast muscle as a rich source of the transcript. We purified the enzyme from this source using two-step chromatography and obtained an active homogenous preparation, free of ecto-5'-nucleotidase activity. The tissue content of the activity was calculated at 0.09 U/g wet weight. The specific activity of the enzyme preparation was 4.33 U/mg protein and it preferred dCMP and AMP to dAMP and IMP as a substrate. Its kinetic properties were very similar to those of the enzyme purified earlier from heart tissue. It was strongly activated by ADP. Inhibition by inorganic phosphate was more pronounced than in heart-isolated cN-I. Despite this difference, a similar physiological function is suggested for cN-I in both types of muscle.

scholarly journals Genetic and biochemical characterization of the Na + /H + antiporters of Pseudomonas aeruginosa

2021 ◽  
Author(s):  
Sara Foreman ◽  
Kristina Ferrara ◽  
Teri Hreha ◽  
Ana Duran-Pinedo ◽  
Jorge Frias-Lopez ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biochemical Characterization ◽  
Physiological Role ◽  
Glycerol Metabolism ◽  
Kinetic Properties ◽  
Wild Type ◽  
Wide Range ◽  
Quadruple Mutant

Pseudomonas aeruginosa has four Na + /H + antiporters that interconvert and balance Na + and H + gradients across the membrane. These gradients are important for bioenergetics and ionic homeostasis. To understand these transporters, we have constructed four strains, each of which has only one antiporter: NhaB, NhaP, NhaP2, and Mrp. We also constructed a quadruple deletion mutant that has no Na + /H + antiporters. Although the antiporters of P. aeruginosa have previously been studied, the strains constructed here present the opportunity to characterize their kinetic properties in their native membranes and their roles in the physiology of P. aeruginosa . The strains expressing only NhaB or Mrp, the two electrogenic antiporters, are able to grow essentially as the wild type across a range of [Na + ] and pH. Strains with only NhaP or NhaP2, which are electroneutral, grow more poorly at increasing [Na + ], especially at high pH, with NhaP the most sensitive. The strain with no Na + /H + antiporters is extremely sensitive to [Na + ] and shows essentially no Na + (Li + )/H + antiporter activity but retains most K + /H + antiporter activity of the wild type at pH 7.5 and approximately half at pH 8.5. We also used the four strains that each express one of the four antiporters to characterize the kinetic properties of each transporter. RNA-seq analysis of the quadruple deletion strain showed widespread changes, including pyocyanin synthesis, biofilm formation, and nitrate and glycerol metabolism. Thus, the strains constructed for this study will open a new door to understanding the physiological role of these proteins and their activities in P. aeruginosa . Importance Pseudomonas aeruginosa has four Na + /H + antiporters that connect and interconvert its Na + and H + gradients. We have constructed four deletion mutants, each of which has only one of the four Na + /H + antiporters. These strains made it possible to study the properties and physiological roles of each antiporter independently in its native membrane. Mrp and NhaB are each able to sustain growth over a wide range of pH and [Na + ], whereas the two electroneutral antiporters, NhaP and NhaP2, are most effective at low pH. We also constructed a quadruple mutant, lacking all four antiporters in which the H + and Na + gradients are disconnected. This will make it possible to study the role of the two gradients independently.

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