HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lee, Y.-P. Articles by Erwin, M. J. Search for Related Content PubMed PubMed Citation Articles by Lee, Y.-P. Articles by Erwin, M. J. Social Bookmarking                      What's this?

Sugar Phosphate Phosphohydrolase

I. SUBSTRATE SPECIFICITY, INTRACELLULAR LOCALIZATION, AND PURIFICATION FROM NEISSERIA MENINGITIDIS

From the ¹ From the Department of Biochemistry, Guy and Bertha Research Laboratory, University of North Dakota School of Medicine, Grand Forks, North Dakota 58201
² From the Institute for Enzyme Research, University of Wisconsin, Madison, Wisconsin 53706

A new type of phosphohydrolase that can hydrolyze only phosphate esters of sugars was purified from Neisseria meningitidis. The -d-aldohexose 1-phosphates and d-ketohexose 1-phosphatase served as the best substrates for the sugar phosphate phosphohydrolase. The d-aldohexose 6-phosphates and d-ketohexose 6-phosphates were hydrolyzed at the same rate as the -d-hexose 1-phosphates, with the exception of d-galactose-6-P and d-mannose-6-P, which followed at 30% and 28.5%, respectively. If the phosphate group of the d-aldohexose 1-phosphates was esterified in the ß configuration at the first carbon atom, they no longer served as substrates. Both phosphate ester bonds of fructose-1,6-di-P were hydrolyzed. When position 6 of -d-glucose-1-P was in the form of a carboxyl group giving the sugar acid -d-glucuronic acid-1-P, its rate of hydrolysis was reduced only 25%. However, when position 1 of d-glucose-6-P or d-glyceraldehyde-3-P was in the form of a carboxyl group giving the nonsugars 6-P-gluconate and 3-P-glycerate, no significant hydrolysis was observed. The d-pentose phosphates and triose phosphates were hydrolyzed at a slower rate than the d-hexose 1-phosphates. The phosphate esters of the disaccharides -gentiobiose-1-P and -lactose-1-P were hydrolyzed at 66% and 15% of that of -d-glucose-1-P, respectively. Phosphate esters other than the sugar phosphates were not hydrolyzed significantly by the purified enzyme.

When cells were cultured in a complete basal medium, most of the sugar phosphate phosphohydrolase activity was specifically released into the sucrose-Tris-ethylenediaminetetraacetate and "cold water wash" fractions, implying a surface localization. When cells were maintained in a salt medium lacking glucose and lactalbumin hydrolysate, most of the activity remained with the insoluble cytoplasmic membrane fraction. The latter activity could be partially released after storing the cells at 1–3° for 3 or more weeks.

A method is described for the purification of the sugar phosphate phosphohydrolase from N. meningitidis. A similar type of enzyme was also purified from Escherichia coli.

A purified preparation of the sugar phosphate phosphohydrolase moved as a single peak in sedimentation, and the s_20,w was calculated to be 3.29 x 10^-13 sec.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Z. He, T. S. Griffin, and C. W. Honeycutt Enzymatic Hydrolysis of Organic Phosphorus in Swine Manure and Soil J. Environ. Qual., January 1, 2004; 33(1): 367 - 372. [Abstract] [Full Text] [PDF]