Document Detail


Localization of alkaline phosphatase in three gram-negative rumen bacteria.
MedLine Citation:
PMID:  4147649     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Of the three species (Bacteroides ruminicola, B. succinogenes, and Megasphaera elsdenii) of anaerobic gram-negative rumen bacteria studied, only B. ruminicola produced significant amounts of alkaline phosphatase. This enzyme, which is constitutive, showed a greater affinity for p-nitrophenylphosphate than for sodium-beta-glycerophosphate and was shown to be located exclusively in the periplasmic space of log-phase cells. Small amounts of this enzyme were released from these cells in stationary-phase cultures, but washing in 0.01 M MgCl(2) and the production of spheroplasts by using lysozyme in 0.01 M MgCl(2) did not release significant amounts of the enzyme. Exposure to 0.2 M MgCl(2) did not release significant amounts of the periplasmic alkaline phosphatase of the cell, and when these cells were spheroplasted with lysozyme in 0.2 M MgCl(2) only 25% of the enzyme was released. Spheroplasts were formed spontaneously in aging cultures of B. ruminicola, but even these cells retained most of their periplasmic alkaline phosphatase. It was concluded that the alkaline phosphatase of B. ruminicola is firmly bound to a structural component within the periplasmic area of the cell wall and that the enzyme is released in large amounts only when the cells break down. The behavior of alkaline phosphatase in this bacterium contrasts with that of conventional periplasmic enzymes of aerobic bacteria, which are released upon conversion into spheroplasts by lysozyme and ethylenediaminetetraacetic acid and by other types of cell wall damage. All three species of bacteria studied here, as well as bacteria found in mixed populations in the rumen, have thick, complex layers external to the double-track layer of their cell walls. In addition, B. ruminicola produces a loose extracellular material.
Authors:
K J Cheng; J W Costerton
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of bacteriology     Volume:  116     ISSN:  0021-9193     ISO Abbreviation:  J. Bacteriol.     Publication Date:  1973 Oct 
Date Detail:
Created Date:  1973-12-16     Completed Date:  1973-12-16     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2985120R     Medline TA:  J Bacteriol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  424-40     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / biosynthesis,  isolation & purification*,  metabolism
Anaerobiosis
Animals
Bacteria / enzymology*
Bacteroides / enzymology*
Cell Wall / enzymology
Cell-Free System
Glutamate Dehydrogenase / metabolism
Magnesium
Microscopy, Electron
Microscopy, Phase-Contrast
Muramidase
NADH, NADPH Oxidoreductases / metabolism
Rumen / microbiology*
Species Specificity
Spectrophotometry
Spheroplasts / enzymology
Vibration
Chemical
Reg. No./Substance:
7439-95-4/Magnesium; EC 1.4.1.2/Glutamate Dehydrogenase; EC 1.6.-/NADH, NADPH Oxidoreductases; EC 3.1.3.1/Alkaline Phosphatase; EC 3.2.1.17/Muramidase
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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