Linked Life Data

8361351

Source:http://linkedlifedata.com/resource/pubmed/id/8361351

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
1993-9-30
pubmed:abstractText
The alkane hydroxylase system of Pseudomonas oleovorans, which catalyses the initial oxidation of aliphatic substrates, is encoded by three genes. One of the gene products, the alkane hydroxylase AlkB, is an integral cytoplasmic membrane protein. Induction leads to the synthesis of 1.5-2% AlkB relative to the total cell protein, both in P. oleovorans and in recombinant Escherichia coli DH1. We present a study on the induction and localization of the alkane hydroxylase in E. coli W3110, which appears to be an interesting host strain because it permits expression levels of AlkB of up to 10-15% of the total cell protein. This expression level had negative effects on cell growth. The phospholipid content of such cells was about threefold higher than that of wild-type W3110. Freeze-fracture electron microscopy showed that induction of the alk genes led to the appearance of membrane vesicles in the cytoplasm; these occurred much more frequently in cells expressing alkB than in the negative control, which contained all of the alk genes except for alkB. Isolation and separation of the membranes of cells expressing alkB by density gradient centrifugation showed the customary cytoplasmic and outer membranes, as well as a low-density membrane fraction. This additional fraction was highly enriched in AlkB, as shown both by SDS-PAGE and enzyme activity measurements. A typical cytoplasmic membrane protein, NADH oxidase, was absent from the low-density membrane fraction. alkB expression in W3110 changed the composition of the phospholipid headgroup in the membrane, as well as the fatty acid composition of the membrane. The major changes occurred in the unsaturated fatty acids: C16:1 and C18:1 increased at the expense of C17:0cyc and C19:0cyc.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0950-382X
pubmed:author
pubmed:issnType
Print
pubmed:volume
8
pubmed:geneSymbol
alk, alkF, alkG, alkH, alkJ, alkK, alkL
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1039-51
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:8361351-Alkane 1-Monooxygenase, pubmed-meshheading:8361351-Alkanes, pubmed-meshheading:8361351-Bacterial Proteins, pubmed-meshheading:8361351-Cyclopropanes, pubmed-meshheading:8361351-Cytochrome P-450 Enzyme System, pubmed-meshheading:8361351-Enzyme Induction, pubmed-meshheading:8361351-Escherichia coli, pubmed-meshheading:8361351-Freeze Fracturing, pubmed-meshheading:8361351-Genes, Bacterial, pubmed-meshheading:8361351-Intracellular Membranes, pubmed-meshheading:8361351-Membrane Proteins, pubmed-meshheading:8361351-Mixed Function Oxygenases, pubmed-meshheading:8361351-Oxidation-Reduction, pubmed-meshheading:8361351-Phospholipids, pubmed-meshheading:8361351-Pseudomonas, pubmed-meshheading:8361351-Recombination, Genetic, pubmed-meshheading:8361351-Transformation, Bacterial
pubmed:year
1993
pubmed:articleTitle
The alkane oxidation system of Pseudomonas oleovorans: induction of the alk genes in Escherichia coli W3110 (pGEc47) affects membrane biogenesis and results in overexpression of alkane hydroxylase in a distinct cytoplasmic membrane subfraction.
pubmed:affiliation
Institute for Biotechnology, ETH Hönggerberg (HPT), Zürich, Switzerland.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't