Linked Life Data

16575563

Source:http://linkedlifedata.com/resource/pubmed/id/16575563

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
2006-10-12
pubmed:abstractText
Spirulina-acyl-lipid desaturases are integral membrane proteins found in thylakoid and plasma membranes. These enzymes catalyze the fatty acid desaturation process of Spirulina to yield gamma-linolenic acid (GLA) as the final desaturation product. It has been reported that the cyanobacterial desaturases use ferredoxin as an electron donor, whereas the acyl-lipid desaturase in plant cytoplasm and the acyl-CoA desaturase of animals and fungi use cytochrome b (5). The low level of ferredoxin present in Escherichia coli cells leads to an inability to synthesize GLA when the cells are transformed with the Spirulina-(6) desaturase, desD, and grown in the presence of the reaction substrate, linoleic acid. In this study, Spirulina-(6) desaturase, encoded by the desD gene, was N-terminally fused and co-expressed with the cytochrome b (5) domain from Mucor rouxii. The product, GLA, made heterologously in E. coli and Saccharomyces cerevisiae, was then detected and analyzed. The results revealed the production of GLA by Spirulina-(6) desaturase fused or co-expressed with cytochrome b (5) in E. coli cells, in which GLA production by this gene cannot occur in the absence of cytochrome b (5). Moreover, the GLA production ability in the E. coli host cells was lost after the single substitution mutation was introduced to H52 in the HPGG motif of the cytochrome b (5) domain. These results revealed the complementation of the ferredoxin requirement by the fusion or co-expression of the fungal-cytochrome b (5) domain in the desaturation process of Spirulina-(6) desaturase. Furthermore, the free form of cytochrome b (5) domain can also enhance GLA production by the Spirulina-desD gene in yeast cells.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0175-7598
pubmed:author
pubmed:issnType
Print
pubmed:volume
72
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1192-201
pubmed:meshHeading
pubmed-meshheading:16575563-Amino Acid Motifs, pubmed-meshheading:16575563-Amino Acid Sequence, pubmed-meshheading:16575563-Amino Acid Substitution, pubmed-meshheading:16575563-Artificial Gene Fusion, pubmed-meshheading:16575563-Bacterial Proteins, pubmed-meshheading:16575563-Blotting, Western, pubmed-meshheading:16575563-Cytochromes b5, pubmed-meshheading:16575563-Escherichia coli, pubmed-meshheading:16575563-Ferredoxins, pubmed-meshheading:16575563-Fungal Proteins, pubmed-meshheading:16575563-Gas Chromatography-Mass Spectrometry, pubmed-meshheading:16575563-Gene Expression, pubmed-meshheading:16575563-Linoleoyl-CoA Desaturase, pubmed-meshheading:16575563-Molecular Sequence Data, pubmed-meshheading:16575563-Mucor, pubmed-meshheading:16575563-Mutagenesis, Site-Directed, pubmed-meshheading:16575563-Mutation, pubmed-meshheading:16575563-Recombinant Fusion Proteins, pubmed-meshheading:16575563-Saccharomyces cerevisiae, pubmed-meshheading:16575563-Spirulina, pubmed-meshheading:16575563-gamma-Linolenic Acid
pubmed:year
2006
pubmed:articleTitle
Revealing the complementation of ferredoxin by cytochrome b (5) in the Spirulina- (6)-desaturation reaction by N-terminal fusion and co-expression of the fungal-cytochrome b (5) domain and Spirulina- (6)-acyl-lipid desaturase.
pubmed:affiliation
National Center for Genetic Engineering and Biotechnology, 83 Moo8, Thakham, Bangkhuntien, Bangkok, 10150, Thailand. apiradee@biotec.or.th
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't