Linked Life Data

20731719

Source:http://linkedlifedata.com/resource/pubmed/id/20731719

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
19
pubmed:dateCreated
2010-9-15
pubmed:databankReference
pubmed:abstractText
Our previous studies revealed that the double variant of cytochrome P450 (CYP)105A1, R73V/R84A, has a high ability to convert vitamin D(3) to its biologically active form, 1?,25-dihydroxyvitamin D(3) [1?,25(OH)(2)D(3)], suggesting the possibility for R73V/R84A to produce 1?,25(OH)(2)D(3). Because Actinomycetes, including Streptomyces, exhibit properties that have potential advantages in the synthesis of secondary metabolites of industrial and medical importance, we examined the expression of R73V/R84A in Streptomyces lividans TK23 cells under the control of the tipA promoter. As expected, the metabolites 25-hydroxyvitamin D(3) [25(OH)D(3)] and 1?,25(OH)(2)D(3) were detected in the cell culture of the recombinant S. lividans. A large amount of 1?,25(OH)(2)D(3), the second-step metabolite of vitamin D(3), was observed, although a considerable amount of vitamin D(3) still remained in the culture. In addition, novel polar metabolites 1?,25(R),26(OH)(3)D(3) and 1?,25(S),26(OH)(3)D(3), both of which are known to have high antiproliferative activity and low calcemic activity, were observed at a ratio of 5:1. The crystal structure of the double variant with 1?,25(OH)(2)D(3) and a docking model of 1?,25(OH)(2)D(3) in its active site strongly suggest a hydrogen-bond network including the 1?-hydroxyl group, and several water molecules play an important role in the substrate-binding for 26-hydroxylation. In conclusion, we have demonstrated that R73V/R84A can catalyze hydroxylations at C25, C1 and C26 (C27) positions of vitamin D(3) to produce biologically useful compounds.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
1742-4658
pubmed:author
pubmed:copyrightInfo
© 2010 The Authors Journal compilation © 2010 FEBS.
pubmed:issnType
Electronic
pubmed:volume
277
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3999-4009
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
Three-step hydroxylation of vitamin D3 by a genetically engineered CYP105A1: enzymes and catalysis.
pubmed:affiliation
Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, Kurokawa, Imizu, Toyama, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't