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A new method for isolating physiologically active Mg-protoporphyrin

Gough, Simon LU ; Rzeznicka, Kamila LU ; Peterson Wulff, Ragna LU ; da Cruz Francisco, José LU ; Hansson, Andreas LU ; Jensen, Poul Erik and Hansson, Mats LU (2007) In Plant Physiology and Biochemistry 45(12). p.932-936
Abstract
Mg-protoporphyrin monomethyl ester (MPE) is a biosynthetic intermediate of chlorophyll and converted by MPE cyclase to protochlorophyllide.

Limited availability of MPE has so far hampered cyclase research. In a new, simplified, method MPE was prepared from freeze dried bchE

mutant Rhodobacter capsulatus DB575 cells by extraction with acetone/H2O/25% NH3. Isolated MPE was identified by absorption and fluorescence

spectroscopy, and its purity was analyzed by HPLC. The extracted MPE was dried and redissolved in buffered DMSO and its substrate

activity is shown by enzymatic cyclase assays. A linear time course was observed for MPE conversion to protochlorophyllide by enzymes from

barley etioplasts.... (More)
Mg-protoporphyrin monomethyl ester (MPE) is a biosynthetic intermediate of chlorophyll and converted by MPE cyclase to protochlorophyllide.

Limited availability of MPE has so far hampered cyclase research. In a new, simplified, method MPE was prepared from freeze dried bchE

mutant Rhodobacter capsulatus DB575 cells by extraction with acetone/H2O/25% NH3. Isolated MPE was identified by absorption and fluorescence

spectroscopy, and its purity was analyzed by HPLC. The extracted MPE was dried and redissolved in buffered DMSO and its substrate

activity is shown by enzymatic cyclase assays. A linear time course was observed for MPE conversion to protochlorophyllide by enzymes from

barley etioplasts. Our innovation of freeze drying the R. capsulatus cells before extraction provides a high yield method for MPE, which is significantly

faster and more reproducible than previous extraction methods. (Less)
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author
organization
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type
Contribution to journal
publication status
published
subject
keywords
Mg-protoporphyrin monomethyl ester, MPE, Hordeum vulgare, Cyclase, Rhodobacter capsulatus, Chlorophyll biosynthesis
in
Plant Physiology and Biochemistry
volume
45
issue
12
pages
932 - 936
publisher
Elsevier
external identifiers
  • wos:000252077500009
  • scopus:36248975616
ISSN
1873-2690
DOI
10.1016/j.plaphy.2007.09.001
language
English
LU publication?
yes
id
e02f7c61-6316-4ed0-9f5d-bd37fa0beea5 (old id 935809)
date added to LUP
2008-01-24 17:59:24
date last changed
2017-01-01 07:20:29
@article{e02f7c61-6316-4ed0-9f5d-bd37fa0beea5,
  abstract     = {Mg-protoporphyrin monomethyl ester (MPE) is a biosynthetic intermediate of chlorophyll and converted by MPE cyclase to protochlorophyllide.<br/><br>
Limited availability of MPE has so far hampered cyclase research. In a new, simplified, method MPE was prepared from freeze dried bchE<br/><br>
mutant Rhodobacter capsulatus DB575 cells by extraction with acetone/H2O/25% NH3. Isolated MPE was identified by absorption and fluorescence<br/><br>
spectroscopy, and its purity was analyzed by HPLC. The extracted MPE was dried and redissolved in buffered DMSO and its substrate<br/><br>
activity is shown by enzymatic cyclase assays. A linear time course was observed for MPE conversion to protochlorophyllide by enzymes from<br/><br>
barley etioplasts. Our innovation of freeze drying the R. capsulatus cells before extraction provides a high yield method for MPE, which is significantly<br/><br>
faster and more reproducible than previous extraction methods.},
  author       = {Gough, Simon and Rzeznicka, Kamila and Peterson Wulff, Ragna and da Cruz Francisco, José and Hansson, Andreas and Jensen, Poul Erik and Hansson, Mats},
  issn         = {1873-2690},
  keyword      = {Mg-protoporphyrin monomethyl ester,MPE,Hordeum vulgare,Cyclase,Rhodobacter capsulatus,Chlorophyll biosynthesis},
  language     = {eng},
  number       = {12},
  pages        = {932--936},
  publisher    = {Elsevier},
  series       = {Plant Physiology and Biochemistry},
  title        = {A new method for isolating physiologically active Mg-protoporphyrin},
  url          = {http://dx.doi.org/10.1016/j.plaphy.2007.09.001},
  volume       = {45},
  year         = {2007},
}