HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Eisenreich, W. Articles by Bacher, A. Search for Related Content PubMed PubMed Citation Articles by Eisenreich, W. Articles by Bacher, A. Social Bookmarking                      What's this?

Volume 272, Number 2, Issue of January 10, 1997 pp. 867-874
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Tracer Studies with Crude U-¹³C-Lipid Mixtures
BIOSYNTHESIS OF THE LIPASE INHIBITOR LIPSTATIN

(Received for publication, July 9, 1996, and in revised form, October 7, 1996)

Wolfgang Eisenreich , Ernst Kupfer ^§ , Wolfgang Weber ^§ and Adelbert Bacher

From the  Department of Organic Chemistry and Biochemistry, Technical University of Munich, Lichtenbergstrasse 4, D-85747 Garching, Federal Republic of Germany and ^§ F. Hoffmann-La Roche Ltd., Pharma Preclinical Research & Development, Department of Biotechnology, CH-4070 Basel, Switzerland

The biosynthesis of the pancreatic lipase inhibitor lipstatin was investigated by fermentation experiments using cultures of Streptomyces toxytricini, which were supplied with soybean oil and a crude mixture of U-¹³C-lipids obtained from algal biomass cultured with ¹³CO₂. Lipstatin was analyzed by one- and two-dimensional NMR spectroscopy. ¹³C total correlation spectroscopy and INADEQUATE experiments show that two fatty acid fragments containing 14 and 8 carbon atoms, respectively, are incorporated en bloc into lipstatin. The 14-carbon fragment is preferentially derived from the unsaturated fatty acid fraction, as shown by an experiment with hydrogenated U-¹³C-lipid mixture, which is conducive to labeling of the 8-carbon moiety but not of the 14-carbon moiety. The data indicate that the lipstatin molecule can be assembled by Claisen condensation of octanoyl-CoA with 3-hydroxy-^5,8-tetradecanoyl-CoA obtained by  oxidation of linoleic acid. The formation of lipstatin from acetate units by a polyketide-type pathway is ruled out conclusively by these data. The data show that surprisingly clear labeling patterns can be obtained in studies with crude, universally ¹³C-labeled precursor mixtures that are proffered together with a large excess of unlabeled material. One- and two-dimensional ¹³C total correlation spectroscopy analyses are suggested as elegant methods for the delineation of contiguously ¹³C-labeled biosynthetic blocks.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				I. Werner, A. Bacher, and W. Eisenreich Retrobiosynthetic NMR Studies with 13C-Labeled Glucose. FORMATION OF GALLIC ACID IN PLANTS AND FUNGI J. Biol. Chem., October 10, 1997; 272(41): 25474 - 25482. [Abstract] [Full Text] [PDF]

				M. Goese, W. Eisenreich, E. Kupfer, W. Weber, and A. Bacher Biosynthetic Origin of Hydrogen Atoms in the Lipase Inhibitor Lipstatin J. Biol. Chem., July 7, 2000; 275(28): 21192 - 21196. [Abstract] [Full Text] [PDF]