Jin V Lee1, Ran Furman1, Paul H Axelsen2. 1. Department of Pharmacology, University of Pennsylvania, Philadelphia, PA 19104, United States. 2. Department of Pharmacology, University of Pennsylvania, Philadelphia, PA 19104, United States; Departments of Biochemistry and Biophysics, and Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States. Electronic address: axe@pharm.med.upenn.edu.
Abstract
Arachidonic acid (ARA) is one of the most abundant polyunsaturated fatty acids (PUFAs) in the mammalian brain. Many enzymatically- and nonenzymatically-produced metabolic products have important and potent pharmacological properties. However, uniformly isotope labeled forms of ARA are not commercially available for studying the metabolic fates of ARA. This study describes a simple and efficient protocol for the biosynthesis of U-13C-ARA from U-13C-glucose, and U-14C-ARA from U-14C-glucose by Mortierella alpina. The protocols yield approximately 100nmol quantities of U-13C-ARA with an isotopic purity of 95% from a 500μl batch volume, and approximately 2μCi quantities of U-14C-ARA with an apparent specific activity in excess of 1200Ci/mol from a 250μl batch volume.
Arachidonic acid (n class="Chemical">ARA) is one of the most abundant polyunsaturated fatty acids (PUFAs) in the mammalian brain. Many enzymatically- and nonenzymatically-produced metabolic products have important and potent pharmacological properties. However, uniformly isotope labeled forms of ARA are not commercially available for studying the metabolic fates of ARA. This study describes a simple and efficient protocol for the biosynthesis of U-13C-ARA from U-13C-glucose, and U-14C-ARA from U-14C-glucose by Mortierella alpina. The protocols yield approximately 100nmol quantities of U-13C-ARA with an isotopic purity of 95% from a 500μl batch volume, and approximately 2μCi quantities of U-14C-ARA with an apparent specific activity in excess of 1200Ci/mol from a 250μl batch volume.
Authors: J D Morrow; T A Minton; C R Mukundan; M D Campbell; W E Zackert; V C Daniel; K F Badr; I A Blair; L J Roberts Journal: J Biol Chem Date: 1994-02-11 Impact factor: 5.157