WILLIBALD, B and BOVES, H and HOLLER, E (1995) PREPARATIVE SYNTHESIS OF BETA-L-MALYL-COENZYME-A ASSISTED BY MALYL-COENZYME-A SYNTHETASE FROM PSEUDOMONAS AM1. ANALYTICAL BIOCHEMISTRY, 227 (2). pp. 363-367. ISSN 0003-2697,
Full text not available from this repository.Abstract
beta-L-Malyl-CoA was synthesized from L-malate, CoA, and ATP in the presence of catalytic amounts of L-malyl-CoA synthetase (thiokinase) from Pseudomona AM1, which had been 50-fold purified by protamine sulfate precipitation, ammonium sulfate precipitation, chromatography on DEAE-cellulose, and affinity chromatography on High Trap Blue in less than 2 days. The homogeneous enzyme was free of L-malyl-CoA lyase and showed 63% homology with succinyl-CoA synthetase from Thermus aquaticus in its N-terminal sequences. Yields of beta-L-[C-14]malyl-CoA (1-10 mu mol) were 70% before and 65% after purification in 0.1-0.5 mu mol portions by high-performance liquid chromatography on a MN Nucleosil 100-7 C8 column. For most biochemical work, the product was partially purified with an overall 45% yield by chromatography on DEAE-Sephacel. The identity of the compound as beta-L-malyl-CoA was verified by chemical and enzymatic tests, and also in comparison with its chemically synthesized counterpart. The enzymatic synthesis, especially of radioactively labeled beta-L-malyl-CoA, is considerably faster, higher in yield, and less problematic than chemical synthesis. (C) 1995 Academic Press, Inc.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CLEAVAGE; PROTEINS; GELS; |
| Depositing User: | Dr. Gernot Deinzer |
| Last Modified: | 19 Oct 2022 08:37 |
| URI: | https://pred.uni-regensburg.de/id/eprint/52533 |
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