Journal
FEBS LETTERS
Volume 588, Issue 18, Pages 3375-3381Publisher
WILEY
DOI: 10.1016/j.febslet.2014.07.028
Keywords
Squalene synthase; Carotenoid synthase; Phytoene desaturase; Directed evolution
Funding
- Japan Society for the Promotion of Science (JSPS) fellowship
- PRESTO (Japan Science and Technology Agency, JST)
- MEXT/JSPS [23108507]
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Squalene synthase (SQS) catalyzes the first step of sterol/hopanoid biosynthesis in various organisms. It has been long recognized that SQSs share a common ancestor with carotenoid synthases, but it is not known how these enzymes selectively produce their own product. In this study, SQSs from yeast, human, and bacteria were independently subjected to directed evolution for the production of the C-30 carotenoid backbone, dehydrosqualene. This was accomplished via high-throughput screening with Pantoea ananatis phytoene desaturase, which can selectively convert dehydrosqualene into yellow carotenoid pigments. Genetic analysis of the resultant mutants revealed various mutations that could effectively convert SQS into a dehydrosqualene synthase.'' All of these mutations are clustered around the residues that have been proposed to be important for NADPH binding. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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