Method for production of isoprenoid compounds

摘要

The present invention is directed to variant squalene synthase enzymes, including Saccharomyces cerevisiae squalene synthase enzymes, and to nucleic acid molecules encoding these variant enzymes. These variant enzymes produce squalene at a lower rate than the wild-type enzyme, allowing more farnesyl pyrophosphate to be utilized for production of isoprenoid compounds, while still producing sufficient squalene to allow the S. cerevisiae cells to grow without the requirement for supplementation by sterols such as ergosterol. These variant enzymes, therefore, are highly suitable for the efficient production of isoprenoids.

主权项

1. A nucleic acid molecule that encodes a variant squalene synthase enzyme, wherein the variant enzyme comprises a sequence of amino acid residues that is at least 95% identical to the squalene synthase enzyme whose sequence is set forth in SEQ ID NO: 4, whereby:
the variant enzyme exhibits reduced cellular squalene synthase activity compared to the enzyme whose sequence is set forth in SEQ ID NO: 4; the variant enzyme, when present and expressed in vivo in a eukaryotic microbial host as the only squalene synthase species, catalyzes the synthesis of squalene at a sufficiently high rate such that supplementation of the eukaryotic microbial host with a sterol is not required for growth; and the eukaryotic microbial host produces a terpene product in medium lacking a sterol.