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. 2015 Apr 24;14:60. doi: 10.1186/s12934-015-0246-0

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© Trikka et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Maximizing sclareol titers in AM238 cells. (A) EG60 and AM238 cells expressing CLS-ERG20 (F96C) only, CLS-ERG20 (F96C) with sclareol synthase SCLS, and CLS-ERG20 (F96C), SCLS with CD-HMG2. The engineered iterative perturbations generated a strain producing 12-fold higher sclareol levels than wild type cells; (B) Released NOH + FOH and GLOH + GGOH in EG60 and AM238 cells expressing the three plasmids. A reduction of released terpenols is seen in AM238 cells indicating a more efficient substrate conversion to sclareol; (C) Ergosterol and squalene levels in EG60 and AM238 cells expressing the three plasmids. Ergosterol levels are reduced by half in AM238 cells co-expressing SCLS and CD-HMG2, compared to CLS-ERG20 (F96C) only. Squalene levels are significantly increased by CD-HMG2 expression.