Improving yeast strains using recyclable integration cassettes, for the production of plant terpenoids

Microbial Cell Factories

Research

Sotirios C Kampranis¹  Ivana Cvetkovic¹  Christopher B Johnson¹  Codruta Ignea²  Sofia Loupassaki³  Panagiotis Kefalas³  Antonios M Makris⁴ 

[1] Department of Natural Products and Biotechnology, Centre International de Hautes Etudes Agronomiques Méditerranéennes. Mediterranean Agronomic Institute of Chania, P.O. Box 85, 73100, Chania, Greece;Department of Natural Products and Biotechnology, Centre International de Hautes Etudes Agronomiques Méditerranéennes. Mediterranean Agronomic Institute of Chania, P.O. Box 85, 73100, Chania, Greece;Department of Biology, University of Crete, Vasilika Vouton, Heraklio, 71409, Crete, Greece;Department of Natural Products and Biotechnology, Centre International de Hautes Etudes Agronomiques Méditerranéennes. Mediterranean Agronomic Institute of Chania, P.O. Box 85, 73100, Chania, Greece;Department of Food Quality and Chemistry of Natural Products, Centre International de Hautes Etudes Agronomiques Méditerranéennes. Mediterranean Agronomic Institute of Chania, P.O. Box 85, 73100, Chania, Greece;Department of Natural Products and Biotechnology, Centre International de Hautes Etudes Agronomiques Méditerranéennes. Mediterranean Agronomic Institute of Chania, P.O. Box 85, 73100, Chania, Greece;Institute of Agrobiotechnology/CERTH, P.O.Box 60361, Thermi, 57001, Thessaloniki, Greece;

关键词: SPME;    Terpenoid;    Monoterpene;    Sesquiterpene;    Cineole;   

DOI  :  10.1186/1475-2859-10-4

 received in 2010-08-03, accepted in 2011-01-28,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundTerpenoids constitute a large family of natural products, attracting commercial interest for a variety of uses as flavours, fragrances, drugs and alternative fuels. Saccharomyces cerevisiae offers a versatile cell factory, as the precursors of terpenoid biosynthesis are naturally synthesized by the sterol biosynthetic pathway.ResultsS. cerevisiae wild type yeast cells, selected for their capacity to produce high sterol levels were targeted for improvement aiming to increase production. Recyclable integration cassettes were developed which enable the unlimited sequential integration of desirable genetic elements (promoters, genes, termination sequence) at any desired locus in the yeast genome. The approach was applied on the yeast sterol biosynthetic pathway genes HMG2, ERG20 and IDI1 resulting in several-fold increase in plant monoterpene and sesquiterpene production. The improved strains were robust and could sustain high terpenoid production levels for an extended period. Simultaneous plasmid-driven co-expression of IDI1 and the HMG2 (K6R) variant, in the improved strain background, maximized monoterpene production levels. Expression of two terpene synthase enzymes from the sage species Salvia fruticosa and S. pomifera (SfCinS1, SpP330) in the modified yeast cells identified a range of terpenoids which are also present in the plant essential oils. Co-expression of the putative interacting protein HSP90 with cineole synthase 1 (SfCinS1) also improved production levels, pointing to an additional means to improve production.ConclusionsUsing the developed molecular tools, new yeast strains were generated with increased capacity to produce plant terpenoids. The approach taken and the durability of the strains allow successive rounds of improvement to maximize yields.

【 授权许可】

Unknown   
© Ignea et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.