Department of Pharmaceutical Biology, Groningen Research Institute of Pharmacy, University of Groningen, A, Deusinglaan 1, Groningen 9713AV, The Netherlands.
Microb Cell Fact. 2013 Feb 5;12:13. doi: 10.1186/1475-2859-12-13.
The red yeast Xanthophyllomyces dendrorhous is a natural producer of the carotenoid astaxanthin. Because of its high flux, the native terpene pathway leading to the production of the tetraterpene is of particular interest as it can be redirected toward the production of other terpene compounds. The genetic tools for the transformation of the yeast with the concurrent knock-out of genes involved in the astaxanthin biosynthesis are made available and here we show that the production of the sesquiterpene α-cuprenene is possible in mutant strains of X. dendrorhous transformed with the Cop6 gene originating from the fungus Coprinus cinereus. For the evaluation of the production levels, we chose to express the same gene and analyze the accumulation of α-cuprenene in Escherichia coli and Saccharomyces cerevisiae, as well. Here we propose that X. dendrorhous is a candidate in the search for the potential platform organism for the production of terpenes.
All three X. dendrorhous mutants functionally express the Cop6 gene and accumulate α-cuprenene. The production of α-cuprenene in the red yeast reached 80 mg/L, which represents a far higher concentration compared to the levels obtained in the E. coli and S. cerevisiae mutants. At this expression levels the pool of terpene precursors has not become a limiting factor in the X. dendrorhous mutants since the expression of the Cop6 gene in the genomic rDNA of the yeast allows production of both α-cuprenene and astaxanthin without affecting the growth or the accumulation levels of both compounds.
We have shown that X. dendrorhous can produce α-cuprenene, and the results here presented, next to the capability of accumulating at least two more non-native sesquiterpenes, demonstrates the high potential of this yeast to become an interesting terpene-based drugs producer.
红酵母 Xanthophyllomyces dendrorhous 是虾青素类胡萝卜素的天然生产者。由于其通量高，通向四萜生产的天然萜烯途径特别有趣，因为它可以被重新定向生产其他萜烯化合物。酵母转化的遗传工具与参与虾青素生物合成的基因的敲除同时可用，在这里我们表明，在转化为真菌 Coprinus cinereus 的 Cop6 基因的 X. dendrorhous 突变株中可以生产倍半萜 α-姜烯。为了评估生产水平，我们选择表达相同的基因，并在大肠杆菌和酿酒酵母中分析 α-姜烯的积累。在这里，我们提出 X. dendrorhous 是寻找生产萜烯的潜在平台生物的候选者。
所有三种 X. dendrorhous 突变株均功能性表达 Cop6 基因并积累 α-姜烯。红酵母中 α-姜烯的产量达到 80mg/L，与大肠杆菌和酿酒酵母突变株中获得的水平相比，浓度要高得多。在这种表达水平下，萜烯前体库在 X. dendrorhous 突变株中并未成为限制因素，因为 Cop6 基因在酵母的基因组 rDNA 中的表达允许同时生产 α-姜烯和虾青素，而不会影响两者的生长或积累水平。
我们已经表明 X. dendrorhous 可以生产 α-姜烯，并且这里提出的结果，除了能够积累至少两种其他非天然的倍半萜烯之外，证明了这种酵母具有很高的潜力，可以成为一种有趣的基于萜烯的药物生产菌。
