Production of α-cuprenene in Xanthophyllomyces dendrorhous: a step closer to a potent terpene biofactory.
Production of α-cuprenene in Xanthophyllomyces dendrorhous: a step closer to a potent terpene biofactory.
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DOI：
10.1186/1475-2859-12-13
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发表时间：
2013-02-05
影响因子：
6.4
通讯作者：
Kayser O
中科院分区：
文献类型：
--
作者：
Melillo E;Setroikromo R;Quax WJ;Kayser O
红酵母Xanthophyllomyces dendrorhous是类胡萝卜素虾青素的天然生产者。由于其高通量，导致产生四萜的天然萜途径特别令人感兴趣，因为它可以被重新导向其它萜化合物的产生。用于转化酵母并同时敲除参与虾青素生物合成的基因的遗传工具是可用的，并且在这里我们表明在X的突变株中生产倍半萜α-cuprenene是可能的。用来源于真菌灰盖鬼伞的Cop 6基因转化的树枝状。为了评估生产水平，我们选择了在大肠杆菌和酿酒酵母中表达相同的基因，并分析了α-cuprenene的积累。这里我们提出X.在寻找用于生产萜类的潜在平台生物体中，Dendornhous是一种候选物。全部三个X。树枝状突变体在功能上表达Cop 6基因并积累α-蒎烯。在此条件下，α-蒎烯在红酵母中的产量达到80 mg/L，远远高于E. coli和革兰氏阳性菌S.酿酒酵母突变体。在该表达水平下，萜前体库没有成为X.因为Cop 6基因在酵母的基因组rDNA中的表达允许α-杯异戊二烯和虾青素的产生，而不影响这两种化合物的生长或积累水平。我们已经证明了X。树枝状酵母可以产生α-蒎烯，本文所述的结果，除了能够积累至少两种以上的非天然倍半萜外，还证明了该酵母成为令人感兴趣的萜类药物生产者的高潜力。
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.
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DOI：
10.1126/science.1191652
发表时间：
2010-10-01
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Ajikumar PK;Xiao WH;Tyo KE;Wang Y;Simeon F;Leonard E;Mucha O;Phon TH;Pfeifer B;Stephanopoulos G
通讯作者：
Stephanopoulos G
影响因子：
4.4
作者：
Linger, Jeffrey G.;Adney, William S.;Darzins, Al
通讯作者：
Darzins, Al
DOI：
10.1073/pnas.1110740109
发表时间：
2012-01-17
影响因子：
11.1
作者：
Westfall, Patrick J.;Pitera, Douglas J.;Paddon, Chris J.
通讯作者：
Paddon, Chris J.
影响因子：
6.7
作者：
NIKLITSCHEK, MAURICIO;ALCAÍNO, JENNIFER;CIFUENTES, VÍCTOR
通讯作者：
CIFUENTES, VÍCTOR
影响因子：
3.8
作者：
Newman, Jack D.;Marshall, Jessica;Keasling, Jay D.
通讯作者：
Keasling, Jay D.