Enhancement of astaxanthin production in Xanthophyllomyces dendrorhous by efficient method for the complete deletion of genes.
Enhancement of astaxanthin production in Xanthophyllomyces dendrorhous by efficient method for the complete deletion of genes.
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DOI：
10.1186/s12934-016-0556-x
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发表时间：
2016-09-13
影响因子：
6.4
通讯作者：
Kondo A
中科院分区：
文献类型：
--
作者：
Yamamoto K;Hara KY;Morita T;Nishimura A;Sasaki D;Ishii J;Ogino C;Kizaki N;Kondo A
红酵母，Xanthophyllomyces dendronorhous是已知唯一能产生虾青素的酵母，虾青素是一种抗氧化剂类异戊二烯（类胡萝卜素），广泛应用于水产养殖、食品、制药和化妆品行业。这种微生物作为类异戊二烯生产的平台细胞工厂的潜力已经被认识到，因为通过其天然萜烯途径的高通量。最近，我们在X.树状体和增强甲羟戊酸合成途径，以增加虾青素的生产。相反，甲羟戊酸合成途径通过反馈抑制被麦角固醇抑制。因此，通过删除参与麦角固醇合成的基因来释放甲羟戊酸合成途径的这种抑制是改善类异戊二烯生产的有前景的策略。提出了一种有效的删除二倍体基因的方法。然而，尚未开发出树状结构。在引入抗生素抗性基因以用靶基因替换后，在逐渐增加的抗生素浓度下培养Xanthophyllomyces dendrohous。用这种方法，双CYP61基因编码的C-22甾醇脱氢酶有关麦角甾醇生物合成的顺序删除。与亲本菌株和单CYP 61破坏菌株相比，该双CYP 61缺失菌株显示降低的麦角固醇生物合成。此外，与亲本菌株和单一CYP 61敲除菌株相比，CYP 61基因的这种双缺失显示出增加的虾青素产量。最后，与亲本菌株相比，虾青素产量提高了1.4倍，尽管在单一CYP 61敲除菌株中虾青素产量不受影响。在本研究中，我们开发了一个系统，完全删除目的二倍体基因在X。树枝状的使用这种方法，我们删除了参与麦角固醇合成的二倍体CYP61基因，麦角固醇抑制甲羟戊酸途径，甲羟戊酸是类异戊二烯生物合成的常见底物。麦角固醇生物合成的减少增加了虾青素的产生。本研究所开发的二倍体基因缺失方法具有提高类异戊二烯类化合物工业生产的潜力。树枝状的本文的在线版本（doi：10.1186/s12934 - 016 - 0556-x）包含补充材料，可供授权用户使用。
Red yeast, Xanthophyllomyces dendrorhous is the only yeast known to produce astaxanthin, an anti-oxidant isoprenoid (carotenoid) widely used in the aquaculture, food, pharmaceutical and cosmetic industries. The potential of this microorganism as a platform cell factory for isoprenoid production has been recognized because of high flux through its native terpene pathway. Recently, we developed a multiple gene expression system in X. dendrorhous and enhanced the mevalonate synthetic pathway to increase astaxanthin production. In contrast, the mevalonate synthetic pathway is suppressed by ergosterol through feedback inhibition. Therefore, releasing the mevalonate synthetic pathway from this inhibition through the deletion of genes involved in ergosterol synthesis is a promising strategy to improve isoprenoid production. An efficient method for deleting diploid genes in X. dendrorhous, however, has not yet been developed. Xanthophyllomyces dendrorhous was cultivated under gradually increasing concentrations of antibiotics following the introduction of antibiotic resistant genes to be replaced with target genes. Using this method, double CYP61 genes encoding C-22 sterol desaturases relating to ergosterol biosynthesis were deleted sequentially. This double CYP61 deleted strain showed decreased ergosterol biosynthesis compared with the parental strain and single CYP61 disrupted strain. Additionally, this double deletion of CYP61 genes showed increased astaxanthin production compared with the parental strain and the single CYP61 knockout strain. Finally, astaxanthin production was enhanced by 1.4-fold compared with the parental strain, although astaxanthin production was not affected in the single CYP61 knockout strain. In this study, we developed a system to completely delete target diploid genes in X. dendrorhous. Using this method, we deleted diploid CYP61 genes involved in the synthesis of ergosterol that inhibits the pathway for mevalonate, which is a common substrate for isoprenoid biosynthesis. The resulting decrease in ergosterol biosynthesis increased astaxanthin production. The efficient method for deleting diploid genes developed in this study has the potential to improve industrial production of various isoprenoids in X. dendrorhous. The online version of this article (doi:10.1186/s12934-016-0556-x) contains supplementary material, which is available to authorized users.
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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
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：
5
作者：
Hara, Kiyotaka Y.;Morita, Toshihiko;Kondo, Akihiko
通讯作者：
Kondo, Akihiko
影响因子：
3.5
作者：
Wery, J;Gutker, D;vanOoyen, AJJ
通讯作者：
vanOoyen, AJJ
影响因子：
6.4
作者：
Melillo E;Setroikromo R;Quax WJ;Kayser O
通讯作者：
Kayser O