Positive genetic interactors of HMG2 identify a new set of genetic perturbations for improving sesquiterpene production in Saccharomyces cerevisiae.
Positive genetic interactors of HMG2 identify a new set of genetic perturbations for improving sesquiterpene production in Saccharomyces cerevisiae.
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DOI：
10.1186/1475-2859-11-162
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发表时间：
2012-12-22
影响因子：
6.4
通讯作者：
Makris AM
中科院分区：
文献类型：
--
作者：
Ignea C;Trikka FA;Kourtzelis I;Argiriou A;Kanellis AK;Kampranis SC;Makris AM
萜类和类异戊二烯是一类重要的天然产物，包括目前使用的药物、高价值的生物活性化合物和工业化合物以及燃料候选物。由于它们的工业应用，S.酿酒酵母菌株能够产生高水平的萜类化合物。为了鉴定可被操纵以增加倍半萜产生的新基因靶标，在存在或不存在稳定的HMG 2（K6 R）过表达的情况下，将一组HMG 2阳性遗传相互作用物评估为单基因和双基因杂合缺失。在单个等位基因缺失后，检测的大多数基因导致酵母细胞中倍半萜烯产量增加。一组三个基因的串联杂合缺失，泛素连接酶ubc 7和ssm 4/doa 10，和ER常驻蛋白pho 86，导致11倍增加的卡隆产量（125毫克/升，摇瓶）相比，缺乏这些修改的细胞。杂合缺失的影响似乎是由于Hmg 1 p和Hmg 2 p稳定。杂合缺失导致蛋白质水平显著降低，但不会导致在单倍体菌株中常见的生长障碍。通过利用酵母中理想的单倍不稳定性，我们鉴定了一组新的基因，其可以串联破坏并引起Hmgp的显著稳定和倍半萜生产的大幅增加。这里提出的方法允许在酵母细胞工厂菌株上编译新的遗传扰动，而不会对细胞生长和活力产生负面影响。
Terpenoids and isoprenoids are an important class of natural products, which includes currently used drugs, high value bioactive and industrial compounds, and fuel candidates. Due to their industrial application, there is increasing interest in the development of S. cerevisiae strains capable of producing high levels of terpenoids. Aiming to identify new gene targets which can be manipulated to increase sesquiterpene production, a set of HMG2 positive genetic interactors were assessed as single and digenic heterozygous deletions in the presence or absence of stable HMG2(K6R) overexpression. Upon single allele deletion, most genes examined led to increased sesquiterpene production in yeast cells. Tandem heterozygous deletion of a set of three genes, the ubiquitin ligases ubc7 and ssm4/doa10, and the ER resident protein pho86, led to an 11-fold increase in caryophyllene yields (125 mg/L in shake flasks) compared to cells lacking these modifications. The effect of the heterozygous deletions appears to be due to Hmg1p and Hmg2p stabilization. Heterozygous deletions cause significant reductions in protein levels but do not lead to growth impediments frequently seen in haploid strains. By exploiting desirable haploinsufficiencies in yeast, we identified a new set of genes that can be disrupted in tandem and cause significant stabilization of Hmgp and a substantial increase in sesquiterpene production. The approach presented here allows new genetic perturbations to be compiled on yeast cell factory strains without negatively impacting cell growth and viability.
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影响因子：
9.9
作者：
通讯作者：
--
影响因子：
8.4
作者：
Asadollahi, Mohammad A.;Maury, Jerome;Nielsen, Jens
通讯作者：
Nielsen, Jens
影响因子：
3.3
作者：
Federovitch, Christine M.;Jones, Ying Z.;Hampton, Randolph Y.
通讯作者：
Hampton, Randolph Y.
影响因子：
4
作者：
Leadsham, Jane E.;Miller, Katherine;Gourlay, Campbell W.
通讯作者：
Gourlay, Campbell W.
DOI：
10.1126/science.1180823
发表时间：
2010-01-22
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Costanzo M;Baryshnikova A;Bellay J;Kim Y;Spear ED;Sevier CS;Ding H;Koh JL;Toufighi K;Mostafavi S;Prinz J;St Onge RP;VanderSluis B;Makhnevych T;Vizeacoumar FJ;Alizadeh S;Bahr S;Brost RL;Chen Y;Cokol M;Deshpande R;Li Z;Lin ZY;Liang W;Marback M;Paw J;San Luis BJ;Shuteriqi E;Tong AH;van Dyk N;Wallace IM;Whitney JA;Weirauch MT;Zhong G;Zhu H;Houry WA;Brudno M;Ragibizadeh S;Papp B;Pál C;Roth FP;Giaever G;Nislow C;Troyanskaya OG;Bussey H;Bader GD;Gingras AC;Morris QD;Kim PM;Kaiser CA;Myers CL;Andrews BJ;Boone C
通讯作者：
Boone C