Eliminating the isoleucine biosynthetic pathway to reduce competitive carbon outflow during isobutanol production by Saccharomyces cerevisiae.
Eliminating the isoleucine biosynthetic pathway to reduce competitive carbon outflow during isobutanol production by Saccharomyces cerevisiae.
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DOI：
10.1186/s12934-015-0240-6
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发表时间：
2015-04-29
影响因子：
6.4
通讯作者：
Kondo A
中科院分区：
文献类型：
--
作者：
Ida K;Ishii J;Matsuda F;Kondo T;Kondo A
异丁醇是一种重要的生物精炼目标醇，可用作燃料、燃料添加剂或日用化学品。面包酵母（Saccharomyces cerevisiae）由于其耐低pH和耐自溶性，是工业生产异丁醇的有前途的生物。已有报道，直接参与丙酮酸代谢的丙酮酸脱氢酶复合物的基因缺失提高了S.啤酒。然而，可用于S.与细菌宿主如大肠杆菌相比，酿酒酵母的代谢途径是不成熟的，并且除了丙酮酸代谢之外的几种途径与异丁醇生产竞争。删除异丁酸、泛酸或异亮氨酸生物合成途径，以减少与异丁醇生物合成竞争的碳流出。啤酒。明智地消除这些竞争途径增加了异丁醇的生产。ILV 1编码苏氨酸解氨酶，该酶将苏氨酸转化为2-酮丁酸，2-酮丁酸是异亮氨酸生物合成的前体。S. ILV 1缺失的酿酒酵母突变体显示出3.5倍的异丁醇生产率增加。Δ ILV 1策略进一步与两种先前建立的工程化策略（激活埃利希途径的两个步骤和转氢酶样分流）组合，提供了与亲本菌株相比高11倍的异丁醇生产率。该工程菌发酵效价为224 ± 5 mg/L，产量为12.04 ± 0.23 mg/g葡萄糖。删除竞争途径以减少碳流出，包括ILV 1缺失，是增加S.啤酒。
Isobutanol is an important biorefinery target alcohol that can be used as a fuel, fuel additive, or commodity chemical. Baker’s yeast, Saccharomyces cerevisiae, is a promising organism for the industrial manufacture of isobutanol because of its tolerance for low pH and resistance to autolysis. It has been reported that gene deletion of the pyruvate dehydrogenase complex, which is directly involved in pyruvate metabolism, improved isobutanol production by S. cerevisiae. However, the engineering strategies available for S. cerevisiae are immature compared to those available for bacterial hosts such as Escherichia coli, and several pathways in addition to pyruvate metabolism compete with isobutanol production. The isobutyrate, pantothenate or isoleucine biosynthetic pathways were deleted to reduce the outflow of carbon competing with isobutanol biosynthesis in S. cerevisiae. The judicious elimination of these competing pathways increased isobutanol production. ILV1 encodes threonine ammonia-lyase, the enzyme that converts threonine to 2-ketobutanoate, a precursor for isoleucine biosynthesis. S. cerevisiae mutants in which ILV1 had been deleted displayed 3.5-fold increased isobutanol productivity. The ΔILV1 strategy was further combined with two previously established engineering strategies (activation of two steps of the Ehrlich pathway and the transhydrogenase-like shunt), providing 11-fold higher isobutanol productivity as compared to the parent strain. The titer and yield of this engineered strain was 224 ± 5 mg/L and 12.04 ± 0.23 mg/g glucose, respectively. The deletion of competitive pathways to reduce the outflow of carbon, including ILV1 deletion, is an important strategy for increasing isobutanol production by S. cerevisiae.
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影响因子：
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