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首页> 外文期刊>Journal of Biotechnology >Artificial microRNA-mediated knockdown of pyruvate formate lyase (PFL1) provides evidence for an active 3-hydroxybutyrate production pathway in the green alga Chlamydomonas reinhardtii
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Artificial microRNA-mediated knockdown of pyruvate formate lyase (PFL1) provides evidence for an active 3-hydroxybutyrate production pathway in the green alga Chlamydomonas reinhardtii

机译：人工microRNA介导的丙酮酸甲酸裂解酶（PFL1）的敲除为绿藻衣藻（Chlamydomonas reinhardtii）产生活跃的3-羟基丁酸生产途径提供了证据

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摘要

Artificial microRNA technology was investigated as a means of down regulating metabolic pathways in the green alga Chlamydomonas reinhardtii, targeting pyruvate formate lyase (PFL1), which catalyzes the conversion of pyruvate to acetyl-CoA and formate during anoxic conditions. Two transformants with an 80–90% reduction in target protein and mRNA levels were identified. Nuclear magnetic resonance spectroscopy confirmed a substantial decrease in the production of formate in the knockdown lines during dark anoxic conditions and a re-routing of metabolism leading to enhanced production of ethanol and lactate. Under microaerobic conditions in the light, induced by sulphur-deprivation, knock-down of PFL1 resulted in reduced formate and ethanol production, increased net consumption of acetate and the excretion of lactate but no increase in the production of hydrogen. In addition the production of 3-hydroxybutyrate was identified in knock-down line cultures during the transition between microaerobic and anoxic conditions. Overall our results indicate that microRNA knock-down is a useful tool to manipulate anaerobic metabolism in C. reinhardtii.
机译：研究了人工microRNA技术，作为下调莱茵藻衣藻中代谢途径的一种手段，其靶向丙酮酸甲酸裂解酶（PFL1），在缺氧条件下催化丙酮酸转化为乙酰辅酶A和甲酸。确定了两个目标蛋白和mRNA水平降低80-90％的转化体。核磁共振波谱证实，在黑暗缺氧条件下，击倒管线中甲酸盐的产生显着减少，并且代谢重新路由，导致乙醇和乳酸的产生增加。在光的微需氧条件下，由于硫被剥夺，PFL1的敲低导致甲酸和乙醇的产生减少，乙酸盐的净消耗增加和乳酸的排泄，但氢的产生没有增加。另外，在微需氧条件和缺氧条件之间的过渡过程中，在组合生产线培养物中鉴定出3-羟基丁酸酯的产生。总的来说，我们的结果表明，microRNA敲低是操纵莱茵衣藻厌氧代谢的有用工具。

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