Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, ul. Wojska Polskiego 48, 60-627 Poznan, Poland.
Imperial College Centre for Synthetic Biology and Department of Bioengineering, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
FEMS Yeast Res. 2019 Mar 1;19(2). doi: 10.1093/femsyr/foy122.
Microbial cells can produce a vast spectrum of chemical compounds, including those most desired by the global chemical market, for example, higher alcohols, which are promising alternative fuels and chemical feedstock. In the current research, we investigated the effects of the Ehrlich pathway genetic engineering on higher alcohols production in Yarrowialipolytica, which directly follows our previous findings concerning elucidation of putative molecular identities involved in this pathway. To this end, we constructed two alternative expression cassettes composed of previously identified genes, putatively involved in the Ehrlich pathway in Y. lipolytica, and cloned them under the control of constitutive pTEF promoter, and by this released them from extensive native regulation. The effects of the pathway engineering were investigated upon provision of different Ehrlich pathway-inducing amino acids (L-Phe, L-Leu, L-Ile and L-Val). In general, amplification of the Ehrlich pathway in many cases led to increased formation of a respective higher alcohol from its precursor. We observed interesting effects of aminotransferase BAT2 deletion on synthesis of 2-phenylethanol and its acetate ester, significant relationship between L-Val and L-Phe catabolic pathways and extensive 'cross-induction' of the derivative compounds synthesis by non-direct precursors.
微生物细胞可以产生广泛的化合物，包括全球化学市场最需要的化合物，例如，高级醇，这是很有前途的替代燃料和化学原料。在目前的研究中，我们研究了 Ehrlich 途径遗传工程对解脂耶氏酵母中高级醇生产的影响，这直接遵循了我们之前关于阐明该途径中涉及的假定分子身份的发现。为此，我们构建了两个替代的表达盒，由先前鉴定的、可能参与 Y. lipolytica Ehrlich 途径的基因组成，并在组成型 pTEF 启动子的控制下克隆它们，从而摆脱了广泛的天然调控。通过提供不同的 Ehrlich 途径诱导氨基酸（L-Phe、L-Leu、L-Ile 和 L-Val）来研究途径工程的影响。一般来说，Ehrlich 途径的扩增在许多情况下会导致相应的高级醇从其前体形成增加。我们观察到 BAT2 转氨酶缺失对 2-苯乙醇及其醋酸酯合成的有趣影响，L-Val 和 L-Phe 分解代谢途径之间的显著关系以及非直接前体对衍生化合物合成的广泛“交叉诱导”。
