期刊
SYNTHETIC AND SYSTEMS BIOTECHNOLOGY
卷 7, 期 1, 页码 498-505出版社
KEAI PUBLISHING LTD
DOI: 10.1016/j.synbio.2021.12.005
关键词
Ogataea polymorpha; Promoter; Hybrid promoter; Upstream activation sequence; Metabolic engineering; Fatty alcohols
资金
- National Natural Science Foundation of China [21808216, 22161142008, M-0246]
- Key project at central government level: The ability establishment of sustainable use for valuable Chinese medicine resources [2060302]
- DICP innovation grant from Dalian Institute of Chemicals Physics, CAS [DICP I202021, I201920]
This study systematically characterized native promoters in Ogataea polymorpha and obtained a group of constitutive, inducible, and hybrid promoters, which enriched the promoter toolbox for metabolic engineering of O. polymorpha and provided an important resource for its bio-manufacturing.
Bio-manufacturing via microbial cell factory requires large promoter library for fine-tuned metabolic engineering. Ogataea polymorpha, one of the methylotrophic yeasts, possesses advantages in broad substrate spectrum, thermal-tolerance, and capacity to achieve high-density fermentation. However, a limited number of available promoters hinders the engineering of O. polymorpha for bio-productions. Here, we systematically characterized native promoters in O. polymorpha by both GFP fluorescence and fatty alcohol biosynthesis. Ten constitutive promoters (P-PDH, P-PYK, P-FBA, P-PGM, P-GLK, P-TRI, P-GPI, P-ADH1, P-TEF1 and P-GCW14) were obtained with the activity range of 13%-130% of the common promoter P-GAP (the promoter of glyceraldehyde-3-phosphate dehydrogenase), among which P-PDH and P-GCW14 were further verified by biosynthesis of fatty alcohol. Furthermore, the inducible promoters, including ethanol-induced P-ICL1, rhamnose-induced P-LRA3 and P-LRA4, and a bidirectional promoter (P-Mal-P-Per) that is strongly induced by sucrose, further expanded the promoter toolbox in O. polymorpha. Finally, a series of hybrid promoters were constructed via engineering upstream activation sequence (UAS), which increased the activity of native promoter P-LRA3 by 4.7-10.4 times without obvious leakage expression. Therefore, this study provided a group of constitutive, inducible, and hybrid promoters for metabolic engineering of O. polymorpha, and also a feasible strategy for rationally regulating the promoter strength.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据