CAS OpenIR
Metabolic engineering of Cupriavidus necator H16 for improved chemoautotrophic growth and PHB production under oxygen-limiting conditions
Tang, Ruohao1,2; Weng, Caihong1,2; Peng, Xiaowei1; Han, Yejun1
2020-09-01
Source PublicationMETABOLIC ENGINEERING
ISSN1096-7176
Volume61Pages:11-23
AbstractThe oxygen-limiting condition promotes the accumulation of ployhydroxybutyrate (PHB) in C. necator H16, while the growth of which is restricted. Under autotrophic culture using carbon dioxide, hydrogen, and oxygen as substrates, the oxygen concentration below 6.9% (v/v) in the mixture is considered as a safe condition. It also expected to achieve cell rapid growth and large accumulation of PHB simultaneously under the oxygen-limiting condition in C. necator H16. In this study, a metabolically engineered strain capable of both rapid growth and large accumulation of PHB under oxygen-limiting conditions was constructed based on the transcriptomic analysis. In the comparative transcriptomic analysis, the genes related to energy-generating of C. necator H16 at autotrophic culture were downregulated under oxygen-limiting conditions (3%, v/v). Besides, the genes related to the key intermediates (pyruvate and acetyl-CoA) metabolism in PHB biosynthetic pathway were analyzed. Most of which were downregulated, except the genes ldh, iclA, and ackA2 respectively encoding L-lactate dehydrogenase, isocitrate lyase, and acetate kinase were upregulated under oxygen-limiting conditions (3%, v/v). The Vitreoscilla hemoglobin (VHb) has the ability to promote aerobic metabolism and energy generation. To promote the bacterium growth and improve the energy generation in C. necator H16 under oxygen-limiting conditions, the VHb gene was introduced into C. necator H16 with the optimized promoter PphaC1-j5. Moreover, VHb was localized to the periplasmic space of the bacterium by the traction of membrane-bound hydrogenase (MBH) signal peptide. By optimizing the knockout of different genes, it was found that knockout of ldh can improve PHB production and reduce the by-products. Finally, a recombinant strain Reh01 (p2M-pj-v) was constructed by heterologous expression of vgb and ldh knockout in C. necator H16. Compared with the control (Reh (p2)) under oxygen-limiting conditions (3%, v/v), the dry cell weight (DCW), PHB content, and PHB production of Reh01 (p2M-pj-v) increased by 31.0%, 30.9%, and 71.5%, respectively. From the perspectives of transcriptome and metabolic engineering, the work provides new ideas to achieve rapid cell growth and large PHB accumulation in C. necator under oxygen-limiting and autotrophic conditions.
KeywordCupriavidus necator H16 Carbon dioxide Oxygen-limiting conditions Transcription Metabolic engineering Ployhydroxybutyrate
DOI10.1016/j.ymben.2020.04.009
Language英语
WOS KeywordVITREOSCILLA HEMOGLOBIN ; GENE-EXPRESSION ; HYDROGEN BACTERIA ; TANDEM PROMOTERS ; CELL-DENSITY ; CO2 ; CULTIVATION ; EXCRETION ; CULTURE ; O-2
Funding ProjectNational Natural Science Foundation of China[21676279] ; National Key Research and Development Project of China[2019YFB1503800]
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
Funding OrganizationNational Natural Science Foundation of China ; National Key Research and Development Project of China
WOS IDWOS:000570475100002
PublisherACADEMIC PRESS INC ELSEVIER SCIENCE
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/42162
Collection中国科学院过程工程研究所
Corresponding AuthorPeng, Xiaowei; Han, Yejun
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Sch Chem Engn, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Tang, Ruohao,Weng, Caihong,Peng, Xiaowei,et al. Metabolic engineering of Cupriavidus necator H16 for improved chemoautotrophic growth and PHB production under oxygen-limiting conditions[J]. METABOLIC ENGINEERING,2020,61:11-23.
APA Tang, Ruohao,Weng, Caihong,Peng, Xiaowei,&Han, Yejun.(2020).Metabolic engineering of Cupriavidus necator H16 for improved chemoautotrophic growth and PHB production under oxygen-limiting conditions.METABOLIC ENGINEERING,61,11-23.
MLA Tang, Ruohao,et al."Metabolic engineering of Cupriavidus necator H16 for improved chemoautotrophic growth and PHB production under oxygen-limiting conditions".METABOLIC ENGINEERING 61(2020):11-23.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Tang, Ruohao]'s Articles
[Weng, Caihong]'s Articles
[Peng, Xiaowei]'s Articles
Baidu academic
Similar articles in Baidu academic
[Tang, Ruohao]'s Articles
[Weng, Caihong]'s Articles
[Peng, Xiaowei]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Tang, Ruohao]'s Articles
[Weng, Caihong]'s Articles
[Peng, Xiaowei]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.