title: Integration of parallel13C-labeling experiments and in silico pathway analysis for enhanced production of ascomycin.
abstract: Herein, the hyper-producing strain for ascomycin was engineered based on13C-labeling experiments and elementary flux modes analysis (EFMA). First, the metabolism of non-model organism Streptomyces hygroscopicus var. ascomyceticus SA68 was investigated and an updated network model was reconstructed using13C- metabolic flux analysis. Based on the precise model, EFMA was further employed to predict genetic targets for higher ascomycin production. Chorismatase (FkbO) and pyruvate carboxylase (Pyc) were predicted as the promising overexpression and deletion targets, respectively. The corresponding mutant TD-FkbO and TD-ΔPyc exhibited the consistency effects between model prediction and experimental results. Finally, the combined genetic manipulations were performed, achieving a high-yield ascomycin engineering strain TD-ΔPyc-FkbO with production up to 610 mg/L, 84.8% improvement compared with the parent strain SA68. These results manifested that the integration of13C-labeling experiments and in silico pathway analysis could serve as a promising concept to enhance ascomycin production, as well as other valuable products. Biotechnol. Bioeng. 2017;114: 1036-1044. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, People's Republic of China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, People's Republic of China.
authors.forename: Haishan
authors.initials: H
authors.lastname: Qi
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, People's Republic of China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, People's Republic of China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Lv
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, People's Republic of China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, People's Republic of China.
authors.forename: Kejing
authors.initials: K
authors.lastname: Song
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, People's Republic of China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, People's Republic of China.
authors.forename: Jianping
authors.initials: J
authors.lastname: Wen
mesh_terms: D018533:Biomass; D002247:Carbon Isotopes / Q000032:analysis / Q000378:metabolism*; D003198:Computer Simulation; D005285:Fermentation; D005947:Glucose / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D064688:Metabolic Flux Analysis; D008954:Models, Biological; D015203:Reproducibility of Results; D013302:Streptomyces / Q000378:metabolism*; D016559:Tacrolimus / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; C058028:immunomycin; D005947:Glucose; D016559:Tacrolimus
keywords: 13C-labeling experiments; Streptomyces hygroscopicus var. ascomyceticus; ascomycin; elementary flux modes analysis; metabolic network model