Comparative proteomic and metabolomic analysis of Streptomyces tsukubaensis reveals the metabolic mechanism of FK506 overproduction by feeding soybean oil.
FK506 (tacrolimus) is a 23-membered polyketide macrolide that possesses powerful immunosuppressant activity. In this study, feeding soybean oil into the fermentation culture of Streptomyces tsukubaensis improved FK506 production by 88.8%. To decipher the overproduction mechanism, comparative proteomic and metabolomic analysis was carried out. A total of 72 protein spots with differential expression in the two-dimensional gel electrophoresis (2-DE) were identified by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF-MS), and 66 intracellular metabolites were measured by gas chromatography-mass spectrometer (GC-MS). The analysis of proteome and metabolome indicated that feeding soybean oil as a supplementary carbon source could not only strengthen the FK506 precursor metabolism and energy metabolism but also tune the pathways related to transcriptional regulation, translation, and stress response, suggesting a better intracellular metabolic environment for the synthesis of FK506. Based on these analyses, 20 key metabolites and precursors of FK506 were supplemented into the soybean oil medium. Among them, lysine, citric acid, shikimic acid, and malonic acid performed excellently for promoting the FK506 production and biomass. Especially, the addition of malonic acid achieved the highest FK506 production, which was 1.56-fold of that in soybean oil medium and 3.05-fold of that in initial medium. This report represented the first comprehensive study on the comparative proteomics and metabolomics applied in S. tsukubaensis, and it would be a rational guidance to further strengthen the FK506 production.
Key Laboratory of System Bioengineering (Tianjin University), Ministry of Education, 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, 300072, People's Republic of China.
Jun
J
Wang
Key Laboratory of System Bioengineering (Tianjin University), Ministry of Education, 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, 300072, People's Republic of China.
Huanhuan
H
Liu
TEDA School of Biological Sciences and Biotechnology, Nankai University, TEDA, Tianjin, 300457, People's Republic of China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Nankai University, Tianjin, 300074, People's Republic of China.
Di
D
Huang
Key Laboratory of System Bioengineering (Tianjin University), Ministry of Education, 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, 300072, People's Republic of China.
Lina
L
Jin
Key Laboratory of System Bioengineering (Tianjin University), Ministry of Education, 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, 300072, People's Republic of China.
Cheng
C
Wang
Key Laboratory of System Bioengineering (Tianjin University), Ministry of Education, Tianjin, 300072, People's Republic of China. jpwen@tju.edu.cn.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China. jpwen@tju.edu.cn.
Jianping
J
Wen
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D019149:Bioreactors; D019343:Citric Acid / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005285:Fermentation; D008401:Gas Chromatography-Mass Spectrometry; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D063990:Gene Ontology; D007166:Immunosuppressive Agents / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism; D008314:Malonates / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000235:genetics; D055432:Metabolomics; D058977:Molecular Sequence Annotation; D040901:Proteomics; D012765:Shikimic Acid / Q000378:metabolism; D013024:Soybean Oil / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D016559:Tacrolimus / Q000378:metabolism*
D001426:Bacterial Proteins; D007166:Immunosuppressive Agents; D008314:Malonates; D019343:Citric Acid; D012765:Shikimic Acid; D013024:Soybean Oil; C030290:malonic acid; D008239:Lysine; D016559:Tacrolimus
Comparative proteomics; FK506; Metabolomics; Soybean oil; Streptomyces tsukubaensis