Academic Journal
Engineering of Riboflavin Transporter and Enzymes in Bacillus subtilis Facilitates High-Efficiency FAD Synthesis from RF.
| Τίτλος: | Engineering of Riboflavin Transporter and Enzymes in Bacillus subtilis Facilitates High-Efficiency FAD Synthesis from RF. |
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| Συγγραφείς: | Wu Y; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi214122, Jiangsu, China., Sun Y; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi214122, Jiangsu, China., Miao H; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi214122, Jiangsu, China., Xia X; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi214122, Jiangsu, China., Gao L; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi214122, Jiangsu, China. |
| Πηγή: | ACS synthetic biology [ACS Synth Biol] 2026 Aug 21; Vol. 15 (8), pp. 3272-3284. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 101575075 Publication Model: Print Cited Medium: Internet ISSN: 2161-5063 (Electronic) Linking ISSN: 21615063 NLM ISO Abbreviation: ACS Synth Biol Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Washington, D.C. : American Chemical Society, c2012- |
| Ιατρικοί όροι (MeSH): | Bacillus subtilis*/metabolism , Bacillus subtilis*/genetics , Riboflavin*/metabolism , Flavin-Adenine Dinucleotide*/biosynthesis , Flavin-Adenine Dinucleotide*/metabolism , Membrane Transport Proteins*/metabolism , Membrane Transport Proteins*/genetics , Metabolic Engineering*/methods, Bacterial Proteins/metabolism ; Bacterial Proteins/genetics ; Streptomyces/genetics ; Streptomyces/metabolism ; Nucleotidyltransferases/metabolism ; Nucleotidyltransferases/genetics |
| Περίληψη: | The biosynthesis of flavin adenine dinucleotide (FAD), an essential redox coenzyme, is limited by inefficient substrate utilization and low catalytic activity. To address this, an "Uptake-Catalysis" system was constructed in Bacillus subtilis for FAD synthesis from riboflavin (RF). First, the Streptomyces davawensis transporter (RibM) was coexpressed with native FAD synthase (BsFADs) at a 2:1 ratio, which increased RF consumption to 51%. Subsequently, the isothermal compressibility perturbation engineering (ICPE) strategy was adopted, generating the optimum mutant BsFADsP61W+R202Y, which increased catalytic efficiency (kcat/Km) toward RF and FMN by 83% and 73%, respectively. Mechanistic analysis revealed that this improvement was due to a 39% shortening of RF substrate channel. Finally, FAD titer peaked at 563 mg/L with a conversion rate (cr-FAD) of 92% in shaking-flask fermentation optimization. This work achieved a 225-fold increase in FAD production, validating the effectiveness of the uptake-catalysis synergistic strategy, thereby establishing an efficient and scalable synthesis paradigm. (© 2026 American Chemical Society.) |
| Grant Information: | BK20233003 Basic Research Program of Jiangsu Province; 2023YFF1103703 National Key Research and Development Program of China |
| Contributed Indexing: | Keywords: Bacillus subtilis; flavin adenine dinucleotide; isothermal compressibility perturbation engineering strategy; transporter |
| Substance Nomenclature: | TLM2976OFR (Riboflavin) 146-14-5 (Flavin-Adenine Dinucleotide) 0 (Bacterial Proteins) 0 (Membrane Transport Proteins) EC 2.7.7.- (Nucleotidyltransferases) |
| Entry Date(s): | Date Created: 20260511 Date Completed: 20260822 Latest Revision: 20260822 |
| Update Code: | 20260823 |
| DOI: | 10.1021/acssynbio.6c00153 |
| PMID: | 42113034 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 2161-5063 |
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| DOI: | 10.1021/acssynbio.6c00153 |