Synthesis of cinnabarinic acid by metabolically engineered Pseudomonas chlororaphis GP72. Issue 11 (26th July 2019)

• Record Type: Journal Article
• Authors: Yue, Sheng‐Jie; Song, Chen; Li, Song; Huang, Peng; Guo, Shu‐Qi; Hu, Hong‐Bo; Wang, Wei; Zhang, Xue‐Hong
• Is Part Of: Biotechnology and bioengineering. Volume 116:Issue 11(2019)
• Page Start: 3072
• Page End: 3083
• Publication Date: 2019-07-26
• Subjects: cinnabarinic acid -- DHHA -- Pseudomonas chlororaphis GP72 -- renewable bioresources -- synthetic pathway; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.27118
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: Cinnabarinic acid is a valuable phenoxazinone that has broad applications in the pharmaceutical, chemical, and dyeing industries. However, few studies have investigated the production of cinnabarinic acid or its derivatives using genetically engineered microorganisms. Herein, an efficient synthetic pathway of cinnabarinic acid was designed and constructed in Pseudomonas chlororaphis GP72 for the first tim, which was more straightforward and robust than the known eukaryotic biosynthetic pathways. First, we screened and identified trans ‐2, 3‐dihydro‐3‐hydroxyanthranilic acid (DHHA) dehydrogenases from Escherichia coli MG1655 (encoded by entA ), Streptomyces sp. NRRL12068 (encoded by bomO ) and Streptomyces chartreusis NRRL3882 (encoded by calB 3 ) based on the structural similarity of the substrate and product, and the DHHA dehydrogenase encoded by calB 3 was selected for the synthesis of cinnabarinic acid due to its high DHHA conversion rate. Subsequently, cinnabarinic acid was synthesized by the expression of the DHHA dehydrogenase CalB3 and the phenoxazinone synthase CotA in the DHHA‐producing strain P. chlororaphis GP72, resulting in a cinnabarinic acid titer of 20.3 mg/L at 48 hr. Further fermentation optimization by the addition of Cu 2+, H2 O2, and with adding glycerol increased cinnabarinic acid titer to 136.2 mg/L in shake flasks. The results indicate that P. chlororaphis GP72 may be engineered as a microbial cell factory to produce cinnabarinic acid or its derivatives from renewable bioresources. Abstract : Cinnabarinic acid is a valuable phenoxazinone that has broad applications in the pharmaceutical, chemical, and dyeing industries. However, few studies have investigated the production of cinnabarinic acid or its derivatives using genetically engineered microorganisms.