Metabolic Engineering of the Chl d-Dominated Cyanobacterium Acaryochloris marina: Production of a Novel Chl Species by the Introduction of the Chlorophyllide a Oxygenase Gene

• Published in: Plant and cell physiology Vol. 53; no. 3; pp. 518 - 527
• Main Authors: Tsuchiya, Tohru, Mizoguchi, Tadashi, Akimoto, Seiji, Tomo, Tatsuya, Tamiaki, Hitoshi, Mimuro, Mamoru
• Format: Journal Article
• Language: English
• Published: Japan Oxford University Press 01.03.2012
• Subjects: Acaryochloris marina; Biosynthetic Pathways - genetics; Chlorophyll - chemistry; Chlorophyll - metabolism; Chromatography, High Pressure Liquid; Conjugation, Genetic; Cyanobacteria; Cyanobacteria - cytology; Cyanobacteria - enzymology; Cyanobacteria - genetics; Expression vectors; Gene transfer; Genes, Bacterial - genetics; Genetic Vectors - genetics; Host Specificity - genetics; Light effects; Mass spectroscopy; metabolic engineering; Metabolic Engineering - methods; N.M.R; Optical properties; Oxygenase; Oxygenases - genetics; Oxygenases - metabolism; Pigments; Plasmids; Plasmids - genetics; Prochlorothrix - enzymology; Reproducibility of Results; Spectroscopy; Spectrum Analysis; Transformation; Transformation, Genetic; Metabolic engineering; Chlorophyllide; oxygenase; Transformation; Novel Chl
• ISSN: 0032-0781; 1471-9053
• DOI: 10.1093/pcp/pcs007

In oxygenic photosynthetic organisms, the properties of photosynthetic reaction systems primarily depend on the Chl species used. Acquisition of new Chl species with unique optical properties may have enabled photosynthetic organisms to adapt to various light environments. The artificial production of a new Chl species in an existing photosynthetic organism by metabolic engineering provides a model system to investigate how an organism responds to a newly acquired pigment. In the current study, we established a transformation system for a Chl d-dominated cyanobacterium, Acaryochloris marina, for the first time. The expression vector (constructed from a broad-host-range plasmid) was introduced into A. marina by conjugal gene transfer. The introduction of a gene for chlorophyllide a oxygenase, which is responsible for Chl b biosynthesis, into A. marina resulted in a transformant that synthesized a novel Chl species instead of Chl b. The content of the novel Chl in the transformant was approximately 10% of the total Chl, but the level of Chl a, another Chl in A. marina, did not change. The chemical structure of the novel Chl was determined to be [7-formyl]-Chl d P by mass spectrometry and nuclear magnetic resonance spectroscopy. [7-Formyl]-Chl d P is hypothesized to be produced by the combined action of chlorophyllide a oxygenase and enzyme(s) involved in Chl d biosynthesis. These results demonstrate the flexibility of the Chl biosynthetic pathway for the production of novel Chl species, indicating that a new organism with a novel Chl might be discovered in the future.