Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 160, Issue -, Pages 20-28Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2019.03.012
Keywords
Astaxanthin; Escherichia colt; Multiple promoters
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Funding
- Ministry of Education, Taiwan, R.O.C. under the ATU plan
- Ministry of Science and Technology (MOST), Taiwan, R.O.C. [106-2311-B-214-001-, 106-2632-B-214-001-]
- Institute of Plant and Microbial Biology, and Scientific Instrument Center, Academia Sinica
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Astaxanthin possesses various biological properties and is used in the animal and fish feed, food, and beverage industries. In this study, we derived zeaxanthin biosynthesis genes (crtE, crtB, crtI, crtY, and crtZ) from Erwinia uredovora and crtW from Agrobacterium aurantiacum. We fused inducible and constitutive promoters to astaxanthin biosynthesis genes to construct a novel plasmid (dubbed PTP3-6) that can effectively enhance free-form astaxanthin (FFAX) production. The PTP3-6 plasmid contains one T7 promoter, driving IPTG inducible crtW expression, and three constitutive promoters (isolated from E. uredovora) driving expression of the other zeaxanthin biosynthesis genes. Escherichia colt BL21 (DE3) cells carrying the PTP3-6 plasmid produced 8.3 mg/g dry cell weight astaxanthin, which is 69.4-fold higher than has been previously reported. Using multiple promoter fusions of astaxanthin biosynthesis genes could be applied in other hosts to enhance astaxanthin production. FFAX was identified in recombinant E. colt cells through ultra-performance liquid chromatography-mass spectrometry.
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