Engineering of Synechococcus sp. strain PCC 7002 for the photoautotrophic production of light-sensitive riboflavin (vitamin B2) |
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| Institution: | 1. Laboratory for Bioinformatics, Graduate School of Systems Life Sciences, Kyushu University, 804 Westwing, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan;2. NODAI Genome Research Center, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan;3. Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8603, Japan;4. Kobe University Research Center for Inland Seas, 2746 Iwaya, Awaji, Hyogo 656-2401, Japan;5. Department of Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan;1. DBT-ICT Centre for Energy Biosciences, Institute of Chemical Technology, Matunga, Mumbai 400019, India;2. Department of Chemical Engineering, Institute of Chemical Technology, Matunga, Mumbai 400019, India;3. Chemical Engineering Department, School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, AZ 85287, USA;1. College of Environmental Science & Engineering, Nankai University, Tianjin 300071, China;2. Department of Earth & Atmospheric Sciences, University of Alberta, Edmonton, Alberta T6G 2E3, Canada;3. Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada;4. UMR6538 Domaines Océaniques, CNRS-Université de Bretagne Occidentale, Institut Universitaire Européen de la Mer, 29280 Plouzané, France;5. Institut für Geo- und Umweltnaturwissenschaften, Mineralogie-Petrologie Albert-Ludwigs-Universität, 79104 Freiburg, Germany |
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| Abstract: | Due to their capability of photosynthesis and autotrophic growth, cyanobacteria are currently investigated with regard to the sustainable production of a wide variety of chemicals. So far, however, no attempt has been undertaken to engineer cyanobacteria for the biotechnological production of vitamins, which is probably due to the light-sensitivity of many of these compounds. We now describe a photoautotrophic bioprocess to synthesize riboflavin, a vitamin used as a supplement in the feed and food industry. By overexpressing the riboflavin biosynthesis genes ribDGEABHT from Bacillus subtilis in the marine cyanobacterium Synechococcus sp. PCC 7002 riboflavin levels in the supernatant of the corresponding recombinant strain increased 56-fold compared to the wild-type. Introduction of a second promoter region upstream of the heterologous ribAB gene – coding for rate-limiting enzymatic functions in the riboflavin biosynthesis pathway – led to a further increase of riboflavin levels (211-fold compared to the wild-type). Degradation of the light-sensitive product riboflavin was prevented by culturing the genetically engineered Synechococcus sp. PCC 7002 strains in the presence of dichromatic light generated by red light-emitting diodes (λ = 630 and 700 nm). Synechococcus sp. PCC 7002 naturally is resistant to the toxic riboflavin analog roseoflavin. Expression of the flavin transporter pnuX from Corynebacterium glutamicum in Synechococcus sp. PCC 7002 resulted in roseoflavin-sensitive recombinant strains which in turn could be employed to select roseoflavin-resistant, riboflavin-overproducing strains as a chassis for further improvement. |
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| Keywords: | Photoautotrophy Riboflavin Roseoflavin |
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