Colorimetric dimethyl sulfide sensor using Rhodovulum sulfidophilum cells based on intrinsic pigment conversion by CrtA.
A colorimetric whole-cell sensor for dimethyl sulfide (DMS) was constructed based on the in vivo conversion of intrinsic pigments in response to the analyte. In a marine bacterium, Rhodovulum sulfidophilum, carotenoids are synthesized via the spheroidene pathway. In this pathway, demethylspheroidene, a yellow carotenoid, is converted to spheroidene under catalysis of O-methyltransferase. Spheroidene monooxygenase (CrtA) catalyzes the terminal step of the pathway and converts spheroidene to spheroidenone, a red carotenoid. Here, the CrtA gene in R. sulfidophilum was removed and then reintroduced downstream of the DMS dehydrogenase gene promoter. Using this whole-cell sensor, 3 muM DMS or dimethyl sulfoxide can be detected without adding any color-forming reagent. The ratio of the red spheroidenone to total carotenoids increased, as the DMS concentration was raised to 0.3 mM. Comparison of the signal to the background color indicated a shift in the color coordinate from a yellow to a red hue. An intense signal was obtained with 1-day incubation at a high cell density when sensor cells at the exponential growth phase were used. These results show that the genetically engineered R. sulfidophilum cells can be used to monitor the quality of marine aquacultural environments by the naked eye.
Maeda
Isamu
I
Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Japan.
Yamashiro
Hidenori
H
Yoshioka
Daiki
D
Onodera
Masanori
M
Ueda
Shunsaku
S
Kawase
Masaya
M
Miyasaka
Hitoshi
H
Yagi
Kiyohito
K
D015374:Biosensing Techniques / Q000379:methods*; D002338:Carotenoids / Q000378:metabolism*; D003124:Colorimetry / Q000379:methods*; D004121:Dimethyl Sulfoxide / Q000032:analysis; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D041903:Rhodovulum / Q000235:genetics*; D013440:Sulfides / Q000032:analysis*
D012333:RNA, Messenger; D013440:Sulfides; C005637:spheroidene; C104880:spheroidenone; D002338:Carotenoids; C004784:dimethyl sulfide; D004121:Dimethyl Sulfoxide