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245 1 0 _aOverexpression and characterization of a novel plant carotenoid cleavage dioxygenase 1 from Morus notabilis
490 0 _vChemistry & BioDiversity, 19(2), p.e202100735, 2022
520 3 _aSynthesis of ?-ionone in microbial cell factories is limited by the efficiency of carotenoid cleavage dioxygenases (CCDs). To obtain genes responsible for specific cleavage of carotenoids generating ?-ionone, a novel carotenoid cleavage dioxygenase 1 from Morus notabilis was cloned and overexpressed in Escherichia coli. The MnCCD1 protein was able to cleave a variety of carotenoids at the positions 9, 10 (9?, 10?)to produce ?-ionone, 3-hydroxy-4-oxo-?-ionone, 3-hydroxy-?-ionone, and 3-hydroxy-?-ionone in vitro. MnCCD1 could also cleave lycopene and ?-carotene at the 9, 10 (9?, 10?)bind bond to produce pseudoionone and ?-ionone, respectively, in E. coli accumulating carotenoids. The enzyme activity of MnCCD1 was reached 2.98 U/mL at optimized conditions (temperature 28?°C, IPTG 0.1 mM, induction time 24 h). The biochemical characterization of MnCCD1 revealed the optimal activities were at pH 8.4 and 35?°C. The addition of 10? percent ethanol could increase enzyme activity at above 15? percent. However, an obvious decline was observed on enzyme activity as the concentration of Fe2+ increased (0-1 mM). The Vmax for ?-apo-8?-carotenal was 72.5 U/mg, while the Km was 0.83 mM. The results provide a foundation for developing the application of carotenoid cleavage dioxygenases as biocatalysis and synthetic biology platforms to produce volatile aroma components from carotenoids.
650 1 4 _aMORUS NOTABILIS
650 1 4 _aCAROTENOID CLEAVAGE DIOXYGENASES
650 1 4 _aCCD1
650 1 4 _aB-IONONE
700 1 2 _aQi, Z.
700 1 2 _aFan, X.
700 1 2 _aZhu, C.
700 1 2 _aChang, D.
700 1 2 _aPei, J.
700 1 2 _aZhao, L.
856 4 0 _uhttps://drive.google.com/file/d/1ULG5GOTFyrX9NZZ03VyBo5JZk1GahDv8/view?usp=drivesdk
_zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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