Overexpression and Characterization of a Novel Plant Carotenoid Cleavage Dioxygenase 1 from Morus notabilis.

Autor: Qi Z; Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, 159 Long Pan Road, Nanjing, 210037, China.; College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China., Fan X; College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China., Zhu C; College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China., Chang D; College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China., Pei J; College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China.; Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University, 159 Long Pan Road, Nanjing, 210037, China., Zhao L; Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, 159 Long Pan Road, Nanjing, 210037, China.; College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China.
Jazyk: angličtina
Zdroj: Chemistry & biodiversity [Chem Biodivers] 2022 Feb; Vol. 19 (2), pp. e202100735. Date of Electronic Publication: 2021 Dec 29.
DOI: 10.1002/cbdv.202100735
Abstrakt: Synthesis of β-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 β-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 β-ionone, 3-hydroxy-4-oxo-β-ionone, 3-hydroxy-β-ionone, and 3-hydroxy-α-ionone in vitro. MnCCD1 could also cleave lycopene and β-carotene at the 9, 10 (9', 10') bind bond to produce pseudoionone and β-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 % ethanol could increase enzyme activity at above 15 %. However, an obvious decline was observed on enzyme activity as the concentration of Fe 2+ increased (0-1 mM). The V max for β-apo-8'-carotenal was 72.5 U/mg, while the K m 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.
(© 2021 Wiley-VHCA AG, Zurich, Switzerland.)
Databáze: MEDLINE
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