Isolation and Functional Characterization of Carotenoid Cleavage Dioxygenase-1 from Laurus nobilis L. (Bay Laurel) Fruits

Abstract

Bay laurel (Laurus nobilis L.) is an agriculturally important tree used in food, drugs, and the cosmetics industry. Many of the health beneficial properties of bay laurel are due to volatile terpene metabolites that they contain, including various norisoprenoids. Despite their importance, little is known about the norisoprenoid biosynthesis in Laurus nobilis fruits. We found that the volatile norisoprenoids 6-methyl-5-hepten-2-one, pseudoionone, and β-ionone accumulated in Laurus nobilis fruits in a pattern reflecting their carotenoid content. A full-length cDNA encoding a potential carotenoid cleavage dioxygenase (LnCCD1) was isolated. The LnCCD1 gene was overexpressed in Escherichia coli, and recombinant protein was assayed for its cleavage activity with an array of carotenoid substrates. The LnCCD1 protein was able to cleave a variety of carotenoids at the 9,10 (9',10') and 5,6 (5',6') positions to produce 6-methyl-5-hepten-2-one, pseudoionone, β-ionone, and α-ionone. Our results suggest a role for LnCCD1 in Laurus nobilis fruit flavor biosynthesis.

Synthesis and Function of Apocarotenoid Signals in Plants

“…In other instances, perhaps CCD1 cleaves carotenoid substrates leaked into the cytosol under particular conditions, such as plastid degradation during fruit ripening. The observed increases in CCD1 expression in the fruits of many species, and the concurrent increases in fruit volatile emission, support this hypothesis [27,86,[90][91][92]. The broad substrate specificity of fruit-specific CCD1 enzymes such as SlCCD1b also supports the notion of CCD1 'indiscriminately' cleaving leaked carotenoids [23].…”

GmCCD4 controls carotenoid content in soybeans

“…In Arabidopsis , four CCDs (CCD1, CCD4, CCD7, CCD8) and five NCEDs (NCED2, NCED3, NCED5, NCED6, NCED9) were identified (Bouvier et al ., 2005). CCD1 and CCD4 cleavage carotenoids to form volatile small apocarotenoids (Hou et al ., 2016; Nacke et al ., 2012; Pu et al ., 2020; Yahyaa et al ., 2015; Zhang et al ., 2015; Zheng et al ., 2019). CCD7/MAX3 and CCD8/MAX4 are involved in the synthesis of strigolactones (SLs) (Alder et al ., 2012; Gomez‐Roldan et al ., 2008; Wang et al ., 2019; Wang et al ., 2020).…”

Efficient production of Pseudoionone with multipathway engineering in Escherichia coli

“…; Yahyaa et al . , ). Currently, there is still no report on de novo production of pseudoionone using a microbial cell factory.…”

“…Significance and Impact of the Study: The present study promises a green and sustainable route for pseudoionone production using a microbial cell factory. (C9-C10 0 ) and/or C5-C6 (C5 0 -C6 0 ) (Ibdah et al 2006;Vogel et al 2008;Yahyaa et al 2013Yahyaa et al , 2015. Currently, there is still no report on de novo production of pseudoionone using a microbial cell factory.…”