Carotenoid synthesis is probably present in the common ancestor of Bacteria and Archaea; the phytoene synthase gene crtB is universal among carotenoid synthesizers. Among eukaryotes, plants and algae inherited the cyanobacterial pathway via biosynthesis of their plastids, while fungi retain a archaeal-like pathway.[4] Among all these synthesizers, several possible selection and arrangements of biosynthetic genes exist, consisting of one gene cluster cluster, several clusters, or no clustering at all.[5][a]
Role of CRT genes in carotenoid biosynthesis
The CRT gene cluster consists of twenty-five genes such as crtA, crtB, crtC, crtD, crtE, crtF, crtG, crtH, crtI, crtO, crtP, crtR, crtT, crtU, crtV, and crtY, crtZ. These genes play a role in varying stages of the Astaxanthin biosynthesis and Carotenoid biosynthesis (Table 1).[6]
crtE encodes for an enzyme known as geranylgeranyl diphosphate synthase known to catalyze the condensation reaction of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) into geranylgeranyl diphosphate (GGDP).[7][8] Two GGDP molecules are subsequently converted into a single phytoene molecule by phytoene synthase, an enzyme encoded by crtB, known as PSY in Chlorophyta.[2][7][8] The following desaturation of phytoene into ζ-carotene is catalyzed by the phytoene desaturase encoded by crtI, crtP, and/or PDS.[2][7][8] ζ -carotene can also be obtained through phytoene using the carotene 2,4-desaturase enzyme (crtD).[2][9] Depending on the species, varying carotenoids are accumulated following these steps.[1][10]
Spirilloxanthin
Spirilloxanthin is obtained from lycopene following a hydration, desaturation, and methylation reaction. These reactions are catalyzed by carotene hydratase (crtC), carotene 3,4- desaturase (crtD), and carotene methyltransferase (crtF), respectively.[8][1]
Canthaxanthin
Lycopene is cyclized through two enzymes lycopene cyclase and β-C-4-oxygenase/β-carotene ketolase encoded on the crtY (in Chlorophyta) /crtL (in cyanobacteria), and crtW, respectively. crtY cyclizes lycopene into β-carotene, which is subsequently oxygenated by crtW to form canthaxanthin.[8]
Zeaxanthin and lutein
Zeaxanthin and lutein are obtained through hydroxylation of α- and β-carotene.[1] Hydroxylation of Zeaxanthin occurs by β-carotene hydroxylase an enzyme encoded on the crtR (in cyanobacteria) and crtZ gene (in Chlorophyta).[8]
Other
Zeaxanthin can be further processed to obtain zeaxanthin-diglucoside by Zeaxanthin glucosyl transferase (crtX).
Echinenone is obtained from β -carotene through the catalyzing enzyme β-C-4-oxygenase/β-carotene ketolase (crtO).[11]CrtO, also known as bkt2 in Chlorophyta, is also involved in the conversion of other carotenoids into Canthaxanthin, 3-Hydroxyechinenone, 3'-Hydroxyechinenone, Adonixanthin, and Astaxanthin.[11][12]CrtZ, similarly to crtO, is also capable of converting carotenoids into β-cryptoxanthin, Zeaxanthin, 3-Hydroxyechinenone, 3'-Hydroxyechinenone, Astaxanthin, Adonixanthin, and Adonirubin.[11]
crtH catalyzes the isomerization of cis-carotenes into trans-carotenes through carotenoid isomerase.[2]
crtG encodes for carotenoid 2,2'- β-hydroxylase, this enzyme leads to the formation of 2-hydroxylated and 2,2′-dihydroxylated products in E coli.[13]
Table 1: role of CRT genes in carotenoid biosynthesis [2]
Lycopene cyclase (two in Cyanobacteria: crtL-b became plant lcy-B, crtL-e became plant lcy-E)[5]
Cyclization of lycopene
crtD
Carotene 3,4-desaturase
Conversion of phytoene to ζ-carotene
crtA
Spheroidene monooxygenase
Conversion of spheroidene to spheroidenone
crtR+
β-carotene hydroxylase (various Cyanobacteria)
Hydroxylation of β-carotene to zeaxanthin
crtZ*
β-carotene hydroxylase (various Chlorophyta)
Hydroxylation of β-carotene to zeaxanthin
crtX
Zeaxanthin glucosyl transferase
Conversion of zeaxanthin to zeaxanthin-diglucoside
crtW (bkt2*)
β-C-4-oxygenase/β-carotene ketolase
Conversion of β-carotene to canthaxanthin
crtO
β-C-4-oxygenase/β-carotene ketolase
Conversion of β-carotene to echinenone
crtC
Carotene hydratase
Conversion of neurosporene to demethylspheroidene and lycopene to hydroxy derivatives
crtG
Carotenoid 2,2′-β-hydroxylase
Conversion of myxol to 2-hydroxymyxol and zeaxanthin to nostoxanthin
crtK
Carotenoid regulation
-
* In Chlorophyta, + In cyanobacteria
Phylogeny
Previous studies have indicated through phylogenetic analysis that evolutionary patterns of crt genes are characterized by horizontal gene transfer and gene duplication events.[14]
Horizontal gene transfer has been hypothesized to have occurred between cyanobacteria and Chlorophyta, as similarities in these genes have been found across taxa.[14] Note, however, that some cyanobacteria retained their nature. Horizontal gene transfer among species occurred with a high probability in genes involved in the initial steps of the carotenoid biosynthesis pathway such as crtE, crtB, crtY, crtL, PSY, and crtQ. These genes are often well conserved while others involved in the later stages of Carotenoid biosynthesis such as crtW and crtO are less conserved.[1] The less conserved nature of these genes allowed for the expansion of the carotenoid biosynthesis pathway and its end products. Amino acid variations within crt genes have evolved due to purifying and adaptive selection.[2]
Gene duplications are suspected to have occurred due to the presence of multiple copies of ctr clusters or genes within a single species.[2] An example of this can be seen in the Bradyrhizobium ORS278 strain, where initial crt genes can be found (excluding crtC, crtD, and crtF genes) as well as a second crt gene cluster. This second gene cluster has been shown to also be involved in carotenoid biosynthesis using its crtparalogs.[8][15]
References
^For concrete examples of the diversity of gene organization, compare the clusters presented in [6] figure 1 (6 genomes), PMID37887056 figure 1 (4 genomes), PMID22963379 figure 1 (10 genomes), and PMID32155882 figure 3 (11 genomes).
^ abcdeCarotenoid biosynthetic pathway: molecular phylogenies and evolutionary behavior of crt genes in eubacteria. Phadwal K, Gene, 17 January 2005, volume 345, issue 1, pages 35-43, PMID15716108
^ abcdefghMolecular phylogenies and evolution of crt genes in algae. Chen Q, Jiang JG and Wang F, Crit Rev Biotechnol., Apr-Jun 2007;, volume 27, issue 2, pages 77-91, PMID17578704
^Activation and analysis of cryptic crt genes for carotenoid biosynthesis from Streptomyces griseus. Schumann G1, Nürnberger H, Sandmann G and Krügel H, Mol Gen Genet., 28 October 1996, volume 252, issue 6, pages 658-666, PMID8917308
^ abSandmann, G (2021). "Diversity and Evolution of Carotenoid Biosynthesis from Prokaryotes to Plants.". Carotenoids: Biosynthetic and Biofunctional Approaches. Advances in experimental medicine and biology. Vol. 1261. pp. 79–94. doi:10.1007/978-981-15-7360-6_7. PMID33783732.
