Bacterial Two Component Systems: Overexpression and Purification: In Vitro and In Vivo Inhibitor Screens

Dietrich, Alina; Gajdiss, Mike; Türck, Michael; Monk, Ian R.; Bierbaum, Gabriele

doi:10.1007/978-1-0716-2855-3_17

Cited by 2 publications

(2 citation statements)

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“…Recently, an optimized protocol for the quantification of proteins with a maximum of two transmembrane helices was published. 45 An alternative protein quantification method is the split GFP assays 46 that were so far not applied to transmembrane proteins.…”

Section: ■ Discussionmentioning

confidence: 99%

“…Given the structural differences between the studied lycopene β-cyclases, a relative comparison or a reliable quantification remains a challenge. Recently, an optimized protocol for the quantification of proteins with a maximum of two transmembrane helices was published . An alternative protein quantification method is the split GFP assays that were so far not applied to transmembrane proteins.…”

Section: Discussionmentioning

confidence: 99%

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Screening of Structurally Distinct Lycopene β-Cyclases for Production of the Cyclic C40 Carotenoids β-Carotene and Astaxanthin by Corynebacterium glutamicum

Göttl

Pucker

Wendisch

et al. 2023

J. Agric. Food Chem.

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Lycopene β-cyclase (EC 5.5.1.19) is one of the key enzymes in the biosynthesis of β-carotene and derived carotenoids. It catalyzes isomerase reactions to form β-carotene from lycopene by β-cyclization of both of its ψ-ends. Lycopene β-cyclases are widespread in nature. We systematically analyzed the phylogeny of lycopene β-cyclases from all kingdoms of life and predicted their transmembrane structures. To this end, a collection of previously characterized lycopene β-cyclase polypeptide sequences served as bait sequences to identify their closest homologues in a range of bacteria, archaea, fungi, algae, and plant species. Furthermore, a DeepTMHMM scan was applied to search for the presence of transmembrane domains. A phylogenetic tree suggests at least five distinct clades, and the DeepTMHMM scan revealed that lycopene β-cyclases are a group of structurally different proteins: membrane-bound and cytosolic enzymes. Representative lycopene β-cyclases were screened in the lycopene-overproducing Corynebacterium glutamicum strain for β-carotene and astaxanthin production. This systematic screening facilitates the identification of new enzymes for carotenoid production. Higher astaxanthin production and less reduction of total carotenoids were achieved with the cytosolic lycopene β-cyclase CrtL from Synechococcus elongatus and the membrane-bound heterodimeric lycopene β-cyclase CrtYcd from Brevibacterium linens.

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Section: ■ Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%