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Retinal biosynthesis in Eubacteria: in vitro characterization of a novel carotenoid oxygenase from Synechocystis sp. PCC 6803

Sandra Ruch

Albert‐Ludwigs University of Freiburg, Institut for Biology II, Cellbiology, Schaenzlestr. 1, D‐79104 Freiburg, Germany.

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Peter Beyer

Albert‐Ludwigs University of Freiburg, Institut for Biology II, Cellbiology, Schaenzlestr. 1, D‐79104 Freiburg, Germany.

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Hansgeorg Ernst

BASF Aktiengesellschaft, Fine Chemicals, and Biocatalysis Research, GVF‐B009, D‐67056 Ludwigshafen, Germany.

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Salim Al‐Babili

Corresponding Author

Albert‐Ludwigs University of Freiburg, Institut for Biology II, Cellbiology, Schaenzlestr. 1, D‐79104 Freiburg, Germany.

E‐mail

salim.albabili@biologie.uni‐freiburg.de

; Tel. (+49) 761 203 2529, Fax: (+49) 761 203 2675.
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First published: 12 January 2005
Cited by: 62

Summary

Retinal and its derivatives represent essential compounds in many biological systems. In animals, they are synthesized through a symmetrical cleavage of β‐carotene catalysed by a monooxygenase. Here, we demonstrate that the open reading frame sll1541 from the cyanobacterium Synechocystis sp. PCC 6803 encodes the first eubacterial, retinal synthesizing enzyme (Diox1) thus far reported. In contrast to enzymes from animals, Diox1 converts β‐apo‐carotenals instead of β‐carotene into retinal in vitro. The identity of the enzymatic product was proven by HPLC, GC‐MS and in a biological test. Investigations, of the stereospecifity showed that Diox1 cleaved only the all‐trans form of β‐apo‐8′‐carotenal, yielding all‐trans‐retinal. However, Diox1 exhibited wide substrate specificity with respect to chain‐lengths and functional end‐groups. Although with divergent Km and Vmax values, the enzyme converted β‐apo‐carotenals, (3R)‐3‐OH‐β‐apo‐carotenals as well as apo‐lycopenals into retinal, (3R)‐3‐hydroxy‐retinal and acycloretinal respectively. In addition, the alcohols of these substrates were cleaved to yield the corresponding retinal derivatives.

Number of times cited: 62

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