Phycobilin heterologous production from the Rhodophyta Porphyridium cruentum.

Autor: Montoya EJO; Laboratorio de Biotecnología, Sede de Investigación Universitaria - SIU, Universidad de Antioquia, Carrera 53 # 61 - 30 - SIU. Torre 1 Laboratorio de 210, Medellín 050010, Colombia. Electronic address: erika.obando@udea.edu.co., Dorion S; Institut de Recherche en Biologie Végétale, Université de Montréal, 4101 Rue Sherbrooke est, Montréal, QC H1X 2B2, Canada., Atehortua-Garcés L; Laboratorio de Biotecnología, Sede de Investigación Universitaria - SIU, Universidad de Antioquia, Carrera 53 # 61 - 30 - SIU. Torre 1 Laboratorio de 210, Medellín 050010, Colombia., Rivoal J; Institut de Recherche en Biologie Végétale, Université de Montréal, 4101 Rue Sherbrooke est, Montréal, QC H1X 2B2, Canada.
Jazyk: angličtina
Zdroj: Journal of biotechnology [J Biotechnol] 2021 Nov 20; Vol. 341, pp. 30-42. Date of Electronic Publication: 2021 Sep 06.
DOI: 10.1016/j.jbiotec.2021.09.001
Abstrakt: Phycobiliproteins are colored, active molecules with potential use in different industries. They are the union of proteins and bilins (Chromophores). The primary source of phycobiliproteins is algae; however, the traditional algae culture has production restrictions. The production in bacterial models can be a more efficient alternative to produce these molecules. However, the lack of knowledge in some steps of the phycobiliprotein metabolic pathway limits this alternative. Porphyridium cruentum is a single cell red alga with a high phycobiliprotein content. Its protein sequences were the basis for phycobilin production in this study. In this study, we cloned and characterized enzymes presumably involved in the chromophore production of P. cruentum. Using sequences obtained from its transcriptome, we characterized two cDNA sequences predicted to code respectively for a ferredoxin-dependent bilin reductase and a bilin lyase-isomerase. We expressed these enzymes in Escherichia coli to obtain in vivo evidence of their enzymatic activity on the substrate biliverdin IXα. Lastly, we analyzed them using thin-layer chromatography, spectrophotometry, and fluorescence spectroscopy. These experiments provided evidence of bilin modification. The expressed bilin lyase-isomerase did not show significant activity over the biliverdin molecule. On the contrary, the expressed ferredoxin-dependent bilin reductase showed activity over the biliverdin.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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