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Biosynthesis and characterization of a recombinant eukaryotic allophycocyanin using prokaryotic accessory enzymes.

ABSTRACT: Phycobiliproteins (PBPs) are colored fluorescent proteins present in cyanobacteria, red alga, and cryptophyta. These proteins have many potential uses in biotechnology going from food colorants to medical applications. Allophycocyanin, the simplest PBP, is a heterodimer of ?? subunits that oligomerizes as a trimer (??)3 . Each subunit contains a phycocyanobilin, bound to a cysteine residue, which is responsible for its spectroscopic properties. In this article, we are reporting the expression of recombinant allophycocyanin (rAPC) from the eukaryotic red algae Agarophyton chilensis in Escherichia coli, using prokaryotic accessory enzymes to obtain a fully functional rAPC. Three duet vectors were used to include coding sequences of ? and ? subunits from A. chilensis and accessorial enzymes (heterodimeric lyase cpc S/U, heme oxygenase 1, phycocyanobilin oxidoreductase) from cyanobacteria Arthrospira maxima. rAPC was purified using several chromatographic steps. The characterization of the pure rAPC indicates very similar spectroscopic properties, ?max Abs , ?max Em , fluorescence lifetime, and chromophorylation degree, with native allophycocyanin (nAPC) from A. chilensis. This method, to produce high-quality recombinant allophycocyanin, can be used to express and characterize other macroalga phycobiliproteins, to be used for biotechnological or biomedical purposes.

SUBMITTER: Dagnino-Leone J

PROVIDER: S-EPMC7066465 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Biosynthesis and characterization of a recombinant eukaryotic allophycocyanin using prokaryotic accessory enzymes.

Dagnino-Leone Jorge J Figueroa Maximiliano M Uribe Elena E Hinrichs María Victoria MV Ortiz-López Diego D Martínez-Oyanedel José J Bunster Marta M

MicrobiologyOpen 20200122 3

Phycobiliproteins (PBPs) are colored fluorescent proteins present in cyanobacteria, red alga, and cryptophyta. These proteins have many potential uses in biotechnology going from food colorants to medical applications. Allophycocyanin, the simplest PBP, is a heterodimer of αβ subunits that oligomerizes as a trimer (αβ)<sub>3</sub> . Each subunit contains a phycocyanobilin, bound to a cysteine residue, which is responsible for its spectroscopic properties. In this article, we are reporting the ex ...[more]

PMID: 31970933

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Dataset Information

Biosynthesis and characterization of a recombinant eukaryotic allophycocyanin using prokaryotic accessory enzymes.

Publications

Biosynthesis and characterization of a recombinant eukaryotic allophycocyanin using prokaryotic accessory enzymes.

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