Highly Stable, Cold-Active Aldehyde Dehydrogenase from the Marine Antarctic Flavobacterium sp. PL002

• Published in: Fermentation
• Main Authors: Georgiana Necula-Petrareanu, Paris Lavin, Victoria Ioana Paun, Giulia Roxana Gheorghita, Alina Vasilescu, Cristina Purcarea
• Format: Article in Journal/Newspaper
• Language: English
• Published: MDPI AG 2021
• Subjects: aldehyde dehydrogenase; cold-active enzyme; Flavobacterium; Antarctic bacteria; thermostable catalyst; Fermentation industries. Beverages. Alcohol; TP500-660; Antarctic; Antarc*
• Online Access: https://doi.org/10.3390/fermentation8010007; https://doaj.org/article/9f86342f87544e938f5d1f47b8f8f64d

Stable aldehyde dehydrogenases (ALDH) from extremophilic microorganisms constitute efficient catalysts in biotechnologies. In search of active ALDHs at low temperatures and of these enzymes from cold-adapted microorganisms, we cloned and characterized a novel recombinant ALDH from the psychrotrophic Flavobacterium PL002 isolated from Antarctic seawater. The recombinant enzyme (F-ALDH) from this cold-adapted strain was obtained by cloning and expressing of the PL002 aldH gene (1506 bp) in Escherichia coli BL21(DE3). Phylogeny and structural analyses showed a high amino acid sequence identity (89%) with Flavobacterium frigidimaris ALDH and conservation of all active site residues. The purified F-ALDH by affinity chromatography was homotetrameric, preserving 80% activity at 4 °C for 18 days. F-ALDH used both NAD + and NADP + and a broad range of aliphatic and aromatic substrates, showing cofactor-dependent compensatory K M and k cat values and the highest catalytic efficiency (0.50 µM −1 s −1 ) for isovaleraldehyde. The enzyme was active in the 4–60 °C-temperature interval, with an optimal pH of 9.5, and a preference for NAD + -dependent reactions. Arrhenius plots of both NAD(P) + -dependent reactions indicated conformational changes occurring at 30 °C, with four(five)-fold lower activation energy at high temperatures. The high thermal stability and substrate-specific catalytic efficiency of this novel cold-active ALDH favoring aliphatic catalysis provided a promising catalyst for biotechnological and biosensing applications.