Purification, biochemical characterization, and implications of an alkali-tolerant catalase from the spacecraft-associated and oxidation-resistant Acinetobacter gyllenbergii 2P01AA

• Published in: Astrobiology Vol. 15; no. 4; p. 291
• Main Authors: Muster, N, Derecho, I, Dallal, F, Alvarez, R, McCoy, K B, Mogul, R
• Format: Journal Article
• Language: English
• Published: United States 01.04.2015
• Subjects: Acinetobacter - enzymology; Alkalies - pharmacology; Amino Acid Sequence; Catalase - antagonists & inhibitors; Catalase - chemistry; Catalase - isolation & purification; Catalase - metabolism; Electrophoresis, Polyacrylamide Gel; Enzyme Stability - drug effects; Hydrogen Peroxide - pharmacology; Hydrogen-Ion Concentration; Kinetics; Mass Spectrometry; Models, Molecular; Molecular Sequence Data; Oxidation-Reduction - drug effects; Sequence Homology, Amino Acid; Sodium Hydroxide - pharmacology; Spacecraft; Structural Homology, Protein; Temperature
• ISSN: 1557-8070
• DOI: 10.1089/ast.2014.1242

Herein, we report on the purification, characterization, and sequencing of catalase from Acinetobacter gyllenbergii 2P01AA, an extremely oxidation-resistant bacterium that was isolated from the Mars Phoenix spacecraft assembly facility. The Acinetobacter are dominant members of the microbial communities that inhabit spacecraft assembly facilities and consequently may serve as forward contaminants that could impact the integrity of future life-detection missions. Catalase was purified by using a 3-step chromatographic procedure, where mass spectrometry provided respective subunit and intact masses of 57.8 and 234.6 kDa, which were consistent with a small-subunit tetrameric catalase. Kinetics revealed an extreme pH stability with no loss in activity between pH 5 and 11.5 and provided respective kcat/Km and kcat values of ∼10(7) s(-1) M(-1) and 10(6) s(-1), which are among the highest reported for bacterial catalases. The amino acid sequence was deduced by in-depth peptide mapping, and structural homology suggested that the catalases from differing strains of A. gyllenbergii differ only at residues near the subunit interfaces, which may impact catalytic stability. Together, the kinetic, alkali-tolerant, and halotolerant properties of the catalase from A. gyllenbergii 2P01AA are significant, as they are consistent with molecular adaptations toward the alkaline, low-humidity, and potentially oxidizing conditions of spacecraft assembly facilities. Therefore, these results support the hypothesis that the selective pressures of the assembly facilities impact the microbial communities at the molecular level, which may have broad implications for future life-detection missions.