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Psychrophilic enzymes: hot topics in cold adaptation

Nature Reviews Microbiology volume 1, pages 200–208 (2003)Cite this article

Key Points

  • More than three-quarters of the Earth's surface is cold — oceans with a constant temperature of 4–5°C below a depth of 1,000m cover approximately 70% of the Earth's surface. The microorganisms that occupy these regions are known as psychrophiles. To maintain essential chemical reactions at these temperatures, psychrophilic enzymes are cold active and heat labile.

  • Psychrophilic enzymes maintain high activity at low temperatures mainly by decreasing the temperature dependence of the reaction that is catalysed. This is achieved by improving the mobility or flexibility of the active site. As a consequence, substrate binding is generally less efficient, but specific mutations can compensate for this adaptive drift, especially when substrate binding (Km) has a regulatory function.

  • The catalytic centre of cold-active enzymes is identical to that of mesophilic enzymes, to maintain specificity, but local interactions might help to improve catalysis at low temperatures, such as better accessibility to the active site or favourable electrostatic interactions with the substrate. Generally, adaptive mutations favouring active-site flexibility are located outside the catalytic centre. All known interaction types that stabilize a protein are reduced in number and strength, but each enzyme family uses one or a combination of the altered interactions to gain in molecular mobility.

  • At least in the case of the best-studied psychrophilic enzyme (chitobiase), the relationships between stability and activity at low temperatures have been shown by site-directed mutagenesis. Stabilizing the psychrophilic enzyme, by engineering the weak interactions found in the mesophilic enzyme, decreases activity and improves substrate binding of the mutants.

  • The stability curves of psychrophilic enzymes reveal several unsuspected properties. They are optimally stable at room temperature, which reflects the dominant effect of hydrophobic forces in protein folding. However, they are cold labile and more prone to cold denaturation than mesophilic proteins, which is a phenomenon that might set a biophysical lower limit to life at low temperatures. In addition, the thermodynamic contributions to their stability are the opposite to that of mesophilic proteins, for example the stability of cold-active enzymes is entropy-driven at low temperatures.

  • Directed evolution experiments show that several molecular adjustments can lead to cold activity. However, in cold environments, the simplest strategy seems to be to lose stability, in the absence of selection for stable proteins, to gain in flexibility and activity, under a strong selective pressure for cold-active enzymes.

Abstract

More than three-quarters of the Earth's surface is occupied by cold ecosystems, including the ocean depths, and polar and alpine regions. These permanently cold environments have been successfully colonized by a class of extremophilic microorganisms that are known as psychrophiles (which literally means cold-loving). The ability to thrive at temperatures that are close to, or below, the freezing point of water requires a vast array of adaptations to maintain the metabolic rates and sustained growth compatible with life in these severe environmental conditions.

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Figure 1: Temperature dependence of enzyme activity.
Figure 2: Inactivation of psychrophilic enzymes.
Figure 3: Kinetic optimization of Km.
Figure 4: Thermal unfolding of DNA ligases.
Figure 5: Stability curves of psychrophilic (blue), mesophilic (yellow) and thermophilic (red) proteins.
Figure 6: Mutational analysis.

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Acknowledgements

Research in the authors' laboratory is supported by the European Union, the Région Wallonne (Belgium), the Fonds National de la Recherche Scientifique (Belgium) and the University of Liége. The facilities offered by the Institut Polaire Français are also acknowledged.

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  1. Laboratory of Biochemistry, Institute of Chemistry B6, University of Liége, Liége-Sart Tilman, B-4000, Belgium

    Georges Feller & Charles Gerday

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  1. Georges Feller
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  2. Charles Gerday
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Correspondence to Georges Feller.

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FURTHER INFORMATION

Astrobiology

Basic concepts of extremophiles

Georges Feller and Charles Gerday's laboratory

Marine microbiology

Glossary

k cat

The catalytic constant is the maximal enzyme reaction rate at a given temperature, which is expressed as the number of substrate molecules that are transformed by one molecule of enzyme per unit of time. It is also known as the turnover number.

K m

This is the substrate concentration that is required to produce 50% of the maximal activity. In a simple reaction mechanism, this parameter reflects the enzyme affinity for the substrate (if the association/dissociation of the enzyme–substrate complex is fast in respect to product formation).

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Feller, G., Gerday, C. Psychrophilic enzymes: hot topics in cold adaptation. Nat Rev Microbiol 1, 200–208 (2003). https://doi.org/10.1038/nrmicro773

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