Pressure-enhanced activity and stability of a hyperthermophilic protease from a deep-sea methanogen.

Applied and Environmental Microbiology
P C Michels, Douglas S Clark

Abstract

We describe the properties of a hyperthermophilic, barophilic protease from Methanococcus jannaschii, an extremely thermophilic deep-sea methanogen. This enzyme is the first protease to be isolated from an organism adapted to a high-pressure-high-temperature environment. The partially purified enzyme has a molecular mass of 29 kDa and a narrow substrate specificity with strong preference for leucine at the P1 site of polypeptide substrates. Enzyme activity increased up to 116(deg)C and was measured up to 130(deg)C, one of the highest temperatures reported for the function of any enzyme. In addition, enzyme activity and thermostability increased with pressure: raising the pressure to 500 atm increased the reaction rate at 125(deg)C 3.4-fold and the thermostability 2.7-fold. Spin labeling of the active-site serine revealed that the active-site geometry of the M. jannaschii protease is not grossly different from that of several mesophilic proteases; however, the active-site structure may be relatively rigid at moderate temperatures. The barophilic and thermophilic behavior of the enzyme is consistent with the barophilic growth of M. jannaschii observed previously (J. F. Miller et al., Appl. Environ. Microbiol. 54:3039-3042, 1988).

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Citations

Jan 4, 2012·Extremophiles : Life Under Extreme Conditions·Adrienne KishAndrew Steele
May 25, 1999·Protein Science : a Publication of the Protein Society·M M SunD S Clark
Mar 10, 2001·Microbiology and Molecular Biology Reviews : MMBR·C Vieille, G J Zeikus
Jan 31, 2013·Chemistry : a European Journal·Anoklase J-L AyitouJ Sivaguru
Jun 26, 2013·Extremophiles : Life Under Extreme Conditions·Eiji OhmaeChiaki Kato
Oct 21, 2014·Environmental Microbiology·María AlcaideUNKNOWN MAMBA Consortium
Sep 7, 2000·Microbiology and Molecular Biology Reviews : MMBR·A T BullM Goodfellow
Jan 19, 2002·The Journal of Biological Chemistry·David G KellnerStephen G Sligar
Mar 12, 2017·Extremophiles : Life Under Extreme Conditions·Debamitra ChakravortySanjukta Patra
Aug 14, 2019·International Journal of Biological Macromolecules·Yanan Li, Haibin Zhang
May 2, 2002·Biochimica Et Biophysica Acta·Boonchai B BoonyaratanakornkitDouglas S Clark
Dec 10, 1999·Microbiology and Molecular Biology Reviews : MMBR·A J MacarioE Conway de Macario
Aug 5, 2010·Journal of Agricultural and Food Chemistry·Bong-Kuk KoCherl-Ho Lee
Oct 20, 2005·Journal of Bioscience and Bioengineering·Takeo ImaiMasood Ahmed Siddiqui
Jun 20, 2006·Journal of Biotechnology·Francesca SimonatoDouglas H Bartlett
May 23, 2015·The Journal of Physical Chemistry. B·Paolo A CalligariGerald R Kneller

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