Thermostabilization of proteins by diglycerol phosphate, a new compatible solute from the hyperthermophile Archaeoglobus fulgidus.
Diglycerol phosphate accumulates under salt stress in the archaeon Archaeoglobus fulgidus (L. O. Martins, R. Huber, H. Huber, K. O. Stetter, M. S. da Costa, and H. Santos, Appl. Environ. Microbiol. 63:896-902, 1997). This solute was purified after extraction from the cell biomass. In addition, the optically active and the optically inactive (racemic) forms of the compound were synthesized, and the ability of the solute to act as a protecting agent against heating was tested on several proteins derived from mesophilic or hyperthermophilic sources. Diglycerol phosphate exerted a considerable stabilizing effect against heat inactivation of rabbit muscle lactate dehydrogenase, baker's yeast alcohol dehydrogenase, and Thermococcus litoralis glutamate dehydrogenase. Highly homologous and structurally well-characterized rubredoxins from Desulfovibrio gigas, Desulfovibrio desulfuricans (ATCC 27774), and Clostridium pasteurianum were also examined for their thermal stabilities in the presence or absence of diglycerol phosphate, glycerol, and inorganic phosphate. These proteins showed different intrinsic thermostabilities, with half-lives in the range of 30 to 100 min. Diglycerol phosphate exerted a strong protecting effect, with approximately a fourfold increase in the half-lives for the loss of the visible spectra of D. gigas and C. pasteurianum rubredoxins. In contrast, the stability of D. desulfuricans rubredoxin was not affected. These different behaviors are discussed in the light of the known structural features of rubredoxins. The data show that diglycerol phosphate is a potentially useful protein stabilizer in biotechnological applications.
Lamosa
P
P
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2780-156 Oeiras, Portugal.
Burke
A
A
Peist
R
R
Huber
R
R
Liu
M Y
MY
Silva
G
G
Rodrigues-Pousada
C
C
LeGall
J
J
Maycock
C
C
Santos
H
H
D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000187:drug effects; D000818:Animals; D019611:Archaeoglobus fulgidus / Q000737:chemistry* / Q000502:physiology; D018533:Biomass; D003001:Cloning, Molecular; D003013:Clostridium / Q000378:metabolism; D003901:Desulfovibrio / Q000378:metabolism; D004355:Drug Stability; D004795:Enzyme Stability; D004798:Enzymes / Q000737:chemistry* / Q000187:drug effects; D005969:Glutamate Dehydrogenase / Q000737:chemistry / Q000187:drug effects; D005990:Glycerol / Q000494:pharmacology; D005994:Glycerophosphates / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D006358:Hot Temperature; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000187:drug effects; D018482:Muscle, Skeletal / Q000201:enzymology; D010710:Phosphates / Q000494:pharmacology; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000187:drug effects; D012416:Rubredoxins / Q000737:chemistry* / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D019712:Thermococcus / Q000201:enzymology
D004798:Enzymes; D005994:Glycerophosphates; D010710:Phosphates; D011994:Recombinant Proteins; D012416:Rubredoxins; C409185:diglycerol phosphate; D000426:Alcohol Dehydrogenase; D007770:L-Lactate Dehydrogenase; D005969:Glutamate Dehydrogenase; D005990:Glycerol