Lysine acetylation can generate highly charged enzymes with increased resistance toward irreversible inactivation.
This paper reports that the acetylation of lysine epsilon-NH3(+) groups of alpha-amylase--one of the most important hydrolytic enzymes used in industry--produces highly negatively charged variants that are enzymatically active, thermostable, and more resistant than the wild-type enzyme to irreversible inactivation on exposure to denaturing conditions (e.g., 1 h at 90 degrees C in solutions containing 100-mM sodium dodecyl sulfate). Acetylation also protected the enzyme against irreversible inactivation by the neutral surfactant TRITON X-100 (polyethylene glycol p-(1,1,3,3-tetramethylbutyl)phenyl ether), but not by the cationic surfactant, dodecyltrimethylammonium bromide (DTAB). The increased resistance of acetylated alpha-amylase toward inactivation is attributed to the increased net negative charge of alpha-amylase that resulted from the acetylation of lysine ammonium groups (lysine epsilon-NH3(+) --> epsilon-NHCOCH3). Increases in the net negative charge of proteins can decrease the rate of unfolding by anionic surfactants, and can also decrease the rate of protein aggregation. The acetylation of lysine represents a simple, inexpensive method for stabilizing bacterial alpha-amylase against irreversible inactivation in the presence of the anionic and neutral surfactants that are commonly used in industrial applications.
Shaw
Bryan F
BF
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Schneider
Gregory F
GF
Bilgiçer
Basar
B
Kaufman
George K
GK
Neveu
John M
JM
Lane
William S
WS
Whitelegge
Julian P
JP
Whitesides
George M
GM
D000107:Acetylation; D001407:Bacillus / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D019075:Electrophoresis, Capillary; D004795:Enzyme Stability / Q000187:drug effects; D008239:Lysine / Q000378:metabolism*; D017830:Octoxynol / Q000494:pharmacology; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization; D000516:alpha-Amylases / Q000737:chemistry / Q000378:metabolism*
D001426:Bacterial Proteins; D000644:Quaternary Ammonium Compounds; D012967:Sodium Dodecyl Sulfate; D017830:Octoxynol; D000516:alpha-Amylases; C022806:dodecyltrimethylammonium; D008239:Lysine