Expression, characterization, and biochemical properties of recombinant human salivary amylase.
Human salivary amylase, a major component of human salivary secretions, possesses multiple functions in the oral cavity. It is the only enzyme in saliva capable of degrading oligosaccharides, which are used by the oral microflora for nutritional purposes. In order to understand its role in disease processes such as caries, we have undertaken the structure-function analyses of amylase. In this regard, the nonglycosylated human salivary amylase was expressed in a baculovirus expression system. The native and the recombinant amylases exhibit similar biochemical as well as biophysical properties. Unlike recombinant human pancreatic amylase, recombinant human salivary amylase is not glycosylated when expressed in a baculovirus system as determined from the crystal structure determination of the recombinant enzyme. Therefore, this system is suitable for further structure-function work without resorting to enzymatic removal of the carbohydrate chain. Details of the expression, purification, and biophysical properties will be presented.
Ragunath
Chandran
C
Dental Research Center, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103, USA.
Sundar
Krishnan
K
Ramasubbu
Narayanan
N
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000681:Amylases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016367:Baculoviridae; D001483:Base Sequence; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides; D000681:Amylases