TY - JOUR
T1 - Expression, characterization, and biochemical properties of recombinant human salivary amylase
AU - Ragunath, Chandran
AU - Sundar, Krishnan
AU - Ramasubbu, Narayanan
N1 - Funding Information:
This study was supported by USPHS Grant DE12585. The authors thank Dr. Hong Li, Department of Biochemistry and Molecular Biology, for the mass spectral data.
PY - 2002/3
Y1 - 2002/3
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0036514613&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036514613&partnerID=8YFLogxK
U2 - 10.1006/prep.2001.1559
DO - 10.1006/prep.2001.1559
M3 - Article
C2 - 11858714
AN - SCOPUS:0036514613
SN - 1046-5928
VL - 24
SP - 202
EP - 211
JO - Protein Expression and Purification
JF - Protein Expression and Purification
IS - 2
ER -