Purification of beta-amylase from alfalfa (Medicago sativa L.) seeds.
An amylase from alfalfa (Medicago sativa L. c.v. Moapa) seeds was purified by column chromatography and gel filtration, followed by chromatofocusing on Mono P HR 5/20. The last step was effective for separation of the alfalfa amylase to a homogeneous state. The purified amylase was identified as beta-amylase from the fact that only beta-maltose was formed by the enzymatic degradation of soluble starch. The molecular weight and specific activity of the beta-amylase (E1%(280 nm) = 18.3) were determined to be 61,000 and 1,077 A.U./mg, respectively. The beta-amylase activity was inhibited by the modification of sulfhydryl groups with p-chloromercuribenzoic acid. The optimum pH and isoelectric point of alfalfa beta-amylase were 7.0 and 4.8, respectively, which were different from other plant beta-amylases.
Kohno
A
A
Department of Utilization of Biological Resources, Graduate School of Science and Technology, Kobe University, Hyogo.
Nanmori
T
T
Shinke
R
R
D000681:Amylases / Q000302:isolation & purification*; D002241:Carbohydrates; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D000455:Medicago sativa / Q000201:enzymology*; D008970:Molecular Weight; D009904:Optical Rotation; D012639:Seeds / Q000201:enzymology*; D001614:beta-Amylase / Q000302:isolation & purification* / Q000378:metabolism
D002241:Carbohydrates; D000681:Amylases; D001614:beta-Amylase