Biochemical characterization of alpha-amylase from the yeast Cryptococcus flavus.

Abstract:

:During our screening of amylolytic microorganisms from Brazilian fruits, we isolated a yeast strain classified as Cryptococcus flavus. When grown on starch-containing medium this strain exhibited the highest amylase production after 24 h of cultivation. The extracellular amylase from C. flavus was purified from the culture broth by a single step using chromatography on a Sephacryl S-100 column. The enzyme was purified 16.14-fold with a yield of 50.21% of the total activity. The purified enzyme was a glycoprotein with an apparent molecular mass of 75 and 84.5 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively. The enzyme lost approximately 50% of the molecular mass after treatment with glycosidases. The major end products of starch, amylose, amylopectin, pullulan and glycogen were maltose and maltotriose. The K(m) value for the pure enzyme was 0.056 mg ml(-1) with soluble starch as the substrate. Enzyme activity was optimal at pH 5.5 and 50 degrees C. The enzyme retained 90% of the activity after incubation at 50 degrees C for 60 min and was inhibited by Cu(2+), Fe(2+) and Hg(2+).

journal_name

FEMS Microbiol Lett

authors

Wanderley KJ,Torres FA,Moraes LM,Ulhoa CJ

doi

10.1016/S0378-1097(03)00955-8

keywords:

subject

Has Abstract

pub_date

2004-02-16 00:00:00

pages

165-9

issue

2

eissn

0378-1097

issn

1574-6968

pii

S0378109703009558

journal_volume

231

pub_type

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