PARTIAL PURIFICATION AND CHARACTERIZATION OF -GLUCOSIDASE FROM ALOE VERA L. LEAVES

Year 2009, Issue: 40, 65 - 75, 27.04.2011

Tuğba Yılmaz Ayşe Can

Abstract

The partial purification of β-glucosidase from the pulp of Aloe vera L. Burm. fil.
(sarisabir) leaves and some of its kinetic properties is presented. The fresh leaves of
A. vera were used; the gel portion was separated and the remaining leaf pulps were cut
into small pieces. The crude extract was prepared by homogenization of the leaf pulps in
phosphate buffered saline (PBS), pH 7.4 and subsequent centrifugation. β-Glucosidase
active fraction was precipitated by 30%-65% ammonium sulphate from the crude
extract. Hydroxylapatite column chromatography resulted in a single peak showing
β-glucosidase activity eluted with 200 mM phosphate buffer. The partially purified
enzyme showed two protein and a single activity band in polyacrylamide gel
electrophoresis. It was found that the enzyme exhibited maximum activity at 50oC and
at pH 4.4. Km and Vmax values for 4-nitrophenyl-β-D-glucopyranoside were 6.8x10-4
M and 4,58x10-3 U, respectively. When β-glucosidase activity was investigated
throughout the year, it was found that the activity increased in winter and decreased in
summer.

Keywords

glycosidases, -glucosidase, Aloe vera, purification, seasonal variation

PARTIAL PURIFICATION AND CHARACTERIZATION OF -GLUCOSIDASE FROM ALOE VERA L. LEAVES

Abstract

Aloe vera L. Burm. fil. (sarısabır) yapraklarının pulpa kısmından β-glukozidaz
kısmen saflaştırıldı ve bazı kinetik özellikleri incelendi. A.vera‟nın taze yaprakları kullanıldı; jel kısmı ayrıldı ve kalan yaprak pulpaları küçük parçalara kesildi. Yaprak
pulpalarının fosfatla tamponlanmış %0.9 NaCl (PBS), pH 7.4 ile homojenizasyonunun
ardından santrifüje edilmesiyle ham ekstre hazırlandı. β-Glukozidaz aktivitesi gösteren
fraksiyon ham ekstreden %30-%65 amonyum sülfat ile çöktürüldü. Hidroksilapatit
kolon kromatografisi sonucunda 200 mM fosfat tamponu ile β-glukozidaz aktivitesi
gösteren tek bir pik elde edildi. Kısmen saflaştırılan enzim poliakrilamid jel
elektroforezinde iki protein ve bir aktivite bandı gösterdi. Enzimin en yüksek aktiviteyi
50C‟de ve pH 4.4‟te gösterdiği ve 4-nitrofenil-β-D-glukopiranozide karşı Km
değerinin 6.8x10-4 M, Vmax değerinin ise 4.58x10-3 U olduğu saptandı. β-Glukozidaz
aktivitesi yıl boyunca incelendiğinde aktivitenin kış aylarında arttığı, yaz aylarında ise
azaldığı bulundu.

Keywords

glycosidases, -glucosidase, Aloe vera, purification, seasonal variation

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