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Purification and characterization of a levanbiose‐producing levanase from Pseudomonas sp. No. 43
Author(s) -
Jung Kang Eun,
Lee Sang Ok,
Lee Jae Dong,
Lee Tae Ho
Publication year - 1999
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1999.tb00558.x
Subject(s) - isoelectric point , chemistry , chromatography , hydrolysis , potassium permanganate , enzyme , pseudomonas , molecular mass , biochemistry , bacteria , biology , organic chemistry , genetics
A levanbiose‐accumulating levanase from Pseudomonas sp. No. 43 was purified to a homogeneous state by (NH 4 ) 2 SO 4 fractionation and by chromatography on DEAE‐Toyopearl 650 M and phenyl‐Toyopearl 650 M columns. The molecular mass and isoelectric point of the enzyme were estimated to be 36 kDa and 5.7 respectively; the optimal pH and temperature for the enzyme reaction were pH 7.0 and 40 °C respectively. The purified enzyme was stable in the pH range 6.0–8.0 at 20 °C and stable up to 50 °C at pH 7.0. The enzyme's activity was inhibited by MnCl 2 , CoCl 2 , AlCl 3 , EDTA and potassium permanganate. The levanase was specific towards the 2,6‐β‐D‐fructosidic linkages of levan and did not hydrolyse other polysaccharides among those examined. The enzyme is an exohydrolase of levan and produced levanbiose as a sole product; the limits of hydrolysis of levans from Zymomonas mobilis and Serratia sp. were 65% and 80% respectively.