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Isolation of Penicillium chitinases: a comparison of enzymes isolated from supernatant and pelleted fractions
Author(s) -
Barros Matthew D,
Bonetti Sandra J.,
Carsella James S.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a57-c
Subject(s) - chitinase , pellet , centrifugation , chromatography , enzyme , extracellular , penicillium , chemistry , size exclusion chromatography , cell wall , chitin , biochemistry , biology , food science , zoology , chitosan
Fungal cell walls are continuously remodeled by hydrolases such as chitinases that modify and recycle cell wall components. Recently, we characterized a chitinase from a less soluble, pelleted fraction of filtrates obtained from 11 day cultures of Penicillium fellutanum grown in Standard Growth medium. Previously, we isolated an extracellular chitinase from a soluble, supernatant fraction from culture filtrates of P. fellulatum grown under identical conditions. These investigations present a comparison of the characteristics of chitinases obtained from the pellet and the supernatant. Chitinase purification is greatly enhanced by centrifugation, which results in an increased concentration of the enzyme activities in the pellet. Both chitinases were purified by multiple steps including ion exchange chromatography and size exclusion chromatography. SDS‐PAGE analyses revealed that the purified pellet chitinase contained major protein bands corresponding to 170 and 150 kD. SDS‐PAGE analysis of the supernatant chitinase fractions revealed major bands corresponding to 81 and 56 kD. Supported by NIH/NIGMS MBRS Grant# 2 S06 GM008197.