Single-step purification of chitosanases from Bacillus cereus using expanded bed chromatography

Single-step purification of chitosanases from Bacillus cereus using expanded bed chromatography

A chitosanase-producing strain was isolated and identified as Bacillus cereus C-01. The purification and characterization of two chitosanases were studied. The purification assay was accomplished by ion exchange expanded-bed chromatography. Experiments were carried out in the presence and in the abs...

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Bibliographic Details
Published in:International journal of biological macromolecules Vol. 82; pp. 291 - 298
Main Authors: Araújo, Nathália Kelly de, Pagnoncelli, Maria Giovana Binder, Pimentel, Vanessa Carvalho, Xavier, Maria Luiza Oliveira, Padilha, Carlos Eduardo Araújo, Macedo, Gorete Ribeiro de, Santos, Everaldo Silvino dos
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 01-01-2016
Subjects:
B. cereus
Bacillus cereus - enzymology
Bioseparation
Chitosan
Chitosanase
Chromatography - methods
Enzyme Activation
Expanded bed adsorption
Glycoside Hydrolases - chemistry
Glycoside Hydrolases - isolation & purification
Hydrogen-Ion Concentration
Ion exchange
Ions - chemistry
Temperature
B. cereus
Chitosan
Ion exchange
Expanded bed adsorption
Chitosanase
Bioseparation
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Summary:A chitosanase-producing strain was isolated and identified as Bacillus cereus C-01. The purification and characterization of two chitosanases were studied. The purification assay was accomplished by ion exchange expanded-bed chromatography. Experiments were carried out in the presence and in the absence of cells through different expansion degree to evaluate the process performance. The adsorption experiments demonstrated that the biomass does not affect substantially the adsorption capacity of the matrix. The enzyme bound to the resin with the same extent using clarified and unclarified broth (0.32 and 0.30U/g adsorbent, respectively). The fraction recovered exhibited 31% of the yield with a 1.26-fold increase on the specific activity concerned to the initial broth. Two chitosanases from different elution steps were recovery. Chit A and Chit B were stable at 30–60°C, pH 5.5–8.0 and 5.5–7.5, respectively. The highest activity was found at 55°C, pH 5.5 to Chit A and 50°C, pH 6.5 to Chit B. The ions Cu2+, Fe2+ and Zn2+ indicated inhibitory effect on chitosanases activities that were significantly activated by Mn2+. The methodology applied in this study enables the partial purification of a stable chitosanase using a feedstock without any pre-treatment using a single-step purification.
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content type line 23
ISSN:0141-8130
1879-0003
DOI:10.1016/j.ijbiomac.2015.09.063