Recovery and purification of chitosanase produced by Bacillus cereus using expanded bed adsorption and central composite design

• Published in: Journal of separation science Vol. 39; no. 4; pp. 709 - 716
• Main Authors: de Araújo, Nathália Kelly, Pimentel, Vanessa Carvalho, Silva, Nayane Macedo Portela, Araújo Padilha, Carlos Eduardo, de Macedo, Gorete Ribeiro, dos Santos, Everaldo Silvino
• Format: Journal Article
• Language: English
• Published: Germany Wiley-VCH 01.02.2016; Blackwell Publishing Ltd; Wiley Subscription Services, Inc
• Subjects: Adsorption; Bacillus cereus; Bacillus cereus - enzymology; Bacteriology; Bioseparation; Buffers; Chitosan - chemistry; Chitosanase; chromatography; Chromatography - methods; Distributors; Downstream Processing; Economics; Enzymes; Ethanolamines; Expanded bed adsorption; Glycoside Hydrolases - chemistry; Glycoside Hydrolases - isolation & purification; Hydrodynamics; Hydrogen-Ion Concentration; Industrial Microbiology - methods; Ligands; Molecular Weight; Optimization; Purification; Recovery; Regression Analysis; statistical analysis; Downstream Processing; Expanded bed adsorption; Chitosanase; Bioseparation
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.201500900

This study presents a system for expanded bed adsorption for the purification of chitosanase from broth extract in a single step. A chitosanase‐producing strain was isolated and identified as Bacillus cereus C‐01 and used to produce chitosanases. The expanded bed adsorption conditions for chitosanase purification were optimized statistically using STREAMLINEᵀᴹ DEAE and a homemade column (2.6 × 30.0 cm). Dependent variables were defined by the quality criteria purification factor (P) and enzyme yield to optimize the chromatographic process. Statistical analyses showed that the optimum conditions for the maximum P were 150 cm/h load flow velocity, 6.0 cm settled bed height, and 7.36 cm distributor height. Distributor height had a strong influence on the process, considerably affecting both the P and enzyme yield. Optimizing the purification variables resulted in an approximately 3.66‐fold increase in the P compared with the value under nonoptimized conditions. This system is promising for the recovery of chitosanase from B. cereus C‐01 and is economically viable because it promotes the reduction steps.