Downstream processing of inulinase. Comparison of different techniques

Area of Science:

• Biotechnology
• Enzyme Technology
• Microbial Fermentation

Background:

• Inulinase is a valuable enzyme used in various industrial processes.
• Efficient recovery and purification of inulinase are crucial for cost-effective applications.
• Candida kefyr is a known source of inulinase, but its recovery needs optimization.

Purpose of the Study:

• To investigate and compare different methods for the recovery and purification of inulinase from Candida kefyr.
• To determine the concentration factor (C(f)) and enrichment factor (E(f)) for each method.
• To identify the most effective method for industrial-scale inulinase purification.

Main Methods:

• Cultivation of Candida kefyr DSM 70106 in a medium with inulin as the carbon source.
• Enzyme recovery using cross-flow filtration (microfiltration, cell diafiltration, enzyme ultrafiltration, and diafiltration).
• Enzyme recovery using liquid-liquid extraction with N-Benzyl-N-Dodecyl-N-bis[2-hydroxyethyl] ammonium chloride (BDBAC) reversed micelles.
• Enzyme recovery using expanded-bed adsorption.

Main Results:

• Approximately 92% of the inulinase was recovered directly from the medium.
• Cross-flow filtration achieved a concentration factor (C(f)) of 7.5 and an enrichment factor (E(f)) of 2.2.
• Liquid-liquid extraction yielded C(f) = 2.5 and E(f) = 2.7.
• Expanded-bed adsorption resulted in C(f) = 2.8 and the highest E(f) = 4.3.

Conclusions:

• Expanded-bed adsorption is the most effective method for inulinase purification, achieving the highest enrichment factor.
• The study demonstrates the feasibility of efficient inulinase recovery from Candida kefyr.
• Optimized enzyme recovery methods can enhance the industrial viability of inulinase applications.