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A novel and efficient method for the immobilization of thermolysin using sodium chloride salting-in and consecutive microwave irradiation.

Bioresource technology (2011-12-14)
Feifei Chen, Fangkai Zhang, Fangchuan Du, Anming Wang, Weifang Gao, Qiuyan Wang, Xiaopu Yin, Tian Xie
ABSTRACT

Sodium chloride salting-in and microwave irradiation were combined to drive thermolysin molecules into mesoporous support to obtain efficiently immobilized enzyme. When the concentration of sodium chloride was 3 M and microwave power was 40 W, 93.2% of the enzyme was coupled to the support by 3 min, and the maximum specific activity of the immobilized enzyme was 17,925.1 U mg(-1). This was a 4.5-fold increase in activity versus enzyme immobilized using conventional techniques, and a 1.6-fold increase versus free enzyme. Additionally, the thermal stability of the immobilized thermolysin was significantly improved. When incubated at 70°C, there was no reduction in activity by 3.5h, whereas free thermolysin lost most of its activity by 3h. Immobilization also protected the thermolysin against organic solvent denaturation. The microwave-assisted immobilization technique, combined with sodium chloride salting-in, could be applied to other sparsely soluble enzymes immobilization because of its simplicity and high efficiency.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2-Methyl-2-butanol, ReagentPlus®, 99%
Sigma-Aldrich
2-Methyl-2-butanol, ReagentPlus®, ≥99%
Sigma-Aldrich
2-Methyl-2-butanol, anhydrous, ≥99%
Supelco
2-Methyl-2-butanol, analytical standard