Preparation and Properties of Cross-linked Enzyme Aggregates of Trehalose Synthase

doi:10.7506/spkx1002-6630-201309024

Abstract

Abstract:

This paper describes the preparation and properties of cross-linked enzyme aggregates (CLEAs) of trehalose
synthase. The effects of different precipitants, enzyme-to-precipitant concentration ratio, cross-linking intensity and NaBH4
treatment on the activity of CLEAs were investigated as well as their structural features and reactivity. The optimum conditions
for CLEAs preparation were achieved when 10 mg/mL trehalose synthase solution was added with 30 mg/mL of PEG, crosslinked
with 0.5% glutaraldehyde at ambient temperature for 2 h, followed by NaBH4 reduction. CLEAs revealed optimum
reaction temperature of 70 ℃, 20 ℃ higher than that of the free enzyme, and optimum pH of 7.0. Compared to the free enzyme,
CLEAs indicated improved heat and pH stability. Moreover, the tolerance to some metal ions such as Zn2+, Cu2+, Fe2+ and Al3+
was remarkably enhanced. The microscopic morphology of CLEAs showed individual aggregates of 0.2—0.5 μm in particle
size and numerous clusters of different sizes in a “scaffolding” structure.

Key words: trehalose synthase (Tres), cross-linked enzyme aggregates (CLEAs), carrier-free immobilization, cluster

CLC Number:

Q814.2

CHEN Ying1，XIAO Chen-peng2，CHEN Xiao-yun1，YANG Li-wei1，TANG Liu1，LI Ming-chun2，ZHANG Jun1,*. Preparation and Properties of Cross-linked Enzyme Aggregates of Trehalose Synthase[J]. FOOD SCIENCE, 2013, 34(9): 108-113.

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