Immobilization of Recombinant CGTase JCGT8-5 on Magnetically-Modified Silicates and Natural Supports

V. Ivanova; K. Petrov; M. Safarikova; P. Petrova; A. Tonkova; N. Delchev

doi:10.15866/ireche.v6i1.1153

Immobilization of Recombinant CGTase JCGT8-5 on Magnetically-Modified Silicates and Natural Supports

V. Ivanova^(1*), K. Petrov⁽²⁾, M. Safarikova⁽³⁾, P. Petrova⁽⁴⁾, A. Tonkova⁽⁵⁾, N. Delchev⁽⁶⁾

^(*) Corresponding author

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Abstract

Recombinant cyclodextrin glucanotransferase JCGT8-5 (CGTase, EC 2.4.1.19) was effectively immobilized by adsorption on magnetically-modified particles. Silicates (montmorillonite and halloysite), natural supports (oat straw, wheat bran, banana peel, sawdust, hazelnut and peanut shell, coffee beans, tea leaves, algae Chlorella vulgaris) and activated carbon were engaged as carriers, as the most of them were used for the first time for enzyme adsorption. The immobilization capacities of the carriers entrapping ultra-concentrate or purified enzyme were determined. Binding yields reached from 50% to 90% of the initial enzyme quantity. The analysis of the formed cyclodextrins (CDs) revealed that the highest amount of total CDs was obtained by the use of halloysite particles (10.1 mg/ml), followed by sawdust (8.6 mg/ml), algae (8.6 mg/ml) and peanut shell (8.2 mg/ml). A high degree of starch conversion into CDs, ranging from 21 to 25% was achieved for 20 min starch hydrolysis. Variations in γ-CD: α-CD: β-CD ratios due to the immobilization on the different carriers were observed. The six-fold reuse of the magnetic biocatalysts containing purified recombinant CGTase bound to halloysite, washed algae or sawdust provided 29-36 mg/ml CD yield without presence of α-CD for 120 min starch hydrolysis
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Keywords

Immobilization; Recombinant CGTase; Magnetic Carriers; CDs Production

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