TY - JOUR PY - 2014/10/01 Y2 - 2024/03/29 TI - Optimization of β-Cyclodextrin Production from Sago Starch using Recombinant Cyclodextrin Glycocyltransferase from Bacillus sp. A2-5a JF - Microbiology Indonesia JA - Microbiol indones VL - 8 IS - 3 SE - Articles DO - 10.5454/mi.8.3.6 UR - https://jurnal.permi.or.id/index.php/mionline/article/view/290 SP - 6 AB - <p>Cyclodextrin (CD) is a cyclic oligosaccharide molecule which is classified into three types: α-CD, β-CD, and γ-CD, each consists of 6, 7, and 8 glucose units. CDs are produced enzymatically using starch as the substrate catalyzed by CD glycosyltransferase (CGTase). This research was aimed to determine optimum condition of β-CD production from sago starch using recombinant CGTase (rCGTase) from <em>Bacillus </em>sp. A2-5a, and using the optimum condition, β-CD production was done at 100 mL scale using complexing agents. In this study, rCGTase was overproduced in <em>Escherichia coli</em> BL21(DE3) and purified applying Ni-NTA affinity chromatography with gradual elution of imidazole and concentrated using Nanosep column. The purified rCGTase was 76.39 kDa in size and displayed β-cyclization and hydrolysis activities using zymography. The results showed that optimum conditions for β-CD production was achieved when preheated sago starch of 0.5% (w/v), enzyme of 2.6x10<sup>-2</sup> U, and N,N-dimethylformamide (DMF) as complexing agent of 1% (v/v) were used and the reaction time was 8 h. When DMF of 1% (v/v) was added repeatedly at 100 mL scale production, the highest concentration of β-CD was obtained at the reaction time of 8 h. This research reported for the first time the optimum condition of b-CD production at 100 mL scale using rCGTase from <em>Bacillus</em> sp. A2-5a with DMF as complexing agent.</p> ER -