OPTIMIZATION OF CYCLODEXTRIN-?PRODUCTION FROM SAGO STARCH USING RECOMBINANT CYCLODEXTRIN GLYCOCYLTRANSFERASE FROM BACILLUS SP. A2-5A

• Main Author: Novaryatiin, Susi
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/66663
• Institution: Institut Teknologi Bandung

Cyclodextrin (CD) is a cyclic oligosaccharide molecule which is classified into three types: CD ?, ?, and ?, each consists of 6, 7, and 8 glucose units. CDs are produced enzymatically using starch as substrate catalyzed by CD glycosyltransferase (CGTase). This research aimed to determine optimum condition of CD-?production from sago starch using recombinant CGTase (rCGTase) from Bacillus sp. A2-5a, and using the optimum condition, CD-? production was done at 100 mL scale using complexing agent. In this study, rCGTase was purified using Ni-NTA affinity chromatography with gradual elution and concentrated using Nanosep column (10 kDa cutoff). The purified rCGTase was 76.39 kDa in size and displayed ?-cyclization and hydrolysis activities using zimography. The results showed that optimum conditions for CD-?production are concentration of pregelatinized sago starch of 0.5 % (w/v); unit enzyme of 2.6x10 - 2 U, reaction time of 8 hours and concentration of complexing agent N,Ndimethylformamide (DMF) of 1% (v/v). On the addition of DMF 1% (v/v) repeatedly at 100 mL scale production, the highest concentration of CD-?was obtained at reaction time of 8 hours. This research reports for the first time the optimum condition of CD-?production at 100 mL scale using rCGTase from Bacillus sp. A2-5a with DMF as complexing agent.