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RINA IMANIAR
School of Pharmacy, Institut Teknologi bandung, Jalan Ganesha 10, Bandung 40132, West Java, Indonesia
Biochemistry, Genetics & Molecular Biology Immunology & microbiology Medicine & Pharmacology

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Enzymatic Characterization of Recombinant Cyclodextrin Glycosyltransferase from Bacillus sp. a2-5a using Sagoo Starch as Substrate RINA IMANIAR; CATUR RIANI; DESSY NATALIA; DEBBIE SOFFIE RETNONINGRUM
Microbiology Indonesia Vol. 6 No. 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (485.698 KB) | DOI: 10.5454/mi.6.3.5

Abstract

Cyclodextrin (CD) is a cyclic oligosaccharide molecule and depending on the number of glucose molecules, three types of CDs are commonly used, α-CD, β-CD, and γ-CD. CDs can be produced enzymatically using starch as substrate catalyzed by CD glycosyltransferase (CGTase). In current research, recombinant CGTase production from the synthetic gene was optimized for its production using three growth media and two induction temperatures. The highest yield was obtained in Luria Bertani medium at 25 °C. The rCGTase protein was affinity purified as a 76.39 kDa protein which showed α-cyclization and starch hydrolysis activities using zymography method. The optimum temperature, pH and incubation time was 55 °C, 6, and 24 h, respectively. The enzyme was stable at a wide pHs in the range of 5-10, retained its half activity at 56 °C for 30 min and had cyclization ratio for α-CD : β-CD : γ-CD was 4 : 81 : 15. An amount of 542 mg β-cyclodextrin was produced from 100 mL reaction of 1% (b/v) sagoo starch using 38.4 μg rCGTase in optimum condition. This work reports for the first time the character of rCGTase from Bacillus sp. A2-5a using sagoo starch as a substrate.
Co-Authors DEBBIE SOFFIE RETNONINGRUM DESSY NATALIA Riani, Catur