Applied Microbiology and Biotechnology

, Volume 51, Issue 4, pp 504–509

Purification and properties of a novel raw starch degrading cyclomaltodextrin glucanotransferase from Bacillus firmus

  • B. N. Gawande
  • A. Goel
  • A. Y. Patkar
  • S. N. Nene
ORIGINAL PAPER

DOI: 10.1007/s002530051424

Cite this article as:
Gawande, B., Goel, A., Patkar, A. et al. Appl Microbiol Biotechnol (1999) 51: 504. doi:10.1007/s002530051424

Abstract

A novel raw starch degrading cyclomaltodextrin glucanotransferase (CGTase; E.C. 2.4.1.19), produced by Bacillus firmus, was purified to homogeneity by ultrafiltration, affinity and gel filtration chromatography. The molecular weight of the pure protein was estimated to be 78 000 and 82 000 Da, by SDS-PAGE and gel filtration, respectively. The pure enzyme had a pH optimum in the range 5.5–8.5. It was stable over the pH range 7–11 at 10 °C, and at pH 7.0 at 60 °C. The optimum temperature for enzyme activity was 65 °C. In the absence of substrate, the enzyme rapidly lost its activity above 30 °C. Km and kcat for the pure enzyme were 1.21 mg/ml and 145.17 μM/mg per minute respectively, with soluble starch as the substrate. For cyclodextrin production, tapioca starch was the best substrate used when gelatinized, while wheat starch was the best substrate used when raw. This CGTase could degrade raw wheat starch very efficiently; up to 50% conversion to cyclodextrins was obtained from 150 g/l starch without using any additives. The enzyme produced α-, β- and γ-cyclodextrins in the ratio of 0.2:9.2:0.6 and 0.2:8.6:1.2 from gelatinized tapioca starch and raw wheat starch with 150 g/l concentration respectively, after 18 h incubation.

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • B. N. Gawande
    • 1
  • A. Goel
    • 1
  • A. Y. Patkar
    • 1
  • S. N. Nene
    • 1
  1. 1.Chemical Engineering Division, National Chemical Laboratory, Pune 411 008, India e-mail: nene@ems.ncl.res.in Fax: +91-20-39 3041IN