New type of pullulanase from Bacillus stearothermophilus and molecular cloning and expression of the gene in Bacillus subtilis
Open Access
- 1 April 1988
- journal article
- research article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 170 (4), 1554-1559
- https://doi.org/10.1128/jb.170.4.1554-1559.1988
Abstract
A new type of pullulanase which mainly produced panose from pullulan was found in Bacillus stearothermophilus and purified. The enzyme can hydrolyze pullulan efficiently and only hydrolyzes a small amount of starch. When pullulan was used as a substrate, the main product was panose and small amounts of glucose and maltose were simultaneously produced. By using pTB522 as a vector plasmid, the enzyme gene was cloned and expressed in Bacillus subtilis. Since the enzyme from the recombinant plasmid carrier could convert pullulan into not only panose but also glucose and maltose, we concluded that these reactions were due to the single enzyme. The new pullulanase, with a molecular weight of 62,000, was fairly thermostable. The optimum temperature was 60 to 65 degrees C, and about 90% of the enzyme activity was retained even after treatment at 60 degrees C for 60 min. The optimum pH for the enzyme was 6.0.This publication has 40 references indexed in Scilit:
- Entire nucleotide sequence of the pullulanase gene of Klebsiella aerogenes W70Journal of Bacteriology, 1987
- Effect of in vitro DNA Rearrangement in the NH2-terminal Region of the Penicillinase Gene from Bacillus licheniformis on the Mode of Expression in Bacillus subtilisMicrobiology, 1985
- MIXED CONNECTIVE TISSUE DISEASE WITH FATAL PULMONARY HYPERTENSIONActa Pathologica Japonica, 1982
- A Cultural Method for the Detection of Pullulan-Degrading Enzymes in Bacteria and its Application to the Genus BacillusJournal of Applied Bacteriology, 1979
- Hydrolysis of Pullulan by a Novel Enzyme fromAspergillus nigerAgricultural and Biological Chemistry, 1971
- Polysaccharide Produced by the Genus PullulariaApplied Microbiology, 1963
- The Hydrolysis of Glucosyl Oligosaccharides with α-d-(1→4) and α-d-(1→6) Bonds by Fungal AmyloglucosidaseJournal of Biological Chemistry, 1960
- Colorimetric Method for Determination of Sugars and Related SubstancesAnalytical Chemistry, 1956
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951
- Detection of Sugars on Paper ChromatogramsNature, 1950