Summary
A pullulan-hydrolysing enzyme of Micrococcus sp. 207 was purified to an electrophoretically homogenous state by chromatography on DEAE-Toyopearl, α-cyclodextrin-Sepharose and Asahipak GS-520P. The purified enzyme was free of α-amylase activity. The molecular weight of the enzyme as estimated by SDS-PAGE was 120,000 and the pI value as determined by isoelectric focusing was 4.9. The enzyme was most active at pH 8.0 and 50°C. The enzyme was activated by the addition of CaCl2, but its thermoresistance increased after removing free Ca2+ ions. The enzyme could hydrolyse the α-1,6-linkages of amylopectins, glycogens and pullulan and the K m value for pullulan was about 0.018%. Pullulan at concentrations above 0.012% inhibited the enzyme activity and the activity was competitively inhibited by cyclodextrins.
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Kimura, T., Horikoshi, K. Characterization of pullulan-hydrolysing enzyme from an alkalopsychrotrophic Micrococcus sp.. Appl Microbiol Biotechnol 34, 52–56 (1990). https://doi.org/10.1007/BF00170923
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DOI: https://doi.org/10.1007/BF00170923