Cloning and Partial Characterization of an Endo-α-(1→6)-d-Mannanase Gene from Bacillus circulans
Abstract
:1. Introduction
2. Results and Discussion
2.1. Cloning of the Full-Size Endo-α-(1→6)-d-Mannanase Gene from Bacillus circulans TN-31
2.2. Expression and Purification of the Glycosyl Hydrolase Domain of Emn
2.3. Digestion of Synthetic Mannoside Substrates
2.4. Digestion of Phosphatidyl-Myo-Inositol-Mannosides
2.5. Analysis of Endomannanase-Digested LAM and LM
3. Materials and Methods
3.1. Cloning, Expression and Purification the Glycosyl Hydrolase Domain of Emn
3.2. Assays Using Synthetic Mannosides Substrates
3.3. Assays Using Purified Mycobacterial PIMs and Lipoglycans
3.4. Analysis of Substrates and Reaction Products by Liquid Chromatography—Mass Spectrometry
4. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Angala, S.k.; Li, W.; Palčeková, Z.; Zou, L.; Lowary, T.L.; McNeil, M.R.; Jackson, M. Cloning and Partial Characterization of an Endo-α-(1→6)-d-Mannanase Gene from Bacillus circulans. Int. J. Mol. Sci. 2019, 20, 6244. https://doi.org/10.3390/ijms20246244
Angala Sk, Li W, Palčeková Z, Zou L, Lowary TL, McNeil MR, Jackson M. Cloning and Partial Characterization of an Endo-α-(1→6)-d-Mannanase Gene from Bacillus circulans. International Journal of Molecular Sciences. 2019; 20(24):6244. https://doi.org/10.3390/ijms20246244
Chicago/Turabian StyleAngala, Shiva kumar, Wei Li, Zuzana Palčeková, Lu Zou, Todd L. Lowary, Michael R. McNeil, and Mary Jackson. 2019. "Cloning and Partial Characterization of an Endo-α-(1→6)-d-Mannanase Gene from Bacillus circulans" International Journal of Molecular Sciences 20, no. 24: 6244. https://doi.org/10.3390/ijms20246244