English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Structural and functional characterization of three novel fungal amylases with enhanced stability and pH tolerance.

MPS-Authors
/persons/resource/persons213998

Roth,  Christian
Christian Roth, Biomolekulare Systeme, Max Planck Institute of Colloids and Interfaces, Max Planck Society;

External Resource
No external resources are shared
Fulltext (public)

Article.pdf
(Publisher version), 6MB

Supplementary Material (public)
There is no public supplementary material available
Citation

Roth, C., Moroz, O. V., Turkenburg, J. P., Blagova, E., Waterman, J., Ariza, A., et al. (2019). Structural and functional characterization of three novel fungal amylases with enhanced stability and pH tolerance. International Journal of Molecular Sciences, 20(19): 4902. doi:10.3390/ijms20194902.


Cite as: http://hdl.handle.net/21.11116/0000-0005-3E38-8
Abstract
Amylases are probably the best studied glycoside hydrolases and have a huge biotechnological value for industrial processes on starch. Multiple amylases from fungi and microbes are currently in use. Whereas bacterial amylases are well suited for many industrial processes due to their high stability, fungal amylases are recognized as safe and are preferred in the food industry, although they lack the pH tolerance and stability of their bacterial counterparts. Here, we describe three amylases, two of which have a broad pH spectrum extending to pH 8 and higher stability well suited for a broad set of industrial applications. These enzymes have the characteristic GH13 alpha-amylase fold with a central (beta/alpha)8-domain, an insertion domain with the canonical calcium binding site and a C-terminal beta-sandwich domain. The active site was identified based on the binding of the inhibitor acarbose in form of a transglycosylation product, in the amylases from Thamnidium elegans and Cordyceps farinosa. The three amylases have shortened loops flanking the nonreducing end of the substrate binding cleft, creating a more open crevice. Moreover, a potential novel binding site in the C-terminal domain of the Cordyceps enzyme was identified, which might be part of a starch interaction site. In addition, Cordyceps farinosa amylase presented a successful example of using the microseed matrix screening technique to significantly speed-up crystallization.