Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110

Schaffert L, Schneiker-Bekel S, Gierhake J, Droste J, Persicke M, Rosen W, Pühler A, Kalinowski J (2020)
Applied Microbiology and Biotechnology 104: 5395–5408.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
Actinoplanes sp. SE50/110 (ATCC 31044) is the wild type of industrial producer strains of acarbose. Acarbose has been used since the early 1990s as an inhibitor of intestinal human α-glucosidases in the medical treatment of type II diabetes mellitus. The small secreted protein Cgt, which consists of a single carbohydrate-binding module (CBM) 20-domain, was found to be highly expressed in Actinoplanes sp. SE50/110 in previous studies, but neither its function nor a possible role in the acarbose formation was explored, yet. Here, we demonstrated the starch-binding function of the Cgt protein in a binding assay. Transcription analysis showed that the cgt gene was strongly repressed in the presence of glucose or lactose. Due to this and its high abundance in the extracellular proteome of Actinoplanes, a functional role within the sugar metabolism or in the environmental stress protection was assumed. However, the gene deletion mutant ∆cgt, constructed by CRISPR/Cas9 technology, displayed no apparent phenotype in screening experiments testing for pH and osmolality stress, limited carbon source starch, and the excess of seven different sugars in liquid culture and further 97 carbon sources in the Omnilog Phenotypic Microarray System of Biolog. Therefore, a protective function as a surface protein or a function within the retainment and the utilization of carbon sources could not be experimentally validated. Remarkably, enhanced production of acarbose was determined yielding into 8–16% higher product titers when grown in maltose-containing medium.
Stichworte
Biotechnology; Applied Microbiology and Biotechnology; General Medicine
Erscheinungsjahr
2020
Zeitschriftentitel
Applied Microbiology and Biotechnology
Band
104
Seite(n)
5395–5408
ISSN
0175-7598
eISSN
1432-0614
Page URI
https://pub.uni-bielefeld.de/record/2942996

Zitieren

Schaffert L, Schneiker-Bekel S, Gierhake J, et al. Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110. Applied Microbiology and Biotechnology. 2020;104:5395–5408.
Schaffert, L., Schneiker-Bekel, S., Gierhake, J., Droste, J., Persicke, M., Rosen, W., Pühler, A., et al. (2020). Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110. Applied Microbiology and Biotechnology, 104, 5395–5408. doi:10.1007/s00253-020-10584-1
Schaffert, L., Schneiker-Bekel, S., Gierhake, J., Droste, J., Persicke, M., Rosen, W., Pühler, A., and Kalinowski, J. (2020). Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110. Applied Microbiology and Biotechnology 104, 5395–5408.
Schaffert, L., et al., 2020. Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110. Applied Microbiology and Biotechnology, 104, p 5395–5408.
L. Schaffert, et al., “Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110”, Applied Microbiology and Biotechnology, vol. 104, 2020, pp. 5395–5408.
Schaffert, L., Schneiker-Bekel, S., Gierhake, J., Droste, J., Persicke, M., Rosen, W., Pühler, A., Kalinowski, J.: Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110. Applied Microbiology and Biotechnology. 104, 5395–5408 (2020).
Schaffert, Lena, Schneiker-Bekel, Susanne, Gierhake, Jessica, Droste, Julian, Persicke, Marcus, Rosen, Winfried, Pühler, Alfred, and Kalinowski, Jörn. “Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110”. Applied Microbiology and Biotechnology 104 (2020): 5395–5408.
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