Complementation of Sulfolobus solfataricus PBL2025 with an α-mannosidase: effects on surface attachment and biofilm formation

• Published in: Extremophiles : life under extreme conditions Vol. 16; no. 1; pp. 115 - 125
• Main Authors: Koerdt, A., Jachlewski, S., Ghosh, A., Wingender, J., Siebers, B., Albers, S.-V.
• Format: Journal Article
• Language: English
• Published: Japan Springer Japan 2012; Springer
• Subjects: alpha -Mannosidase; alpha-Mannosidase - metabolism; Animal, plant and microbial ecology; Base Sequence; beta -Galactosidase; Biochemistry; Biofilms; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Carbohydrates; Complementation; Confocal microscopy; DNA Primers; Enzymes; Fundamental and applied biological sciences. Psychology; Glucose; Life Sciences; Metabolism; Microbial Ecology; Microbiology; Microscopy, Confocal; Microscopy, Fluorescence; Original Paper; Polymerase Chain Reaction; Polysaccharides; Space life sciences; Sugar; Sulfolobus solfataricus; Sulfolobus solfataricus - enzymology; Sulfolobus solfataricus - metabolism; Surface Properties; Trees; Extracellular polymeric substances (EPS); Archaea; α-Mannosidase; Biofilm; Glycosylation; Lectin staining; Surface attachment; Lectin; Archaeobacteria; Enzyme; Staining; Sulfolobus solfataricus; Extracellular; Glycosylases; Sulfolobaceae; Glycosidases; Bacteria; Hydrolases; Complementation; Sulfolobales; Extreme environment
• ISSN: 1431-0651; 1433-4909
• DOI: 10.1007/s00792-011-0411-2

Compared to Sulfolobus solfataricus P2, the S. solfataricus mutant PBL2025 misses 50 genes (SSO3004-3050), including genes coding for a multitude of enzymes possibly involved in sugar degradation or metabolism. We complemented PBL2025 with two of the missing proteins, the α-mannosidase (SSO3006, Ssα-man) and the β-galactosidase LacS (SSO3019), and performed comparative fluorescence microscopy and confocal laser scanning microscopy to analyze the recombinant strains. We demonstrated that the Ssα-man complemented strain resembled the S. solfataricus P2 behavior with respect to attachment of cells to glass and growth of cells in static biofilms. During expression of the Ssα-man, but not LacS, glucose and mannose-containing extracellular polymeric substance (EPS) levels changed in the recombinant strain during surface attachment and biofilm formation. These results suggest that the Ssα-man might be involved in the modulation of the EPS composition and/or in the de-mannosylation of the glycan tree, which is attached to extracellular glycosylated proteins in S. solfataricus . On the other hand, LacS expression in PBL2025 reduced the carbohydrate content of the isolated total EPS implying a role in the modulation of the produced EPS during static biofilm formation. These are the first enzymes identified as playing a role in archaeal EPS formation.