title: Complementation of Sulfolobus solfataricus PBL2025 with an α-mannosidase: effects on surface attachment and biofilm formation.
abstract: 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.
authors.affiliation: Molecular Biology of Archaea, Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Strasse, Marburg, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Koerdt
authors.forename: S
authors.initials: S
authors.lastname: Jachlewski
authors.forename: A
authors.initials: A
authors.lastname: Ghosh
authors.forename: J
authors.initials: J
authors.lastname: Wingender
authors.forename: B
authors.initials: B
authors.lastname: Siebers
authors.forename: S-V
authors.initials: SV
authors.lastname: Albers
mesh_terms: D001483:Base Sequence; D018441:Biofilms; D017931:DNA Primers; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D048229:Sulfolobus solfataricus / Q000201:enzymology / Q000378:metabolism*; D013499:Surface Properties; D043323:alpha-Mannosidase / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D043323:alpha-Mannosidase
references.citation: Appl Environ Microbiol. 1996 Dec;62(12):4478-85
references.pmid: 16535464
references.citation: Appl Environ Microbiol. 2006 Jan;72(1):102-11
references.pmid: 16391031
references.citation: Biochimie. 2010 Dec;92(12):1895-907
references.pmid: 20696204
references.citation: Appl Environ Microbiol. 1988 Oct;54(10):2381-6
references.pmid: 16347749
references.citation: Arch Biochem Biophys. 2003 Jul 1;415(1):87-93
references.pmid: 12801516
references.citation: J Biol Chem. 2004 Feb 13;279(7):5537-48
references.pmid: 14645248
references.citation: Biotechnol Bioeng. 2000 Sep 5;69(5):537-47
references.pmid: 10898863
references.citation: Carbohydr Res. 1994 Jan 3;251:89-98
references.pmid: 8149382
references.citation: Biochim Biophys Acta. 1999 Dec 6;1473(1):96-107
references.pmid: 10580131
references.citation: J Biol Chem. 2005 Apr 29;280(17):16586-93
references.pmid: 15723834
references.citation: FEMS Microbiol Lett. 2004 Mar 12;232(1):1-6
references.pmid: 15061140
references.citation: Extremophiles. 2009 Jan;13(1):67-79
references.pmid: 18972064
references.citation: Methods Enzymol. 2001;330:201-15
references.pmid: 11210499
references.citation: Carbohydr Res. 1998 Jun;309(1):89-94
references.pmid: 9720239
references.citation: Can J Microbiol. 1995 Jan;41(1):12-8
references.pmid: 7728652
references.citation: Gene. 1990 Sep 28;94(1):89-94
references.pmid: 2121622
references.citation: PLoS One. 2010 Nov 24;5(11):e14104
references.pmid: 21124788
references.citation: J Biol Chem. 1976 Apr 10;251(7):2005-14
references.pmid: 1270419
references.citation: Mol Microbiol. 2003 Jun;48(5):1241-52
references.pmid: 12787352
references.citation: Extremophiles. 2010 Nov;14(6):485-91
references.pmid: 20835877
references.citation: Anal Biochem. 1977 Dec;83(2):346-56
references.pmid: 603028
references.citation: J Bacteriol. 2010 Jan;192(1):104-10
references.pmid: 19854908
references.citation: J Bacteriol. 1990 Dec;172(12):7111-8
references.pmid: 2123862
references.citation: Proc Natl Acad Sci U S A. 2011 Feb 22;108(8):3228-33
references.pmid: 21292982
references.citation: Carbohydr Res. 2009 Mar 31;344(5):648-53
references.pmid: 19203750
references.citation: Nat Rev Microbiol. 2010 Sep;8(9):623-33
references.pmid: 20676145
references.citation: J Biol Chem. 2007 May 11;282(19):14598-607
references.pmid: 17355973
references.citation: Arch Microbiol. 2008 Sep;190(3):371-7
references.pmid: 18438643
references.citation: Eur J Biochem. 2000 Jul;267(13):4144-9
references.pmid: 10866817
references.citation: J Ind Microbiol. 1996 Jun;16(6):331-41
references.pmid: 8987490
references.citation: Nat Rev Microbiol. 2011 Jun;9(6):414-26
references.pmid: 21572458
references.citation: Archaea. 2010 Sep 29;2010:
references.pmid: 20936123
references.citation: J Mol Biol. 2007 Dec 14;374(5):1224-36
references.pmid: 17996897
references.citation: J Bacteriol. 1999 Jul;181(14):4285-91
references.pmid: 10400586
references.citation: Arch Mikrobiol. 1972;84(1):54-68
references.pmid: 4559703
references.citation: J Bacteriol. 2003 Jul;185(13):3948-57
references.pmid: 12813089
references.citation: Mol Microbiol. 2007 May;64(3):795-806
references.pmid: 17462024
references.citation: Can J Microbiol. 2001 Jul;47(7):680-4
references.pmid: 11547890
references.citation: J Bacteriol. 2004 Jan;186(2):427-37
references.pmid: 14702312