Improved Isolation of SlaA and SlaB S-layer proteins in Sulfolobus acidocaldarius

Simonin, P. A.; Lombard, Carine; Huguet, Arnaud; Kish, Adrienne

doi:10.1007/s00792-020-01179-9

Cited by 3 publications

(3 citation statements)

References 42 publications

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“…Although no signal peptide could be detected with SignalP 5.0 [53], the PRED-TAT server [54] predicted a clear signal peptide (reliability score: 0.875), as it also did for SlaA that served as positive control (reliability score 0.925). SlaA is the best characterized extracellular S-layer protein from the thermoacidophilic archaeon Sulfolobus acidocaldarius [55]. This predicted extracellular localization of HmcIsc is in full accordance with the previously reported purification of inulin-type fructans from the Suc supplemented growth medium of Hmc.…”

Section: Orf Analysis Of the Halomicrobium Sp Ibsba Gh68 Membersupporting

confidence: 87%

“…50 mM sodium acetate buffer for pH 4 and pH 5, 50 mM potassium phosphate buffer for pH 6 to pH 7.5, 50 mM Tris buffer for pH 8.0 and pH 8.5 and 50 mM carbonate-bicarbonate buffer for pH 9.0 were used. Reactions were carried out at 45 • C for 24 h. To analyze the effect of temperature, standard reactions were incubated at different temperatures (4,15,37,45,50,55,60,65, and 70 • C). Enzyme activities were determined with DNS method at 24 h.…”

Section: Effects Of Ph and Temperaturementioning

confidence: 99%

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Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase

et al. 2021

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Fructans are fructose-based (poly)saccharides with inulin and levan being the best-known ones. Thanks to their health-related benefits, inulin-type fructans have been under the focus of scientific and industrial communities, though mostly represented by plant-based inulins, and rarely by microbial ones. Recently, it was discovered that some extremely halophilic Archaea are also able to synthesize fructans. Here, we describe the first in-depth functional and molecular characterization of an Archaeal inulosucrase from Halomicrobium sp. IBSBa (HmcIsc). The HmcIsc enzyme was recombinantly expressed and purified in Escherichia coli and shown to synthesize inulin as proven by nuclear magnetic resonance (NMR) analysis. In accordance with the halophilic lifestyle of its native host, the enzyme showed maximum activity at very high NaCl concentrations (3.5 M), with specific adaptations for that purpose. Phylogenetic analyses suggested that Archaeal inulosucrases have been acquired from halophilic bacilli through horizontal gene transfer, with a HX(H/F)T motif evolving further into a HXHT motif, together with a unique D residue creating the onset of a specific alternative acceptor binding groove. This work uncovers a novel area in fructan research, highlighting unexplored aspects of life in hypersaline habitats, and raising questions about the general physiological relevance of inulosucrases and their products in nature.

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Section: Orf Analysis Of the Halomicrobium Sp Ibsba Gh68 Membersupporting

confidence: 87%

Section: Effects Of Ph and Temperaturementioning

confidence: 99%

Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase

et al. 2021

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“…of the S-layer is not completely explained already. However, in addition to its adhesion ability, various known functions of the S-layer include cell protection against environmental hazards, control of nutrient-metabolite transfer, aggregation, support in surface recognition [24], and protective role against digestive enzymes [20,21,24,25].…”

Section: Introductionmentioning

confidence: 99%

Strain‐dependent effectivity, and protective role against enzymes of S‐layers in Lactiplantibacillus plantarum strains

Alp

2022

J Basic Microbiol

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The present study investigated whether the surface layer (S-layer), which is known to have a varying effect from strain to strain on aggregation, adhesion ability, also has an effect on the resistance of bacteria to digestive enzymes, phenol, lysozymes. The effect of S-layers on the resistance against various enzymes, aggregation and adhesion abilities, and strain specificity were determined of eight Lactiplantibacillus plantarum strains. Strains were treated with 5 M lithium chloride (LiCl) to extract the S-layers, the presence of this layer in those microorganisms was demonstrated by polyacrylamide gel electrophoresis. Scanning electron microscopy was used to visualize the separation of the S-layer, which surrounds the microorganism, from the microorganism by the LiCl. The images were taken three times, once at the beginning, once 30 min later, and once at the end of this process, which took 2 h in total. The effect against enzymes varied depending on the strain, but it was determined that all the tested strains had a serious loss of viability against phenol in the absence of an S-layer. Lpb. plantarum DA100 showed a maximum decrease against gastrointestinal system enzymes after the LiCl (96.48 ± 0.03% before and 66.46 ± 0.01% after LiCl). Lpb. plantarum DA255 showed a significant decrease against lysozyme (99.11 ± 0.00% before and 62.80 ± 0.0% after LiCl). Removal of the S-layer greatly affected the adhesion ability of some strains, while for others there was hardly any change. The results showed that the role of the S-layer may be strain-specific, the rate of effect can vary. The primary function of S-layer proteins is thought to contribute to the adhesion ability of bacteria. There are limited studies that have reported the protective property of this layer against various enzymes, however, our results showed that S-layer could be one of the resistance strategies developed by bacteria against enzymes.

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A new and improved protocol for extraction of intact polar membrane lipids from archaea

Evans

Elling

et al. 2022

Organic Geochemistry

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Improved Isolation of SlaA and SlaB S-layer proteins in Sulfolobus acidocaldarius

Cited by 3 publications

References 42 publications

Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase

Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase

Strain‐dependent effectivity, and protective role against enzymes of S‐layers in Lactiplantibacillus plantarum strains

A new and improved protocol for extraction of intact polar membrane lipids from archaea

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