Fluorescence Lectin Bar-Coding of Glycoconjugates in the Extracellular Matrix of Biofilm and Bioaggregate Forming Microorganisms
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microbial Strains and Culture Conditions
2.2. Lectin-Screening
2.3. Epifluorescence and Laser Microscopy
2.4. Binary Bar-Coding and Heat-Mapping
2.5. Digital Image Analysis
3. Results
3.1. Visual Examination
3.2. Binary Bar-Coding
3.3. Heat Map Bar-Coding
3.4. Lectin-Binding Information
4. Discussion
5. Conclusions and Perspective
- Fluorescence lectin bar-coding (FLBC) represents a powerful approach for selecting a panel of lectins useful for subsequent fluorescence lectin-binding analysis (FLBA).
- The time needed for lectin bar-coding is dependent on the difficulty in mounting the samples. It will also depend on the number of lectins bound, which is equal to the number of datasets to be recorded.
- Usually lectin bar-coding on one sample type with the current range of commercially available lectins (≈70) can be easily performed within two days.
- Subsequent fluorescence lectin-binding analysis (FLBA) with a selected panel of positive lectins is often combined with nucleic acid staining. This allows visualization of bacterial cell and glycoconjugate distribution as well as lectin-specific glycoconjugate typing see [16].
- The advantage of lectin bar-coding as compared to microarray techniques lies in the immediate visual information and identification of the lectin binding pattern.
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
DSM | Deutsche Sammlung für Mikroorganismen |
DSMZ | Deutsche Sammlung für Mikroorganismen und Zellkulturen |
ASW | artificial seawater |
TSB | tryptic soy broth |
THB | Todd Hewitt broth |
MAC | Mackintosh basal salt solution |
NYG | nutrient broth supplemented with yeast extract and glucose |
YEB | complex medium with yeast extract, beef extract, peptone and sucrose |
Fuc | Fucose |
Gal | Galactose |
GalNAc | N-Acetyl galactosamine |
Glc | Glucose |
GlcNAc | N-Acetyl glucosamine |
Man | Manose |
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Strain | Medium/Growth Conditions/Reference |
---|---|
Various diatoms | ASW plus inorganic nutrients, liquid culture |
Amphora caffeaeformis | [21] |
Craspedostauros australis | [21] |
Various cyanobacteria | BG 11, liquid culture |
Metallosphaera hakonensis | DSMZ 88 medium, 65 °C, pH 2.5, grown on 10 µm chalcopyrite grains in Erlenmeyer flasks |
Leptospirillum ferriphilum | MAC, 0.02 % yeast extract, pH 1.8, pyrite grains in Erlenmeyer flasks |
Acidianus sp. DSM 29099 | [20] |
Ferroplasma acidiphilum DSM 29986 | [20] |
Sulfolobus metallicus DSM 6482 | [20] |
Ferrovum C4 | iFeo medium, pH 2.5, liquid culture |
Deinococcus geothermalis | [16] |
Pseudomonas syringae | [15] |
Beggiatoa | [22] |
Bacillus cereus 10987 | TSB + 0.5% yeast extract, IBIDI 96 well microplates |
Streptococcus epidermidis 1457 | TSB, IBIDI 96 well microplates |
Campylobacter jejuni | [23] |
Sphingobium sp. Z007 | [24] |
Lactobacillus sp. | THB, liquid culture, 35 °C |
Streptococcus mitis | THB, liquid culture, 35 °C |
Acinetobacter naesludii | THB, liquid culture, 35 °C |
Burkholderia cenocepacia H111 | NYG, YEB, membrane filters on agar plates |
Lectin | Specificity |
---|---|
AAL | α-Fuc |
IAA | n. d. |
WGA | β-GlcNAc |
VVA | α-Man, α-GalNAc |
LEA | β-GlcNAc |
PNA | β-Gal |
AIA | n.d. |
ECA | α-Gal, β-Gal, α-GalNAc, β-GalNAc |
HAA | α-GlcNAc, α-GalNAc |
HPA | α-GalNAc |
PSA | α-Man, α-Glc, α-GlcNAc |
SBA | α-GalNAc, β-GalNAc |
WFA | α-GalNAc, β-GalNAc |
DBA | α-GalNAc |
LcH | α-Man, α-Glc, α-GlcNAc |
MNA | n.d. |
PHA-E | n.d. |
STA | β-GlcNAc |
UEA | β-GlcNAc, α-Fuc |
GS-I | α-Gal, α-GalNAc |
HHA | α-Man |
MPA | n.d. |
PMA | n.d. |
VGA | n.d. |
ConA | α-Man, α-Glc, α-GlcNAc |
GNA | α-Man |
HMA | α-Fuc, α-GalNAc |
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Neu, T.R.; Kuhlicke, U. Fluorescence Lectin Bar-Coding of Glycoconjugates in the Extracellular Matrix of Biofilm and Bioaggregate Forming Microorganisms. Microorganisms 2017, 5, 5. https://doi.org/10.3390/microorganisms5010005
Neu TR, Kuhlicke U. Fluorescence Lectin Bar-Coding of Glycoconjugates in the Extracellular Matrix of Biofilm and Bioaggregate Forming Microorganisms. Microorganisms. 2017; 5(1):5. https://doi.org/10.3390/microorganisms5010005
Chicago/Turabian StyleNeu, Thomas R., and Ute Kuhlicke. 2017. "Fluorescence Lectin Bar-Coding of Glycoconjugates in the Extracellular Matrix of Biofilm and Bioaggregate Forming Microorganisms" Microorganisms 5, no. 1: 5. https://doi.org/10.3390/microorganisms5010005