Download Laboratory of microbial glycobiotechnology

Institute of Chemistry of SAS, Bratislava
Department of glycobiotechnology
Head of department: Dr Ján Tkáč, DSc.
Deputy of head of department: Dr. Desana Lišková, PhD.
Laboratory of microbial glycobiotechnology (Dr. Peter Gemeiner, DSc.):
1. Immobilization techniques for enhancement of stability of recombinant microbial cells and enzymes:
- as biocatalysts for one-step and cascade reactions for production of high added value chemicals
- advanced characterization techniques for immobilized viable whole cells
2. Biosensors as detectors for continuous monitoring and control of fermentation processes,
3. Lectins of plant origin applied in glycomics for prognosis of early stage pathological changes
Laboratory of bionanotechnology (Dr. Ján Tkáč, DSc.):
1. Construction of lectin biosensors modified with nanomaterials
2. Preparation of glycan biosensors with surface patterning conrolled at nanoscale
3. Application of nanomaterials for glycan enrichement
Laboratory of plant glycobiotechnology (Dr. Desana Lišková, PhD.):
1.Defense responses and protection of plants
2. Phytoremediations
3. Regulation of plant growth and differentiation
Financial support by the grants (2012-2016):
Slovak Grant Agency for Science VEGA: 7 projects
Science and Technology Assistance Agency APVV: 8 project
ITN scheme: 2 project
ERC grant: 1 project
Structural Funds of EU: 3 projects
Quatar National Research Fund: 1 project
COST Actions: 3 project
The main features of encapsulated and entrapped viable whole cells
as biocatalysts
PEC capsule
semipermeable membrane
B – biocatalyst
(cells, enzymes)
MWCO – molecular weight cut-off
entities with Mw > MWCO
encapsulated biocatalysts
The main purpose
Development of an universal immobilization technique
for any form of biocatalyst
benefits:
- separation, reusability
- continuous processes
- stabilization: operational, storage, temperature
- mechanical protection
Immobilization of viable recombinant E. coli with Baeyer-Villiger
monooxygenases in polyelectrolyte capsules and particles
• production of PEC microcapsules
(< 1mm)
Bučko et al. Appl Microbiol Biotechnol 100 (2016), 6585-6599
Techniques for preparation of immobilized viable cells
at the Department of glycobiotechnology
controlled production of immobilizates with wide range of parameters to set up
rate of flow, size of particles, uniformity, viscosity of hydrogel, sterility
Cutting of the flow
JetCutter®
p
Vibration system
EncapsBioSystems®
f [Hz]
p
Coaxial air jet
p
rpm
air
The concept of immobilized viable whole cell biocatalysts
for cascade reactions
substrate
A
Immobilized cells
in bioreactor
B
OH
E1
C
O
E2
D
O
E3
O
O
Biomass
E. coli cell factory
byproducts
and waste
Recycling
Product
How does the biosensor work?
Schematic principle of the work of biosensor
Open Chem., 2015; 13: 636–655
Amperometric biosensor
Calorimetric biosensor
(enzymatic thermistor)
Lectins 1
Lectins in glycobiology and diagnostic.
Lectins are the compounds having the protein or glycoprotein character. They
were found in plants, animals and in microbial organisms. Lectins have a potential
to act as biospecific recognition molecules of saccharides localized inside and
also on the surface of cells and in physiological liquids.
Lectin-saccharide interaction on the cell surface
Fluorescent lectin biochips: web page of the company MicroBioChips, Clin. Lab. Med. 2009
Med. Res. Rev. DOI 10.1002/med.21420
Plant cell wall and the biotechnologies
Plant cell wall is one of the most important protection mechanisms in plants
against environmental pollution and simultaneously also the source of
polysaccharide fragments – oligosaccharides, characterized by biological activity
with regulation effects having a large scale of utilization. The research on plant cell
wall at the Institute of Chemistry SAS is focused on the elucidation of the
relationship between the cell wall structure and function in defence and
regulation processes in plants.
Phytoremediation
Biotechnological method – phytoremediation - is utilizing plants with the
ability to accumulate e.g. toxic metals for environmental decontamination. Plant
cell walls serve as deposition place of toxic metals, as one of the plant defence
mechanisms. Cell wall derived-galactoglucomannan oligosaccharides (GGMOs),
nontoxic compounds for the environment, can increase the biomass and
tolerance/resistance of plant species against toxic metals and in such way
contribute to higher accumulation of the metal in plants used for
phytoremediation.
Regulation of plant growth and development
Regulation abilities of plant cell wall derived GGMOs consist in regulation of
growth (impact on seedlings root system, biomass formation in aboveground plant
parts), differentiation, regeneration and vegetative propagation of important
agricultural plants in conditions in vitro.
Impact of cadmium cations on cell wall structure
Changes induced by the presence of cadmium cations in cell wall
components proportion in maize
Protective effect of GGMOs against cadmium
 GGMOs reduce the negative effect of Cd
cations on primary root elongation and
structural changes in root apex.
Results obtained support the assumption
that GGMOs have protective effect
against abiotic stress.
Chronological distribution
of Cd cations in various plant
tissues
1st day – Cd is
predominantly
present in the
primary root
Primary root structure
suberin lamella
suberin lamella
xylem
xylem
3rd day – Cd is
present in the
primary root
and stem
suberin lamella
xylem
root hair
root hair
elongation
zone
root hair
elongation
zone
elongation
zone
meristem
meristem
meristem
control
cadmium
GGMOs + cadmium
5th day – Cd is present
in the primary root,
stem and leaves (also
in trichomes)
Impact of GGMOs on primary root growth and structure
mung bean
control
rhizodermis
GGMOs
rhizodermis
cortex
cortex
cell production rate
The primary
root elongation
zone is
obviously the
most important
site of GGMOs
action.
Root structure
altered
by altered
Root structure
in Arabidopsis
plants
by
galactoglucomannan
oligosaccharides
galactoglucomannan oligosaccharides
suberin lamella
xylem
suberin lamella
xylem
root
hair
root hair
elongation
zone
elongation
zone
Impact of GGMOs on plant cell
differentiation
meristem
80
Tracheary elements (%)
70
60
50
40
30
20
10
0
3
4
5
6
7
8
9
10
Days
GGMOs increased
formation of vessels tracheary elements
mediating transport of
solutions from roots to
aboveground plant parts.
control
meristem
GGMOs