Ebru Toksoy ner IBSB Industrial Biotechnology and Systems Biology - - PowerPoint PPT Presentation

IBSB – Industrial Biotechnology and Systems Biology Research Group

Workshop BioGlue

Vienna – Nov 2010

Marmara University, Department of Bioengineering, Istanbul, Turkey

Ebru Toksoy Öner

• IBSB
• Extremophiles
• Levan by Bacillus sp.
• Extremophilic Levan

Industrial Biotechnology and Systems Biology Research Group

Marmara University Department of Bioengineering Istanbul, Turkey

http://ibsb. marmara.edu.tr

Ebru Toksoy Oner, Assoc. Prof. Kazım Yalcın Arga, Assist. Prof. 1 Post-Doc 8 pHD students 2 MS students

Industrial Biotechnology and Systems Biology Research Group

Marmara University Department of Bioengineering Istanbul, Turkey

http://ibsb. marmara.edu.tr

Betül Kırdar, Prof. Barbara Nicolaus, Prof. Stephen G. Oliver, Prof. Robert Dekker, Prof.

• interpret and optimize production capabilities
• f yeast, bacteria and extremophiles via systems

based approach

• optimization of fermentation processes to

design high-yield production lines

• production of biopolymers form extremophiles
• isolation and identification of extremophiles
• strain improvement for bioethanol production

Ion N. Mihailescu, Prof.

Extreme conditions can refer to physical extremes (temperature, pressure

• r

radiation)

• r

geochemical extremes (salinity, desiccation,

• xygen

tension and pH). Extremophiles are microorganisms that no not

• n
• nly

ly tolerate

• lerate

such extreme conditions, bu but usually ly re requ quire ire such environmental extremes for their survival and growth. ecological systems such as hot springs, salt and soda lakes, deserts and ocean beds that are not compatible with human life are considered as being extreme.

Anoxybacillus amylolyticus MR3CT Isolated from M. Rittmann (Antarctica) T 60°C Geobacillus thermoleovorans subsp. stromboliensis sbsp.nov Isolated from the geothermal volcanic environment (Italy) T 70°C Halomonas sp. AAD6 Isolated from Camaltı Saltern Area (Turkey)

Starchy agro- industrial wastes Bioethanol…

to success essfully fully enginee eer Biopoly lymer propertie ies and improv

• ve

e productio ion

System ems Biolog logy Aprroa

• ach

Protein-encoding gene models 584 Metabolites 1389 Intracellular metabolites 1020 Extracellular metabolites 369 Reactions 1080 Enzymatic reactions 870 Transport fluxes 210

Metabol

• lic Model of Halomonas

s sp.

Micr crobial l Produ duct ctio ion

Nanostructured Coatings for Biomedical Applications

 Thin, high-quality and uniform films

produced by the MAPLE (Matrix Assisted Pulsed Laser Evaporation) technique

Micr crobial l Produ duct ctio ion

Chemical media Low cost substrates  Levan – based micro/nano particles

may be a potential delivery system for macromolecules.

Drug Delivery Systems Environmental Applications

 Emulsifying Agent Bioflocculating activity

Functional Biofilms

 Edible Food Packaging

FRUCTAN



a homopolimer of fructose units (polyfructose, fructan)

 a crucial component of drought and freeze protection in plants 

ability to stabilize membranes in dry and cold environments

Levan Inulin Graminan

complex and branched linear β-(2,6)-linked fructofuranosyl units linear β-(2,1)-linked fructofuranosyl units

• Blood plasma extender
• Hypocholesterolaemic agent
• Anti-AIDS agent
• Immunostimulating agent
• Tablet binder
• Add sweetness to foods
• prebiotics
• Filler
• Bulking agent
• Substitute for gums
• Animal feed
• Edible food packaging
• Carrier for flavor and

fragrances

• bioadhesive
• Provide viscosity
• Holding capacity for water

and chemicals

• Selective plugging agent
• Source for pure fructose
• cryoprotectant
• Emulsifier
• Formulation aid
• Stabilizer and thickener
• Surface-finishing agent
• Encapsulating agent
• Cosmetic
• ….

Medica cal Pharma rmaceut ceutica cal Others ers Indust strial ial Fo Foods

BENEF EFITS S : Produced from renewable resource, sugar. No petroleum or natural gas derivatives in product. No VOCs, HAPs or other toxic emissions. Water-based with no solvents. No health issues for users. No dermal irritation. No allergic contact sensitization. Biodegradable. Reduce regulatory burden. Long term storage as powder.

 produced by Bacillus sp. has strong bioadhesive properties  hydroxyl groups in its structure form adhesive bonds with various substrates. the most promising commercial polysaccharide based adhesives are actually made from levan

• Combie, J. (2005) Properties of Levan and Potential Medical Uses, Polysaccharides for Drug Delivery and PharmaceuticalApplications, June 22, 2006. Vol.934, 263-269.
• Mancuso Nichols et al. (2009) 'Screening Microalgal Cultures in Search of Microbial Exopolysaccharides with Potential as Adhesives', The Journal of Adhesion, 85: 2, 97 — 125.

http://specialtybiopolymers.com

• Dr. Joan Combie

Montana Polysaccharides Corp. (USA)

Natural Polymer Tensile Strength psi Levan 991 Carboxymethylcellulose 193 Inulin 124 Guar gum 63 Xanthan gum 33

 Very limited information and literature  Not produced at large scale  Production conditions depend on the microbial system used  MW and degree of branching depend on the production conditions  Biological activity and physicochemical properties depend on production  Strict control over process parameters is necessary  Difficult to purify  Expensive

an optimal cost-effective production process is a must!

Plant

• Grasses (Dactylis glomerata, Poa secunda, Agropyron cristatum)

Microorganisms

• Zymomonas mobilis
• Bacillus sp.
• Erwinia herbicola
• Gluconoacetobacter xylinus
• Microbacterium laevaniformans
• Rahnella aquatilis
• Serratia levanicum
• Pseudomonas syringae
• Halomonas sp.

* Poli A. et al.. 2009. Carbohydrate Polymers, 78, 651-657.

the first and only levan-producer extremophile * Highest production yield on available substrate

Microb

• bial

Production

• n by

Halomonas sp.

Chemical media + sucrose Sugar beet molasses

10 X increase in uronic acid content !!

Biocompatibility studies showed that levan produced by Halomonas sp. did not affect cellular viability and proliferation of osteoblasts and murine macrophages suggesting the high biocompatibility of this EPS.

Brine Shrimp Test. The inhibition of avarol toxic activity on brine shrimp (Artemia salina) test was performed in artificial sea water. With decreasing doses of levan solution (500, 50, 5 ppm), protective effect against the toxic activity of avarol increased. Halomonas levan was found to increase the LD50 value of avarol from 0.18 ppm up to 10

• ppm. The protective effect of the polymer against the toxic activity of

Avarol implied its potential use as an anti-cytotoxic agent. MTT cell proliferation assay was employed to assess the cell

• viability. Levan by Halomonas sp.

showed high biocompatibility and affinity against both cell lines.

cancerous cell line He La (human cervical cancer cells) non-cancerous cell line L929 (mouse fibroblast cells) mouse monocyte/macrophage cell line J774

• steoblast cells isolated from

the calvaria of Wistar rats

Idea dea:

extreme conditions (salinity) at which levan is microbially produced may also confer it some unique properties enhancing its adhesive strength !

current research efforts on levan from Halomonas sp. are now focused on elucidating its potential to be used as a commercially useful adhesive by



Developing new formulations with the polymer and its modified forms



Understanding its mechanism of action

http://ibsb. marmara.edu.tr

Thank you for your listening !

http://ibsb. marmara.edu.tr