Pullulan polysaccharide and its derivatives as bioplastic

M. Moscovici1, Corina Ionescu1, Ortansa Fotea1 Georgeta Mocanu2, Doina Mihai2, A.Carpov2
National Institute for Chemical-Pharmaceutical Research and Development, Bucharest, Romania 2) Petru Poni Institute of Macromolecular Chemistry, Iasi, Romania
1)

Bioreactor fermentation

Typical fermentation profiles of A. pullulans ICCF-83 cultivation: a) batch; b) fed-batch. Table 1: Pullulan biosynthesis by batch and fed-batch fermentation with A. pullulans ICCF-83 (best results) [6]. Bioreactor Cultivation technique Performances: Polysaccharide production Maximum final concentration, g/l Productivity, volumetric g/l.h Substrate (carbohydrate) conversion yield, %
* On an optimized medium composition

Microfermenter (14 l) batch fed-batch 50 - 74* 1.06 64 103 0.99 76

Pilot tank reactor (150 l) batch fed-batch 60 0.79 66 101 0.90 63

Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania

Downstream process flowsheet for isolation and purification of pullulan - polysaccharide.

Broth
Filter aid Biomass separation Centrifuging / Precoat filtration Wet biomass Disposal

Supernatant / Filtrate solution

Concentration Vacuum evaporation Water Disposal Mother liquor (methanol aqueous Methanol recovery

Concentrate
Methanol Crude product isolation Precipitation, centrifuging / filtration, drying

Crude pullulan
Water

Applications
Dissolving, filtration

Pullulan solution
Deionized water Concentration - purification Ultrafiltration - diafiltration Spent aqueous Disposal

Purified concentrate solution

Methanol Purified product isolation Precipitation, centrifuging / filtration, drying Mother liquor (methanol aqueous) Methanol recovery

Pullulan polysaccharide (purified)

Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania

Product Quality Analysis

Chromatographic tests concerning the purity of pullulan:

(a)

(b)

TLC: a, b - reference; c, d - ICCF sample; a, c - enzymatic hydrolysis (pullulanase); b, d - chemical hydrolysis; e - glucose.

HPLC: (a) - reference; (b)- ICCF sample

Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania

Reduced viscosity versus polymer concentration for pullulan, carboxymethyl pullulan (CMP) and its associative derivatives, substituted with different content of hydrophobic groups (o): pullulan; () CMP; ( ) CMP-1.3 C16; ( ) CMP-3.4 C16; ( ) CMP-4.5 C16; () CMP-6.8 C16.

Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania

Effect of the reaction temperature on the DS with chloroacetyl groups of dextran and pullulan
molar ratio chloroacetyl chloride/ glucopyranosic unit: 5/ 1; 6h, solvent: dimethyl formamide.

Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania

Influence of the temperature on the %Cl (DSCl) and %S (DSS) of the reaction products;
solvent: pyridine 6 h, molar ratios MsCl/glucosidic unit: () 1/1; (o) 2/ 1; (x) 3/ 1.

Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania

In vitro release curves of metronidazole from its conjugates with pullulan previously chloroacylated with various alkyl length
50 pH=1.6; C5 40 g% MI released pH=1.6; C4 pH=1.6; C3 pH=1.6; C2 20 pH=7.4; C5 pH=7.4; C4 10 pH=7.4; C3 pH=7.4; C2 0 4 8 12 T ime, h
Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania 8

30

0 16 20 24

Conclusions
Mutant high-producing strains of Aureobasidium pullulans Biosynthetic performances of pullulan polysaccharide fermentation: final polysaccharide concentration (g/l): batch: 50 - 74 fed-batch: 101 - 103 volumetric productivities (g/lh): 0.8 - 1.1 substrate conversion yield (%): 63 - 76 Downstream isolation and purification process - purified, narrow MW distributed product. Synthesis of new pullulan derivatives: cross-linked with reactive groups assocative (amphiphillic) polymers Good outlook for a further development of: a commercial process synthesis of new products enlarging the field of potential applications and knowledge
Pullulan polysaccharide and its derivatives as bioplastic M. Moscovici, G. Mocanu - Romania 9