Recent advances in glycobiotechnology show that bacterial exopolysaccharides (EPS) presenting glycosaminoglycan (GAG)-like properties can provide a valuable source of bio-active macromolecules for industrial applications. The HE800 EPS, named diabolican, is a marine-derived anionic high-molecular-weight polysaccharide produced by Vibrio diabolicus CNCM I-1629 which displays original structural features close to those of hyaluronic acid. We investigated the impact of carbon and nitrogen substrates on both Vibrio diabolicus growth and diabolican production. Both substrates were screened by a one-factor-at-a-time method, and experimental designs were used to study the effect of glucose, mannitol, and ammonium acetate various concentrations. Results showed that the medium composition affected not only the bacterium growth and EPS yield, but also the EPS molecular weight (MW). EPS yields of 563 and 330 mg L-1 were obtained in the presence of 69.3 g L-1 glucose and 24.6 g L-1 mannitol, respectively, both for 116.6 mM ammonium acetate. MW was the highest, with 69.3 g L-1 glucose and 101.9 mM ammonium acetate (2.3 × 106 g mol-1). In parallel, the bacterial maximum specific growth rate was higher when both carbon and nitrogen substrate concentrations were low. This work paves the way for the optimization of marine exopolysaccharide production of great interest in the fields of human health and cosmetics.

exopolysaccharide, production, molecular weight, Marine bacterium, central composite design, Vibrio diabolicus, yield

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]O[C@H]1[C@H](O)[C@@H](CO)O[C@@H](O[C@@H]2[C@@H](C(=O)O)O[C@@H](O[C@@H]3[C@@H](C(=O)O)O[C@@H](O[C@H]4[C@@H](CO)O[C@H]([*])[C@H](NC(C)=O)[C@H]4O)[C@H](O)[C@H]3O)[C@H](O)[C@H]2O)[C@@H]1NC(C)=O

758.636 g/mol (C₂₈H₄₂R₂N₂O₂₂, R = next and previous repeats, not counted in mol weight)

In the present study, a cost-effective, robust Microbioreactor based production optimization of levan like exopolysaccharide from marine Bacillus sp. SGD-03 was analysed. FE-SEM analysis has showed the significant fibrillar structure of EPS. Size exclusion chromatography and other analytical data revealed that, produced EPS has a molecular weight of 1.0 × 104 Da and is composed of fructose monosaccharide with hydroxyl, carbonyl, and ether groups. The backbone structure of EPS has a branching pattern of β-(2,6) linkages which confirms the similarity with available levan like polymers. The cost-effective media composition for levan production was demonstrated. The maximum yield of crude levan obtained was 123.9 g/L by response surface methodology using robust BioLector Pro Microbioreactor, and same has been validated with shake flask, 1 L and 10 L pilot-scale fermentation.

structure, EPS, marine, optimization, microbioreactor

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
	bDFruf	59.9	104.2	76.3	75.2	80.3	63.4


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
	bDFruf	3.60-3.71	-	4.12	4.03	3.89	3.48-3.83


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
	bDFruf	59.9/3.60-3.71		76.3/4.12	75.2/4.03	80.3/3.89	63.4/3.48-3.83

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]OC[C@H]1O[C@]([*])(CO)[C@@H](O)[C@@H]1O

162.141 g/mol (C₆H₁₀R₂O₅, R = next and previous repeats, not counted in mol weight)

A co-metabolization of xylose and glucose by Schizophyllum commune 227E.32 wild mushroom for exopolysaccharide (EPS) production is presented. Cultivations performed with S. commune 227E.32 at different xylose concentrations demonstrated that the concentration of 50 g·L-1 of xylose achieved the highest EPS production, around 4.46 g·L-1. Scale-up in a stirred tank reactor (STR) was performed. 10 % inoculum showed the highest cost/benefit ratio regarding sugar conversion and EPS production (Y P/S = 0.90 g·g-1), achieving 1.82 g·L-1 of EPS. Isolation, purification, and characterization were conducted with EPS produced in flasks and STR. GC-MS analysis showed glucose as main monosaccharide constituents for both isolates. 13C NMR and HSQC-edited showed that both EPS isolated consisted of a β-D-Glcp (1→3) main chain, partially substituted at O-6 with nonreducing β-D-Glcp ends on every third residue, similar to β-D-glucan isolated from S. commune basidiomes known as schizophyllan (SPG). The Mw was determined by GPC to 1.5 × 106 Da (flasks) and 1.1 × 106 Da (STR). AFM topographs revealed a semi-flexible appearance of the β-D-glucan, consistent with the triple helical structures adopted by SPG and overall contour length consistent with a high molar mass.

physicochemical characterization, mushroom β-(1 → 3)(1 → 6)glucan, xylose/glucose co-metabolization

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]O[C@H]1[C@H](O)[C@@H](CO)O[C@@H](O[C@@H]2[C@@H](O)[C@H](O[C@@H]3[C@@H](O)[C@H]([*])O[C@H](CO)[C@H]3O)O[C@H](CO[C@@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@H]3O)[C@H]2O)[C@@H]1O

648.564 g/mol (C₂₄H₄₀R₂O₂₀, R = next and previous repeats, not counted in mol weight)