Virgibacillus salarius BM02 was identified as a highly exopolysaccharide (EPS) producing bacterium. The EPS production
and its physico-chemical properties (intrinsic viscosity and total sugars/protein (TS/P) ratio) were optimized
using Box-Behnken experimental design. Maximum EPS production of 5.87 g L
−1
with TS/P ratio of 12.56
and intrinsic viscosity of 0.13 dL g
−1
was obtained at optimal conditions of sucrose (4.0% w/v), peptone (0.75% w/
v) and incubation period of 4.69 day. The monosaccharide composition of EPS was mannose, arabinose and glucose
at a molar ratio of 1.0:0.26:0.08. The EPS showed high water solubility (38.5%), water holding capacity
(514.46%) and foaming capacity (55.55%). The EPS showed moderate antioxidant activity in vitro and good emulsion
stabilizing properties against several hydrophobic compounds. The emulsifying activity was stable at different
temperatures, pH and ionic strength. Additionally, the acid hydrolysate of the EPS was evaluated as a carbon
source for the mixotrophic cultivation of industrially important Spirulina platensis. It induced an enhancement of
not only biomass production of S. platensis, but also cellular contents (pigments, proteins and lipids) leading to
higher nutritional value.

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The starting compound 4-amino-3-methyl-1-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide (1), that has been previously synthesized according to the literature procedure, underwent a reaction with the anhydride of phthalic acid either in AcOH or DMF to give isoindolinylpyrazole-5-carboxamide 3 and pyrazolothien-opyrimidoisoindoledione 4, respectively. Also, it was subjected to react with diethylmalonate followed by hydrazinolysis by hydrazine hydrate to yield the pyrazolothienopyrimidinyl acetohydrazide 5. The carbohydrazide derivative 5 was used as a key intermediate for the preparation of other new heterocyclic systems containing pyrazolylacetyl pyrazolothienopyrimidines and pyrazolothienopyrimidotriazepine compounds 6–11. The structures of these new heterocycles have been characterized by using analytical and spectroscopic analyses (IR, 1H-NMR, 13C-NMR and MS). Some derivatives of the synthesized compounds exhibited remarkable antibacterial and antifungal activities against many bacterial and fungal strains.

The starting compound 4-amino-3-methyl-1-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide (1), that has been previously synthesized according to the literature procedure, underwent a reaction with the anhydride of phthalic acid either in AcOH or DMF to give isoindolinylpyrazole-5-carboxamide 3 and pyrazolothien-opyrimidoisoindoledione 4, respectively. Also, it was subjected to react with diethylmalonate followed by hydrazinolysis by hydrazine hydrate to yield the pyrazolothienopyrimidinyl acetohydrazide 5. The carbohydrazide derivative 5 was used as a key intermediate for the preparation of other new heterocyclic systems containing pyrazolylacetyl pyrazolothienopyrimidines and pyrazolothienopyrimidotriazepine compounds 6–11. The structures of these new heterocycles have been characterized by using analytical and spectroscopic analyses (IR, 1H-NMR, 13C-NMR and MS). Some derivatives of the synthesized compounds exhibited remarkable antibacterial and antifungal activities against many bacterial and fungal strains.