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A three-variable Box-Behnken design was employed to obtain the best possible combination of extraction time, ratio (raw material/water) and extraction temperature to allow maximum extraction yield of polysaccharides from pea pod (PPP). The preferred extraction conditions were: extraction time 195 min, extraction temperature 70 °C and ratio of raw material/water 1/40. Under these conditions, the experimental yield was 16.21 ± 1.12%, which is in close agreement with the value predicted by response surface methodology model yield (16.08 ± 0.95%). The molecular weight distribution of PPP showed two peaks with MW of 5217 kDa and 50 kDa, respectively. The main monosaccharides in PPP were galactose, xylose and arabinose, whereas the major functional groups identified from FT-IR spectrum included CO, OH and CH. In addition, PPP had high 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and a moderate reducing power. The antibacterial activity of PPP was also observed against the tested microorganisms and at 50 mg/mL PPP could inhibit the growth of the Gram-negative and Gram-positive bacteria. Generally, these results suggest that the PPP has significant antioxidant activity and good antibacterial activity and can potentially be used as additive in food, pharmaceutical and cosmetic preparations.

This work aims at realizing an optimal hydrolysis of pretreated Alfa fibers (Stipa tenacissima) through the use of enzymes produced from Talaromyces thermophilus AX4, namely β-d-glucosidase and xylanase, by a solid state fermentation process of an agro-industrial waste (wheat bran supplemented with lactose). The carbon source was firstly selected and the optimal values of three other parameters were determined: substrate loading (10g), moisture content (85%) and production time (10days); which led to an optimized enzymatic juice. The outcome was then supplemented with cellulases of T. reesei and used to optimize the enzymatic saccharification of alkali-pretreated Alfa fibers (PAF). The maximum saccharification yield of 83.23% was achieved under optimized conditions (substrate concentration 3.7% (w/v), time 144h and enzyme loading of 0.8 FPU, 15U CMCase, 60U β-d-glucosidase and 125U xylanase).The structural modification of PAF due to enzymatic saccharification was supported by the changes of morphologic and chemical composition observed through macroscopic representation, FTIR and X-Ray analysis.

The present study investigates the hypolipidemic effects of sulphated polysaccharide obtained from Codium fragile (CFSP) in induced obese rats (HFD). The results showed an increase in body weight of HFD rats by 21.56% as compared to control normal rats. Moreover, serum lipase activity underwent an increase which led to an increase in the levels of total cholesterol (T-Ch), triglycerides (TG) and low density lipoprotein cholesterol (LDL-Ch) in serum associeted with a decrease in the level of high density lipoprotein cholesterol (HDL-Ch) in untreated HFD rats. This diet has disrupted the antioxidant status by decreasing the activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX)) and subsequently an increase in thiobarbituric acid reactive substances (TBARS) level in liver and kidney of obese rats. All these disturbances are significantly corrected by CFSP administration with no fatty deposits in the liver and a protective effect against renal histological alteration. This confirms the important role of this polysaccharide in the fight against oxidative stress and the prevention of hyperlipidemia.

A newly profiling of Salicornia arabica (SAPS) polysaccharides was investigated. The water-soluble polysaccharide extract extracted from the Tunisian halophyte (SA) contained sulfate (9.64 ± 0.003%), carbohydrate (86.33 ± 0.003%), protein (1.24%), and uronic acids (3.33 ± 0.37%). The analysis of the monosaccharide composition revealed the presence of arabinose, Glucopyranose galactose, Glucuronic acid and glucose. The identified polysaccharide was considered as heterogeneous and highly branched via gaz chromatography, Fourier Transform Infrared Spectrometer (FT-IR), Thin layer chromatography (TLC) analysis. It also displayed an important radical scavenging and reducer power as well it exhibited effective antimicrobial properties.

A newly isolated bacterial strain from Tunisian thermal source was selected for its ability to produce extracellular levanase when grown on levan substrate. The optimization of carbon source, nitrogen source, temperature and initial pH of the growth medium in submerged liquid cultures were investigated. In fact, levan was found to be a good inducer of levanase enzymes. The optimal temperature and pH of the levanase activity were 40 °C and 6.4, respectively. This enzyme exhibited a remarkable stability and retained 75% of its original activity at 55 °C for more than 1 h at pH 6.4. Crude enzyme of the strain rich in levanase was established for the hydrolysis of levan in order to produce fructooligosaccharides with variable degrees of polymerization which could be used in important fields such medicine, food-processing industry and cosmetic. The extracellular levanase of the strain was then, partially purified as determined by SDS-PAGE. The purification was achieved by ammonium sulfate precipitation, gel filtration and DEAE cellulose chromatographies.

The extracellular fructosyltransferase (FTase) of a novel strain of Bacillus licheniformis capable of producing fructooligosaccharides (FOS) and a polysaccharide type levan was obtained and partially purified. The purification was achieved by ammonium sulfate precipitation, DEAE cellulose and gel filtration chromatographies. The enzyme was partially purified as determined by SDS-PAGE, and the specific activity reached was 67.5, representing a purification factor of 177 and yield of 40%. Levan was isolated from the cultures of B. licheniformis . The levan was composed mainly of fructose residues when analyzed by TLC after acid hydrolysis and NMR analysis. In a previous study, the levan produced exhibited a hypoglycemiant activity. The present paper deals with the study of the antitumor and anti-cytotoxic effect of levan produced by B. licheniformis strain. In the in vitro antitumor activity test of levan against some tumor cell lines, relatively the significantly high activity was observed against the HepG 2 .

Levan polysaccharide, a type of fructan, has been shown to have industrial applications as a new industrial gum in the fields of cosmetics, foods like dietary fiber and pharmaceutical goods. The objective of this current study was to investigate the possible hypolipidemic and antioxidative effects of levan in rats fed with a high-cholesterol diet. Animals were allocated into four groups of six rats each: a normal diet group (Control), normal rats received levan (L), a high-cholesterol diet group (Chol) and a high-cholesterol diet with a daily dose of levan equivalent to 5%. Treated hypercholesterolemic rats were administrated with levan in drinking water through oral gavage for 60 days. After the treatment period, the plasma antioxidant enzymes and lipid profiles were determined. Our results show that treatment with levan polysaccharide positively changed plasma antioxidant enzyme activities and lipid profiles (total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides) in cholesterol-rats, and thus may have potential hypolipidemic and antioxidant effects. Levan could protect against oxidative stress linked atherosclerosis and decrease the atherogenic index.

A newly isolated thermophilic bacterial strain from Tunisian thermal source was identified as Bacillus sp. and was selected for its ability to produce extracellular levansucrase. Following the optimization of carbon source, nitrogen source, temperature and initial pH of the growth medium in submerged liquid cultures. In fact, sucrose was found to be a good inducer of levansucrase enzymes. The optimal temperature and pH of the levansucrase were 50°C and 6.5, respectively and its activity increased four folds in the presence of 50mM Fe(2+). This enzyme exhibited a remarkable stability and retained 100% of its original activity at 50°C for more than 1h at pH 6.5. The half-life of the enzyme was 1h at 90°C. Crude enzyme of Bacillus sp. rich in levansucrase was established for the synthesis of fructooligosaccharides and levan. Bacillus sp. could therefore be considered as a satisfactory and promising producer of thermostable levansucrases. Contrary to other levansucrases, the one presented in the current study was able to produce high levels of levan with high molecular weight at 50°C and having an important effect as a hypoglycemic agent which was demonstrated in our previous publications (Dahech et al., 2011 [25]) and as a hypo-cholesterolemic agent which will be investigated in further research.

Diabetes is a serious condition that is linked to the development of oxidative stress. In the context of enhancing the biodiversity of Tunisia's flora, this study aimed to evaluate the effect of Sargussum vulgare sulfated polysaccharide (SVSP) on hyperglycemia and diabetes complications in the alloxan-induced diabetic rats. Our results showed a disturbance of carbohydrate, lipid, hematological and histopathological parameters, an increase in the α-amylase enzyme activity and damage to the pancreatic, hepatic and renal tissues in rats rendered diabetic by alloxan. In contrast, treatment with SVSP resulted in a correction of fasting and postprandial blood glucose and HbA1c through inhibition of pancreatic α-amylase. We also noticed an improvement in hemogram parameters and an attenuation of the pancreatic oxidative stress markers as well as histological protections. On the other hand, the administration of SVSP to diabetic rats caused, besides the correction of glycemic and lipid parameters, a good defense against hepatotoxicity and nephrotoxicity of diabetes as well as good antioxidant status and histological protections.

The entire pectate lyase cDNA (Pel1) of Penicillium occitanis was cloned from a cDNA bank and sequenced. The ORF exhibited a great homology to Penicillium marneffei and conservation of all features of fungal pectate lyases such as the barrel structure with "eight right-handed parallel β-helix" architecture. The structure modeling also showed the interesting resemblance with thermostable pectate lyases since several specific residues were also shared by Pel1 and these thermostable enzymes. Having shown that the enzyme retains its activity after endoH-mediated deglycosylation, we investigated its expression in Escherichia coli BL21 using the pET28-a vector. This expression was shown to be optimum when cells were induced at room temperature in 2YT medium rather than at 37 °C and LB medium. In such conditions, the recombinant protein was apparently produced more in soluble form than as inclusion bodies. The effect of NaCl concentration was investigated during the binding and elution steps of recombinant His-tagged enzyme on MagneHis Ni-particles. The purified enzyme was shown to retain its thermo-activity as well as a great tolerance to high concentration of NaCl and imidazole.

Crude extracts from the fruit of Lycium shawii, a plant collected from the south of Tunisia, were screened for their in vitro antioxidant, antimicrobial and antifungal activities. The dried fruits were extracted with EtOH and an aqueous suspension of the obtained EtOH extract was partitioned successively with CHCl3, CH2Cl2, EtOAc and n-BuOH, leaving residual water extract. Total phenolic content of extracts from these fruits were also determined. β-Carotene bleaching assay and Folin-Ciocalteu reagent were used to determine total antioxidant activity and total phenols of fruit extracts. Total phenolic and flavonoid contents varied from 100 to 377 mg GAE/g DW and 3.3-110.6 mg quercetin/gDW, respectively. Several extracts showed high antioxidant capacity and an antimicrobial activity against different strains. This is, to our knowledge, the first report on the detail chemical composition, antioxidant and antimicrobial activities of L. shawii extracts. The results provided evidence that the studied fruit might indeed be potential sources of natural antioxidant and antimicrobial agents.

Levan polysaccharide, a type of fructan, has been shown to favorably affect diabetes type 2 and hypercholesterolemia. Recent reports have indicated that excessive oxidative stress contributes to the development of atherosclerosis linked metabolic syndrome. The objective of this current study was to investigate the possible protection against oxidative stress linked atherosclerosis. A group of twenty four male rats was divided into four subgroups; a normal diet group (Control), normal rats received levan (L), a high-cholesterol diet group (Chol) and a high-cholesterol diet with 5% (w/w) levan group. After the treatment period, the plasma antioxidant enzymes and lipid profiles were determined. Our results show that treatment with levan positively changed plasma antioxidant enzyme activities by increasing superoxide dismutase (SOD) and catalase (CAT) by 40% and 28%, respectively, in heart. Similarly, the treatment of Chol fed groups with levan positively changed lipid profiles by decreasing total cholesterol, triglycerides and LDL-cholesterol by 50%, 38.33% and 64%, respectively. Thus may have potential antioxidant effects and could protect against oxidative stress linked atherosclerosis.

A strain of Bacillus licheniformis with high exopolysaccharide (EPS) production ability was isolated and identified. A new type of EPS was isolated from the strain fermentation and its structural characteristics were investigated and elucidated by partial and total acid hydrolysis, Fourier transform infrared, and (1)H and (13)C NMR spectroscopy including 2D (1)H, COSY, NOESY, XHCOR and HMBC experiments. Based on obtained data, the EPS was found to be a levan composed of linear chains of (2→6)-linked β-d-fructofuranosyl residues with connections β (2→6).

This study aimed to evaluate the effect of a polysaccharide named levan, which was produced by new isolated bacteria, on oxidative stress and hyperglycemia in alloxan-induced diabetic rats. Levan polysaccharide was given in drinking water for 60 days at a daily dose equivalent to 2%. The oral administration of levan in diabetic rats caused a decrease in glucose level in plasma and an increase of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities in both pancreas and liver. Furthermore, a protective action against hepatic and pancreatic toxicity in diabetic rats was clearly observed. Furthermore, a significant decrease in hepatic and pancreatic indices toxicity was observed, i.e., alkalines phosphatases (ALP), aspartate and lactate transaminases (AST and ALT), lactate deshydrogenases (LDH) activities and the thiobarbituric acid-reactive substances (TBARs). These beneficial effects of levan were confirmed by histological findings in hepatic and pancreatic tissues of diabetic rats. This study demonstrates for the first time that levan is efficient in inhibiting hyperglycemia and oxidative stress induced by diabetes and suggests that administration of levan may be helpful in the prevention of diabetic complications associated with oxidative stress.

This study aims to examine the effects of polysaccharide levan on oxidative stress and hyperglycemia in alloxan-induced diabetic rats. Levan, used in this study, was a microbial levan synthetisized by a non pathogenic bacteria recently isolated and identified as Bacillus licheniformis. Animals were allocated into four groups of six rats each: a control group (Control), diabetic group (Diab.), normal rats received levan (L) and diabetic rats fed with levan (DL). Treated diabetic rats were administrated with levan in drinking water through oral gavage for 60 days. The administration of polysaccharide levan in diabetic rats caused a significant increase in glycogen level by 52% and a decrease in glucose level in plasma by 52%. Similarly, the administration of polysaccharide levan in diabetic rats caused a decrease in the thiobarbituric acid-reactive substances (TBARS) by 31%, 41%, 39% and 25%, an increase in superoxide dismutase (SOD) by 40%, 50%, 44% and 34%, and in catalase (CAT) by 18%, 20%, 12% and 18% in liver, kidney, pancreas and heart, respectively. Furthermore, a significant decrease in hepatic and renal indices toxicity was observed, i.e. alkalines phosphatases (ALP), aspartate and lactate transaminases (AST and ALT) activities, total bilirubin, creatinine and urea levels by 19%, 31%, 32%, 36%, 37% and 23%, respectively. The results show that administration of polysaccharide levan can restore abnormal oxidative indice near normal levels. This study demonstrates, for the first time, that polysaccharide levan is efficient in inhibiting hyperglycemia and oxidative stress induced by diabetes and suggests that levan supplemented to diet may be helpful in preventing diabetic complications in adult rats.

The Talaromyces thermophilus strain produces only one form of lipase called TTLI. When the culture medium was concentrated and stored at 4 °C during a few days, we noticed the appearance of a second short form of lipase named TTLII. This second form was purified to homogeneity using gel filtration and FPLC-Anion exchange chromatography. The NH2-terminal 24 amino acid residues were found to be identical to those of TTLI. The treatment of the TTLI with endoglycosidase H decreased its apparent molecular weight from 39 to 30 kDa which corresponds to the molecular weight of TTLII. This difference was mostly attributed to the N-glycosylation of the enzyme. In fact, the glycan chain content and concavaline A–Sepharose affinity column confirmed that the TTLII was completely deglycosylated. Compared to TTLI, the TTLII activity was completely decreased over a broad range of temperature and pH. Furthermore, the deglycosylation of the enzyme reduced its specific activity by 50% toward different substrates; strongly suggest that the N-glycans are determinants for optimal catalytic activity and thermal stability of this enzyme. Covalent immobilization of the enzymes on supports suggests the involvement of the glycan moiety in enzyme–polymer interactions. In the case of TTLI the glycan moiety can constitute an extra site for the covalent linkage of the enzyme on the carrier.