Streptomyces flavogriseus HS1: isolation and characterization of extracellular proteases and their compatibility with laundry detergents.

The present study describes the isolation of a new protease producing Streptomyces strain HS1 and the biochemical characterization of the secreted proteases. By sequencing of its noted 16S rDNA, HS1 strain was found to have a 100% identity with Streptomyces flavogriseus. The highest protease production was found using FermII media. In these conditions maximum protease production (99 U/mL) was obtained after 96 h incubation at 30°C and 150 rpm. HS1 strain produced at least five proteases as revealed by zymogram technique. The enzyme preparation exhibited activity over a broad range of pH (5-11) and temperature (25-70°C). Optimum activity was observed at a pH of 7.0 and a temperature of 50°C. Proteolytic activity was significantly unaffected by Ca(2+) and Mg(2+). EDTA and PMSF highly decreased the original activity. The crude extracellular proteases showed high stability when used as a detergent additive. These properties offer an interesting potential for enzymatic hydrolysis at the industrial level.

Physical, structural, antioxidant and antimicrobial properties of gelatin-chitosan composite edible films.

Physico-chemical and mechanical properties of cuttlefish skin gelatin (G), chitosan (C) from shrimp (Penaeus kerathurus) and composite films (G75/C25, G50/C50, G25/C75) plasticized with glycerol were investigated. The results indicated that chitosan film had higher tensile strength and lower elongation at break when compared with the other films. Composite films show no significant difference in tensile strength (TS), thickness and transparency. The structural properties evaluated by FTIR and DSC showed total miscibility between both polymers. DSC scans showed that the increase of chitosan content in the composite films increases the transition temperature (Tg) and enthalpy (ΔHg) of films. The morphology study of gelatin, chitosan and composite films showed a compact and homogenous structure. In addition, gelatin and G75/C25 films demonstrated a high antioxidant activities monitored by ß-carotene bleaching, DPPH radical-scavenging and reducing power activities, while films contained chitosan exhibited higher antimicrobial activity against Gram-positive than Gram-negative bacteria.

Biological properties and biodegradation studies of chitosan biofilms plasticized with PEG and glycerol.

Chitosan biofilms, prepared by casting method at various percentage of plasticizer (PEG and glycerol), were evaluated for their biological, structural and thermal properties. The addition of PEG at 30% (w/w) and glycerol at 10% (w/w) to chitosan has increased the antioxidant activity of biofilm with the percentages of 22 and 26%, respectively. The increase of ferric reducing power was noted for the mixtures of chitosan-PEG (70-30) and chitosan-GLY (75-25). Additionally, the antibacterial properties of several biofilms were tested against E. coli and S. aureus. Biofilms with 70-30 and 90-10 blends ratio of chitosan-PEG and chitosan-GLY showed the best inhibitory effect against E. coli and S. aureus with 12 and 27%, respectively. All biofilms were degraded in compost in liquid and the addition of plasticizer PEG to chitosan increased his biodegradability with a value of BOD5 about 2.33 O2/mg CO. FT-IR spectra showed that the addition of plasticizer promoted the interactions through hydrogen bonding as reflected on the shifting of main peaks but there is no effect on biodegradation.

Comparative study of physico-mechanical and antioxidant properties of edible gelatin films from the skin of cuttlefish.

Physicochemical properties of edible films based on cuttlefish skin gelatin extracted without (G0) or with different concentrations of pepsins (5 (G5), 10 (G10) and 15 (G15) U/g of skin) were investigated. Edible films prepared with partially hydrolyzed gelatins had lower tensile strength (TS) and elongation at break (EAB), but higher water vapour permeability (WVP) and water solubility than the control film. FTIR spectra of obtained gelatin films revealed a significant loss of molecular order of the triple helix. In addition, differential scanning calorimetric (DSC) analysis indicated that partially hydrolyzed gelatine films exhibited lower transition temperature and enthalpy compared with those of control film. The properties of the films were related to their microstructure, which was observed by scanning electron microscopy. Films with G0 and G5 had a smooth surface and a more compact structure, while films prepared with G10 and G15 had coarser surface. Thus, the chain length of extracted gelatin directly affected the properties of corresponding films.

In vitro study of the PLA2 inhibition and antioxidant activities of Aloe vera leaf skin extracts.

BACKGROUND: In the present work we determined the total phenolic content of Aloe vera leaf skin (AVLS) extracts by using various solvents (hexane, chloroform-ethanol (1/1), ethyl acetate, butanol and water). We have also evaluated the antioxidant and the anti-PLA2 properties of these extracts by measuring their inhibition potency on the human pro-inflammatory phospholipase A2 (group IIA). RESULTS: The water extract exhibits the highest inhibitory effect with an IC50 = 0.22 mg/ml and interestingly no effect was observed on the digestive phospholipase A2 (group IB) even at a concentration of 5 mg/ml. Antioxidant activities were also analyzed and the most active extracts were observed when using chloroform ethanol (1/1) and ethyl acetate (IC50 = 0.274 and 0.326 mg/ml, respectively). Analysis of the total phenolic content reveals that the water extract, with the best anti-PLA2 effect, was poor in phenolic molecules (2 mg GAE/g). This latter value has to be compared with the chloroform-ethanol and the ethyl acetate extracts (40 and 23.8 mg GAE/g, respectively), mostly responsible for the antioxidant activity. CONCLUSION: A significant correlation was established between the total phenolic content and the antioxidant capacity but not with the anti PLA2 activity. Results from phytochemical screening suggest that the anti PLA2 molecules were probably catechin tannins compounds.

Inhibition of pro-inflammatory secreted phospholipase A2 by extracts from Cynara cardunculus L.

The aim of the present work was to evaluate the anti-inflammatory properties of Cynara cardunculus L. (Asteraceae) during its growth using various solvents such as n-hexane, dichloromethane, acetone, and methanol for air-dried leaves and stems. The anti-inflammatory activities of crude extracts were evaluated by measuring the inhibition potency of mammalian non-pancreatic phospholipases A2 (hG-IIA). The methanol and acetone extracts of leaves harvested in February exhibit potent inhibition of hG-IIA (IC(50) = 50 and 70 microg/ml, respectively). However, the acetone extract of stems harvested in December inhibits the hG-IIA with a lower IC(50) around 130 microg/ml. Fractionation on silica gel and hydrophobic gel of the methanol extract of leaves harvested in February increases the inhibitory effect, and the IC(50) reached 10 microg/ml.

Purification and characterization of the first reptile pancreatic lipase: the turtle.

OBJECTIVES: The aim of the present study was to purify and characterize classic pancreatic lipase from the reptile turtle (TuPL). METHODS: The lipase was purified from the fresh pancreas extract followed by diethylamino ethyl-cellulose chromatography, Sephacryl S-200 gel filtration, and a Mono-Q Sepharose chromatography. RESULTS: Turtle pancreatic lipase is a serine enzyme and it contains only 1 free cysteine. Its activity is maximum at pH 8.2 and 37 degrees C. A specific activity of 10.000 U/mg and 5.000 U/mg were measured titrimetrically on tributyrin and olive oil emulsion, respectively. Natural detergents act as potent inhibitors of TuPL, and colipase restores the activity. When the lipase is inhibited by synthetic detergent, simultaneous addition of colipase and bile salts is required to restore the TuPL activity. The critical surface pressure of TuPL (pi(c)) = 20.9 mN m(-1)) is similar to the one of human PL (pi(c) = 18 mN m(-1)). CONCLUSIONS: The results presented in this article indicate that despite the primitive character of the turtle, no significant difference has been observed between TuPL and known mammalian PLs. However, partial proteolysis of TuPL with chymotrypsin shows the absence of the 14-kDa fragment identified as the C-terminal domain in the case of many classic PLs.

Purification and biochemical characterization of a turkey preduodenal esterase.

A preduodenal esterase was purified to homogeneity from turkey (Meleagris gallopavo) pharyngial tissue. Pure turkey pregastric esterase (TPrE) was obtained after anion exchange chromatography (DEAE-cellulose), Sephacryl S-200 gel filtration, anion exchange chromatography (Mono-Q sepharose) and affinity chromatography (Blue-Gel Affi Gel). The pure enzyme has an apparent molecular mass of 50 kDa, as determined by SDS/PAGE analysis. The optimum pH and temperature for enzyme activity using tributyrin as substrate were 8.5 and 48 degrees C, respectively. Under these conditions, the specific activity measured was 650 U/mg. No significant lipolytic activity was found when was tested on triolein or liprocil as substrates or with monolayer dicaprin with TPrE. In contrast, TPrE displayed a maximal activities of 800, 680 and 520 U/mg with vinyl acetate, vinyl propionate and vinyl butyrate, respectively. This enzyme retained 75% of its maximal activity when incubated for 30 min at pH 4 and 50 degrees C, but it was completely inactivated after an incubation for 10 min at 60 degrees C. The TPrE N-terminal amino acid sequence showed similarities to the N-terminal sequence of a thioesterase from mallard duck, but no similarity with known preduodenal lipases was found.