Selection of Lactobacillus plantarum TN627 as a New Probiotic Candidate Based on in vitro Functional Properties

Wacim Bejar,Ameny Farhat-Khemakhem,Slim Smaoui,Mohamed Makni,Mounira Ben Farhat,Badis Abdelmalek,Lotfi Mellouli,Samir Bejar,Hichem Chouayekh,Emmanuelle Maguin 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.6

Nine lactobacilli previously selected for high antagonism against food borne bacterial pathogens were identified via 16S rRNA gene sequencing and screened for probiotic potential for use in poultry production. The lactobacilli were subjected to a subtractive in vitro analysis system using a certified probiotic as reference. This allowed for selection of a milk-derived Lactobacillus plantarum strain, termed TN627. This organic acid-producing bacterium was free of harmful enzymatic activity and sensitive to several antibiotics. It also showed good growth at pH 4 and in the presence of bile. L. plantarum TN627 also exhibited high efficacy of adhesion to chicken enterocytes, which correlated with detecting genes encoding the mucusbinding,adhesion-promoting proteins (Mub and MapA)and the adhesion-like factor EF-Tu, commonly involved in adherence of lactobacilli to mucosal surfaces. Taken together, our findings suggest that TN627 is a promising probiotic candidate with high potential for application as a supplement in the animal feed industry.

Cucurbituril complexes cross the cell membrane

Montes-Navajas, Pedro,Gonzalez-Bejar, Maria,Scaiano, J.C.,Garcia, Hermenegildo Korean Society of Photoscience 2009 Photochemical & photobiological sciences Vol.8 No.12

Cucurbiturils (CBs) of the appropriate size (CB[7] and CB[8]) form strong guest-host complexes in phosphate buffer solution (PBS) with acridine orange (AO) and pyronine Y (PYY) with 1 : 1 and 2 : 1 stoichiometries for CB[7] and CB[8] complexes, respectively. Binding constants in the range $0.87-1.60{\times}10^6M^{-1}$ and $5.2-6.3{\times}10^{13}M^{-2}$ were determined by titration with fluorescence spectroscopy for 1 : 1 and 2 : 1 complexes, respectively. These binding constants in PBS and the eight-fold excess of CBs minimize the presence of free dye in solution and also stabilize the host-guest complex in the culture medium. Images showing that the CB complexes can cross the cell membrane of 3T3 cells have been acquired using fluorescence microscopy. Given the current importance of supramolecular CB complexes and the search for new drug delivery systems, the present findings open avenues for the use of CBs as nanocapsules to transport drugs into the cells.

Optimization of Submerged Aspergillus oryzae S2 α-Amylase Production

Belgacem Naili,Mouna Sahnoun,Samir Bejar,Radhouane Kammoun 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.1

Use of 4 agro-industrial by products and organic materials as nitrogen sources for production of Aspergillus oryzae S2 α-amylase in liquid culture was investigated. The 2 agro-industrial byproducts maltose and saccharose, and also lactose and starch were individually evaluated for use as carbon sources. A Box-Behnken experimental design was used to determine optimal conditions for production of α-amylase. A maximum amylase activity of 750 U/mL was obtained at a temperature of 24oC, a urea concentration of 1 g/L, and a C/N ratio of 2. Laboratory scale application of optimal conditions in a 7 L fermentor produced a final α-amylase activity of 770 U/mL after 3 days of batch cultivation. Addition of 10% starch to the culture medium each 12 h immediately after the stationary phase of cell growth led to a production yield of 1,220 U/mL at the end of fed-batch cultivation.

Modulatory Effect of Fenugreek Saponins on the Activities of Intestinal and Hepatic Disaccharidase and Glycogen and Liver Function of Diabetic Rats

Khaled Hamden,Abdelfattah Elfeki,Bassem Jaouadi,Samir Bejar,Tahia Salami,Serge Carreau 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.5

Diabetes mellitus is a serious health concern throughout the world and is often associated with a variety of bodily disorders such as liver toxicity and dysfunction. This study elucidates the effect of fenugreek saponin administration on disaccharidase and glycogen activities in the intestine and liver of surviving diabetic rats. It also evaluates the effect of saponin feeding using a number of liver toxicity indices, namely stress oxidant, liver dysfunction markers and metabolism. Our findings indicate that the fenugreek saponin fraction significantly modulated the disaccharidase and glycogen enzyme activities in the intestine and liver of rats, increased the hepatic glycogen content, suppressed the increase of blood glucose level and improved results in the oral glucose tolerance test (OGTT). The fenugreek saponin extract also efficiently protected the hepatic function, which was evidenced by the significant increases of superoxide dismutase (SOD), catalase (CAT),gluthation peroxidase (GPX), aspartate transaminase (AST),alanine transaminase (ALT) and lactate deshydrogenase (LDH) enzyme activities. Fenugreek saponin also induced a notable delay in the absorption of LDL-cholesterol and triglycerides and a remarkable increase in levels of HDLcholesterol. A histological analysis of the hepatic tissues further established the positive effect of fenugreek saponin. Overall, the findings of the current study indicate that fenugreek saponins exhibit attractive properties and can be considered as promising candidates for future application as therapeutic agents in biotechnological and bioprocessbased technologies, particularly those related to the development of anti-diabetic, hepatoprotective and hypolipidemic drugs.

Magnetic and Magnetocaloric Properties of La<SUB>0.7</SUB>(CaSr)<SUB>0.3</SUB>Mn<SUB>1-x</SUB>Ga<SUB>x</SUB>MnO₃ (x=0.00, 0.025, 0.05, 0.075 and 0.10) Compounds

S. Othmani,A. Tozri,M. Bejar,E. Dhahri,E. K. Hlil 한국자기학회 2008 한국자기학회 학술연구발표회 논문개요집 Vol.- No.-

Inhibitory Effect of Fenugreek Galactomannan on Digestive Enzymes Related to Diabetes, Hyperlipidemia, and Liver-kidney Dysfunctions

Khaled Hamden,Abdelfattah Elfeki,Bassem Jaouadi,Samir Bejar,Serge Carreau 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.3

The present study was undertaken to investigate the effect of fenugreek galactomannan on intestinal glucose uptake in surviving diabetic rats. It explored their potential action with respect to lowering maltase, lactase,and sucrase activities in the small intestine of galactomannan-treated diabetic group compared to the diabetic control group. The findings indicate that the increase of blood glucose levels was significantly suppressed in the galactomannan-treated group than those in the diabetic rats. Moreover, the galactomannan isolated from fenugreek exhibited a prominent selective inhibitory effect against intestinal lipase activity. It was found to significantly delay the absorption of LDL-cholesterol and triglycerides and the increase in HDL-cholesterol. In addition, fenugreek galactomannan efficiently protect the hepatic function observed by the considerable decrease of aspartate and alanine transaminases (AST and ALT) and lactate deshydrogenase (LDH) contents in the serum of diabetic rats. The beneficial effects of fenugreek galactomannan were also evidenc-ed by their capacity to inhibit diabetes-induced kidney injury through lowering the urea and creatinine content in plasma. Overall, the conclusion of the present study indicate that fenugreek galactomannan displays a number of promising properties and attributes for future applications as therapeutic agents in biotechnological and bioprocess-based technologies, particularly those interested in the development of anti-diabetic and hypolipidemic drugs.

Optimization of Aspergillus oryzae S2 α-amylase, ascorbic acid, and glucose oxidase combination for improved French and composite Ukrainian wheat dough properties and bread quality using a mixture design approach

Sahnoun, Mouna,Kriaa, Mouna,Besbes, Souhail,Jardak, Mohamed,Bejar, Samir,Kammoun, Radhouane 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.5

A simplex-centroid experimental design was used for the optimization of both reducing and oxidizing improvers, namely Aspergillus oryzae S2 ${\alpha}$-amylase (Amy), ascorbic acid (Asc), and glucose oxidase (GOD). This optimization was performed to enhance the dough and breadmaking qualities of soft French wheat flour and a composite counterpart that contained 30% Ukrainian wheat flour. Statistically significant correlations were calculated between the W index and textural parameters (e.g., dough chewiness and bread cohesiveness). The findings revealed that while the best mixture for French flour comprised 21.8% of Amy, 41.2% of Asc, and 37% of GOD, for the composite counterpart, it comprised 2.3% of Amy, 66% of Asc, and 31.7% of GOD. These optimized mixtures rearranged soft French wheat flour and its composite counterpart to a good quality and an improved flour texture, respectively. Additionally, they increased the loaf specific volumes of the breads made from soft French wheat flour and its counterpart by 25.8 and 45.43%, respectively, significantly decreased the breads' susceptibility to microbial contamination, and reclassified the breads as "good" in terms of sensory attributes.

Bacillus subtilis Bacteriocin Bac 14B with a Broad Inhibitory Spectrum: Purification, Amino Acid Sequence Analysis, and Physicochemical Characterization

Ines Hammami,Bassem Jaouadi,Abir Ben Bacha,Ahmed Rebai,Samir Bejar,Xavier Nesme,Ali Rhouma 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.1

Bacillus subtilis strain 14B was used to produce a novel antimicrobial peptide (bacteriocin) called Bac 14B. Pure bacteriocin was obtained after heat and acidic treatments (80°C and pH 4), precipitation by ammonium sulfate, and chromatography on Sephadex G-50 and Mono Q Sepharose columns. Based on MALDI-TOF mass spectrometry analysis, purified Bac 14B is a monomer protein with a molecular mass of 20110.13 Da. N-terminal sequencing allowed for the straightforward identification of its first 12 residues, which were of a pure bacteriocin. It also revealed that this bacteriocin contained a unique sequence, namely M-L-K-A-N-L-Q-N-P-L-N-A, suggesting the identification of a novel compound. Bac 14B was stable for 1 h at temperatures up to 80°C and pH of 4 ~ 8. It also proved sensitive to various proteases, which demonstrated its protein nature. Bac 14B displayed a bacteriolytical mode of action and a broad range of inhibitory spectra toward Gram-positive and -negative pathogens. Interestingly,based on conventional agronomic seed vigor parameters,the application of Bac 14B (500 activity units/mL)to various crops revealed that this bacteriocin was a potent exogenous enhancer of growth that stimulated the seedling vigor of tomatoes and muskmelons. Compared to those of the control, the germination percentage, shoot weight,shoot height, and root length were all significantly enhanced in Bac 14B-treated plant seeds. Bac 14B also exhibited effective disinfectant properties against a wide range of seedborne diseases and significant effects on the control of damping off diseases, particularly at the pregermination stage. It also proved to be effective against root rot diseases caused by Alternaria solani and other bacterial seedborne pathogens such as wilt diseases. The findings indicate that Bac 14B is the first B. subtilisproduced bacteriocin ever reported to exhibit such promising biological properties. Bacillus subtilis strain 14B was used to produce a novel antimicrobial peptide (bacteriocin) called Bac 14B. Pure bacteriocin was obtained after heat and acidic treatments (80°C and pH 4), precipitation by ammonium sulfate, and chromatography on Sephadex G-50 and Mono Q Sepharose columns. Based on MALDI-TOF mass spectrometry analysis, purified Bac 14B is a monomer protein with a molecular mass of 20110.13 Da. N-terminal sequencing allowed for the straightforward identification of its first 12 residues, which were of a pure bacteriocin. It also revealed that this bacteriocin contained a unique sequence, namely M-L-K-A-N-L-Q-N-P-L-N-A, suggesting the identification of a novel compound. Bac 14B was stable for 1 h at temperatures up to 80°C and pH of 4 ~ 8. It also proved sensitive to various proteases, which demonstrated its protein nature. Bac 14B displayed a bacteriolytical mode of action and a broad range of inhibitory spectra toward Gram-positive and -negative pathogens. Interestingly,based on conventional agronomic seed vigor parameters,the application of Bac 14B (500 activity units/mL)to various crops revealed that this bacteriocin was a potent exogenous enhancer of growth that stimulated the seedling vigor of tomatoes and muskmelons. Compared to those of the control, the germination percentage, shoot weight,shoot height, and root length were all significantly enhanced in Bac 14B-treated plant seeds. Bac 14B also exhibited effective disinfectant properties against a wide range of seedborne diseases and significant effects on the control of damping off diseases, particularly at the pregermination stage. It also proved to be effective against root rot diseases caused by Alternaria solani and other bacterial seedborne pathogens such as wilt diseases. The findings indicate that Bac 14B is the first B. subtilisproduced bacteriocin ever reported to exhibit such promising biological properties.

Optimization of Aspergillus oryzae S2 α-amylase, ascorbic acid, and glucose oxidase combination for improved French and composite Ukrainian wheat dough properties and bread quality using a mixture design approach

Mouna Sahnoun,Mouna Kriaa,Souhail Besbes,Mohamed Jardak,Samir Bejar,Radhouane Kammoun 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.5

A simplex-centroid experimental design was used for the optimization of both reducing and oxidizing improvers, namely Aspergillus oryzae S2 α-amylase (Amy), ascorbic acid (Asc), and glucose oxidase (GOD). This optimization was performed to enhance the dough and breadmaking qualities of soft French wheat flour and a composite counterpart that contained 30% Ukrainian wheat flour. Statistically significant correlations were calculated between the W index and textural parameters (e.g., dough chewiness and bread cohesiveness). The findings revealed that while the best mixture for French flour comprised 21.8% of Amy, 41.2% of Asc, and 37% of GOD, for the composite counterpart, it comprised 2.3% of Amy, 66% of Asc, and 31.7% of GOD. These optimized mixtures rearranged soft French wheat flour and its composite counterpart to a good quality and an improved flour texture, respectively. Additionally, they increased the loaf specific volumes of the breads made from soft French wheat flour and its counterpart by 25.8 and 45.43%, respectively, significantly decreased the breads’ susceptibility to microbial contamination, and reclassified the breads as “good” in terms of sensory attributes.

Excellent Laundry Detergent Compatibility and High Dehairing Ability of the Bacillus pumilus CBS Alkaline Proteinase (SAPB)

Bassem Jaouadi,Semia Ellouz-Chaabouni,Mamdouh Ben Ali,Ezzedine Ben Messaoud,Belgacem Naili,Abdelhafidh Dhouib,Samir Bejar 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.4

The newly Tunisian soil-isolated bacterium, producing the alkaline proteinase termed SAPB that was already purified and characterized [1], was assigned as Bacillus pumilus CBS strain on the basis of biochemical properties and 16S rRNA gene sequencing. The maximum protease activity recorded after 24 h of incubation in an optimized medium at 37°C was 6,500 U/mL in shaking flask culture and 25,000 U/mL in fermentor. SAPB showed excellent stability and compatibility in laundry detergent retaining more than 98% of its initial activity after pre-incubation for 1 h at 40°C with Det, followed by OMO (97%), Dinol (94%), and Dixan (93%). Examination of various stained cloth pieces exhibited a remarkable efficiency in the removal of blood and chocolate stains. More interestingly, SAPB demonstrated powerful dehairing capabilities of hair removal from skin with minimal damage on the collagen and a nearly complete feather-degradation. Likewise, Bacillus pumilus CBS effectively degraded feather-meal (98.5%), chicken feather (92%), goat hair (80%), and bovine hair (68%) whereas sheep wool under went less degradation. Keratin-degradation resulted in sulfhdryl group formation (0.95~3.91 μM).