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Open Access Article
1 - Molecular identification of subtilisin genes (SUB3 and SUB6) in Epidermophyton floccosum
, E. Khedmati , SJ. Hashemi Hazaveh , M. Bayat , K. AminiEpidermophyton floccosum is one of the worldwide anthropophilic dermatophytes that invade keratinized structures such as hair, skin, and nail, causing dermatophytosis by secreting important proteases such as subtilisin. This study aimed to evaluate the presence of SUB3 MoreEpidermophyton floccosum is one of the worldwide anthropophilic dermatophytes that invade keratinized structures such as hair, skin, and nail, causing dermatophytosis by secreting important proteases such as subtilisin. This study aimed to evaluate the presence of SUB3 and SUB6 encoding serine proteases in the isolate, which received from the fungi culture collection of Tehran University of Medical Sciences, Faculty of Public Health. Special primers designed according to the highly conserved regions of similar genes in other dermatophytes. Genomic DNA and designed primers used in PCR, then PCR products sequenced with ABI PRISM®3730XL automated Sanger sequencer, and presence of 2 new subtilisin genes (SUB3 and SUB6) were confirmed and recorded in NCBI(with the accession numbers MN206114, MN177931 respectively). The coding sequence of SUB3 found to contain 861 nucleotides, which encodes a polypeptide with 287 amino acids. The coding sequence of SUB6 found to contain 699 nucleotides that encode a polypeptide with 233amino acids. Comparing the sequences with Gene Bank database information, revealed significant homology with other dermatophytes. Achieving a better understanding of the molecular characteristics of virulence genes may help develop effective therapies and prevention strategies. Manuscript profile -
Open Access Article
2 - Optimisation of the Conditions for Using Protease and Hemicellulase in Wafer Formulation
Babak Ghiassi Tarzi Venushe Babaei Nami Parviz Shahbazikhah Haleh HadaeghIntroduction: Economic issues concerned with the demand for energy reduction and costs as well as improved quality of the final product in the bakery industry such as Wafer production is of great importance.Materials and Methods: In the present study the effects of appl MoreIntroduction: Economic issues concerned with the demand for energy reduction and costs as well as improved quality of the final product in the bakery industry such as Wafer production is of great importance.Materials and Methods: In the present study the effects of applying different amounts of Protease (0.005-0.05% of the weight of flour), Hemicellulase (0.005-0.05% of the weight of flour), water reduction (0-20%), dough resting time (0-30 min) and the baking time (90-150 sec) on decreasing the waferbatter viscosity and some quality factors of wafer sheet were investigated and compared with the control sample. The amounts of batter viscosity, moisture, crispness and colour of wafer sheet were considered in order to evaluate the effects of various levels of protease and hemicellulas enzymes on the mentioned parameters. Optimisation of the process condition was conducted to determine the optimum formulation for producingwafer sheet.Results: The results were analyzed by Response Surface Methodology (RSM) and showed that optimal level of Protease enzyme, Hemicellulase enzyme, water reduction of formulation, dough resting time and baking time were 0.05%, 0.05%, 9.58%, 30 min and 135.17 sec, respectively.Conclusion: Protease and hemicellulase enzymes might be used in wafer sheet formulation and these enzymes have positive effects on dough and some quality factors of the final product such as moisture and crispiness. According to the variation of the flour quality, the production of wafer will beless dependenton the flour quality. Manuscript profile -
Open Access Article
3 - Contribution of the lycotetraose moiety of α-tomatine to the interaction with the main proteases of coronaviruses PEDV and SARS-CoV-2
Gerard Vergoten Christian Bailly -
Open Access Article
4 - Milk Enzymes
Mandana Zormand mahnaz hashemi ravanMore than 60 types of endogenous enzymes of milk have been identified in the milk of different species of mammals so far. This study has investigated milk enzymes. Milk clotting enzymes are made from different sources that rennet with animal origin is the most important MoreMore than 60 types of endogenous enzymes of milk have been identified in the milk of different species of mammals so far. This study has investigated milk enzymes. Milk clotting enzymes are made from different sources that rennet with animal origin is the most important and the oldest of all. But due to the reduction of its production, attempt has been made to use microbial, herbal and poultry substituents that such substituents must have acidic protease. There are some proteases in raw milk with low enzymatic activity which don’t play a role in coagulation because they cause B casein hydrolysate to Gama casein. But there are some psychrotroph gram-negative bacteria and some species of pseudomonas, alcaligenes and flavobacterim in raw milk that cause casein hydrolysate to para kappa-casein compounds and coagulation with their proteases. Milk enzymatic coagulation basically depends on the sensitivity of kappa-casein and the presence of calcium. After kappa-casein breaking at the presence of enzyme and disorganization of its structure, the micelles have accumulated and entangled into each other and form curd and coagulation occurs. Manuscript profile -
Open Access Article
5 - The effect of gibberellin hormone on yield, growth indices, and biochemical traits of corn (Zea Mays L.) under drought stress
Abbas Maleki Amin Fathi Sadegh BahaminIn order to investigate the effect of gibberellin hormone and drought stress on corn, an experiment was conducted as split plots based on a randomized complete block design with three replications. Treatments included moisture stress at three levels of normal irrigation MoreIn order to investigate the effect of gibberellin hormone and drought stress on corn, an experiment was conducted as split plots based on a randomized complete block design with three replications. Treatments included moisture stress at three levels of normal irrigation (I0=60), medium stress (I1=90), and severe stress (I2=120) mm evaporation from class A evaporation pan as main factor and a sub factor of gibberellin hormone spraying at four levels non-consumption (G0=0), (G1=15ppm), (G2=20ppm), and (G3=25ppm). Results showed that the highest and lowest grain yields were observed in 60 mm evaporation from evaporation pan with 20 ppm gibberellin acid and 120 mm evaporation from the evaporation pan without using gibberellin acid as 9658.2 and 5797.3 kg/h, respectively. Also, under all levels of drought stress, gibberellin acid application increased grain yield of corn. The interaction of hormones and irrigation had a significant effect on starch, proline, alpha amylase, beta amylase, and protease. Drought stress increased proline concentration in corn leaf. Gibberellin hormone had no significant effect on proline concentration in corn leaf under drought stress conditions, but under prolonged drought stress, proline concentration increased. The application of 20 and 25 ppm gibberellin under moderate dehydration (90 mm evaporation from evaporation pan) increased leaf proline concentrations by 36% and 50%, respectively, compared with control treatment (no gibberellin consumption). Under severe drought stress conditions (120 mm evaporation from the evaporation pan), proline concentrations with 20 and 25 ppm gibberellin were 32% and 21% higher than gibberellin consumption, respectively. Overall, the results showed that gibberellin acid, through positive effects on increasing and improving the yield components, can ultimately increase corn grain yield. Manuscript profile -
Open Access Article
6 - Detailed study and molecular identification of bacteria spp. causing bitter flavour in pasteurized cream
Hoda Dezhkhi Sajjad Yazdansetad Nazila Arbabsoleymani Reza Najafpour Seyed Mohammad Gheibi hayat الیکا فرج تبریزیCream enabled the growth of most microorganisms due to the adequate moisture, pH close to neutral, and nutrient rich. The major cause of bitter flavour in cream is degradation of proteins to the hydrophobic peptides by protease enzyme derived from the thermostable bacte MoreCream enabled the growth of most microorganisms due to the adequate moisture, pH close to neutral, and nutrient rich. The major cause of bitter flavour in cream is degradation of proteins to the hydrophobic peptides by protease enzyme derived from the thermostable bacteria resistant to the pasteurization temperature. The present study was aimed to investigate and molecular identify of bacteria spp. causing bitter flavour in pasteurized cream. The pasteurized cream samples were collected, then microbiological tests such as total count of microorganisms, psychrotrophic bacteria count, coliform bacteria count, thermostable bacteria count, sporogen bacteria count and also chemical tests such as acidity and pH measurement were carried out. The tests were fulfilled with three repetitions at 4ºC and 15ºC in first, third, fifth, seventh, ninth, and eleventh days after the cream production. The isolates causing bitter flavour in creams were identified by physiological, biochemical, phylogenetical, and molecular methods. The bacterial isolates were inoculated into the pasteurized creams and assayed for changing in flavour. The two species of bacteria causing bitter flavor were isolated from the pasteurized creams. The sequencing of 16S rRNA gene of isolates indicated that they were Bacillus cereus and Bacillus subtilis. The temperature of preservation is one of the most important agents of microorganisms’ activity and dairy product durability. Keeping raw milk for a long time before the pasteurization leads to increasing proteolytic bacteria resistant to the pasteurization temperature, producing protease by the organisms and finally, changing the product flavour. Manuscript profile -
Open Access Article
7 - The Effect of Protease Enzyme and Replacement Soybean Meal by Cotton Seed Meal on Protein Efficiency and Performance in Broiler Chickens
M. Safari A.A. Saki S. Mirzaie Goudarzi A. Ahmadi A. Ashoori -
Open Access Article
8 - Improvement of protease production by Chryseobacterium indologenes BYK27 and its application in de-colorization of blood on clothes
Yasamin Binabadi Arastoo Badoei-dalfard Abdolhamid Namaki-ShoushtariProtease is one of the most important industrial enzymes occupying nearly 60% of global enzyme sales. Extracellular protease finds numerous applications in industrial processes like in leather tanning, detergents, dairy, brewery as well as meat tenderization industries. MoreProtease is one of the most important industrial enzymes occupying nearly 60% of global enzyme sales. Extracellular protease finds numerous applications in industrial processes like in leather tanning, detergents, dairy, brewery as well as meat tenderization industries. In spite of that, the low level of enzyme production is the main challenge of industrial production of enzyme. Therefore, optimization of industrial protease production and its application in blood de-staining were the aims of this study. The sewage samples were cultivated on the skim milk agar. BYK27 isolates with the highest clear halo around the colonies were selected for further studies. Optimization of parameters affecting protease production by Chryseobacterium indologenes BYK27 was studied by Taguchi approach. De-staining ability of protease was also investigated by de-colorization of bloody cotton cloth. The optimal factors for protease production by Ch. indologenes BYK27 were found to be the temperature of 40 ˚C, pH of 9.0, 0.06% yeast extract and 1% glucose supplements. Protease production under optimal condition was found to be 590 (U/ml) which was improved by 63%, as compared to the basal medium. The protease activity and stability were increased 50% by beta-mercaptoethanol but inhibited about 88% by DMF. In addition, BYK27 protease was able to completely de-stain blood after 20 min of incubation. The results of this study indicate that BYK27 protease has biotechnological potential, specifically in the detergent industry and provision of valuable compounds. Manuscript profile -
Open Access Article
9 - Isolation and identification of moderately halophilic protease-producing bacteria from saline soils
Niloofar Ghorbani raz Maryam GhaneBackground & Objectives: Moderate halophiles are excellent sources of enzymes that are not only salt- stable, but also have optimal activities at a wide range of pH and temperature. The aim of this study was the isolation of moderately halophilic protease-producing MoreBackground & Objectives: Moderate halophiles are excellent sources of enzymes that are not only salt- stable, but also have optimal activities at a wide range of pH and temperature. The aim of this study was the isolation of moderately halophilic protease-producing bacteria from saline soils in Alborz province. Material & Methods: Screening of the protease- producing halophilic bacteria was carried out by skim milk agar containing 5% NaCl. Enzyme assay was investigated by the colorimetric method and the effect of various parameters such as temperature, pH, and different NaCl concentrations on enzyme activity were assessed. Identification of the isolates was carried out by biochemical as well as molecular methods. Results: A total of 11 halophilic strains with proteolytic activity were isolated among which the strain B8 with higher clearance zone on skim milk agar and 3.2 Unit/mL supernatant activity was chosen for further analysis. The enzyme exhibited its optimal activity at the temperature of 40◦C, pH of 7.5 and NaCl concentration of 0-0.5 M, although at higher salinities (up to 3 M) activity was still remained. The enzyme was active at a broad pH range, keeping 60% of its activity at pH of 9.5. Phylogenetic analysis based on 16S rRNA gene sequencing identified the isolate asHalomonas. It was deposited in GenBank, with the name of Halomonas sp. strain HM_NG2. Conclusion: These findings suggest that the protease secreted by the isolate of this study could be a good candidate for biotechnological applications due to its moderate thermo-stability and halloalculophytic properties. Manuscript profile -
Open Access Article
10 - Investigation of the effect of Bacillus licheniformis alkaline protease on qualitative, rheological and sensory properties of macaroni enriched with soy
Seyed Emad Hosseini Fatemeh ArdestaniBackground & Objectives: Macaroni is produced since 1313 in Iran, but with not desirable quality due to low accessibility of durum wheat and semolina production technology. In this study, we analysed the effect of Bacillus licheniformis alkaline protease on qualitat MoreBackground & Objectives: Macaroni is produced since 1313 in Iran, but with not desirable quality due to low accessibility of durum wheat and semolina production technology. In this study, we analysed the effect of Bacillus licheniformis alkaline protease on qualitative, rheological and sensory properties of macaroni enriched with soy. Materials & Methods: 1 kg of dough and dried macaroni was sampled based on the standard methods and used for evaluation of qualitative, rheological and sensory properties of macaroni enriched with hydrolyzed soy protein. Bacillus licheniformis was used because of its ability to produce an efficient alkaline protease for hydrolysis of soy proteins. Alkaline protease was produced in a submerged culture of Bacillus licheniformis at pH 9 and 300C for 3 days. Results: Addition of 20% hydrolyzed soy protein caused decrease in some factors including 15.78% in cooking time, 18.13% in resistance to defeat for a spaghetti filament, 29.02% in dough development time and 6.27% in loosening the dough as well as an increase of 37.37% in cooking loss, 57.24% in adhesion, 5.55% in water absorption percentage and 16.44% in dough stability time. Total acceptance of macaroni samples with 20% hydrolyzed soy protein were 33% less than control. The maximum useable hydrolyzed soy protein in macaroni dough without any negative impacts on its properties was determined as 5%. Conclusion: Hydrolyzed soy protein is not a suitable case for macaroni enrichment. Manuscript profile -
Open Access Article
11 - The production of alkaline protease by Bacillus tequilensis FJSH2 isolated from Jiroft’s slaughterhouse wastes
Arasto Badoei-dalfard Parvin Amiri Narjes Ramezanipour Zahra Karami Batool GhanbariBackground & Objectives: Proteases are the most applied industrial enzymes usable in several biotechnological industries. These enzymes include approximately 60% of enzyme marketing worldwide. Alkaline proteases are the most important enzymes which are stable in the MoreBackground & Objectives: Proteases are the most applied industrial enzymes usable in several biotechnological industries. These enzymes include approximately 60% of enzyme marketing worldwide. Alkaline proteases are the most important enzymes which are stable in the presence of alkaline pH, surfactant and oxidizing agents. This study was aimed to isolate and identify the protease producing bacteria from slaughterhouse wastes in Jiroft. Materials & Methods: This cross - sectional study was performed on Jiroft’s slaughterhouse wastes. The protease producing strain was screened on the skim milk agar media. The 16S rRNA genes of the samples were obtained using PCR and sequencing. The protease enzyme was partially purified by ammonium sulphate precipitation, dialysis and ion exchange chromatography. Thereafter, The biochemical features of the enzymes were analysed. Results: The gene sequencing study showed that the isolated strain were 100 % similar to Bacillus tequilensis species. The purified protease showed maximum activity and stability in pH 9 and 40°C. Km and Vmax values were obtained based on Linewewr berg equation 6.77 mg/ml and 64.94 U/ml/min, respectively. Protease stability was improved about 1.8 and 2.4 folds in the presence of cyclohexane and DMSO, respectively. Conclusion: The activity and stability of this enzyme in alkaline pH and in the presence of organic solvents indicates its potential to be used in industries. Manuscript profile -
Open Access Article
12 - Comparison of function of immobilized and free Bacillus licheniformis cells in production of alkaline protease
Mohammad Mashhadi-Karim Mehrdad Azin Seyyed Latif Mousavi Gargari Meysam SarsharAbstract Background and Objective: Proteases are an important group of industrial enzymes, which are widely used in different industries such as detergent, leather tanning, pharmaceutical, and food industries. The aim of this study was to immobilize Bacillus lichenifor MoreAbstract Background and Objective: Proteases are an important group of industrial enzymes, which are widely used in different industries such as detergent, leather tanning, pharmaceutical, and food industries. The aim of this study was to immobilize Bacillus licheniformis cells in calcium alginate beads and study of its effects on the amount of alkaline protease production. Effects of several different conditions on stability of the beads were also examined. Material and Methods: Bacillus licheniformis cells were immobilized in calcium alginate beads and were used for production of alkaline protease. The amount of enzyme production was compared in immobilized and free-cell fermentation. Effect of stuffing rate (%) on the enzyme production was studied. Optimum pH and temperature of enzyme activity were also determined. Furthermore, effects of pH, curing time and treating the beads by glutaraldehyde on stability of the beads were examined. Results: In this study the amount of production and productivity of protease in immobilized cells state showed an increase of 74% and 54% in comparison to free cells state, respectively. The highest amount of the production of the enzyme was obtained in stuffing rate of 5% (v/v). Optimum pH and temperature of the enzyme activity were determined 8 and 65oC, respectively. The highest stability of the beads was observed at curing time of 1 hour at pH of 7.4. Treating the beads by glutaraldehyde was detrimental to their stability. Conclusion: Use of immobilized cells of Bacillus licheniformis in calcium alginate beads on the one hand, can increase productivity of the alkaline protease in comparison to free cells method, and on the other hand, reduces the cost of the enzyme production because of eliminating the need of preparation of inoculum for the new batches. Manuscript profile -
Open Access Article
13 - Evaluation of Anti-inflammatory Effects of Bioactive Peptides of Spirulina Platensis Extracted by Animal Cysteine Protease Enzyme in Mice Balb/C
Samaneh Moghadamzadegan Mozhgan Emtyazjoo Mahnazsadat Sadeghi Mohammad RabbaniInflammatory bowel disease is a chronic disease of the gastrointestinal tract with no definitive drug treatment. If the disease is not prevented, it may lead to dangerous diseases such as colon cancer; thus, it is important to treat or provide medication for this diseas MoreInflammatory bowel disease is a chronic disease of the gastrointestinal tract with no definitive drug treatment. If the disease is not prevented, it may lead to dangerous diseases such as colon cancer; thus, it is important to treat or provide medication for this disease. Extensive tests performed on Spirulina show that this cyanobacterium is a unique source of natural bioactive substances with potential healing properties. The aim of this study was to prepare kiwifruit cysteine protease enzyme and optimize the extraction of the bioactive peptide Spirulina platensis by cysteine protease as well as evaluating the anti-inflammatory properties of spirulina-extracted peptides in mice with intestinal inflammation. To this end, the optimal performance of the enzyme was determined by the RSM response surface methodology and the peptides extracted in the freeze dryer were SDS page space to determine the molecular weight of the peptide by electrophoresis. The resulting peptides were stored at -20°C. To evaluate the anti-inflammatory effects in 28Balb/C mice, inflammation was induced with 4% acetic acid. After confirming the inflammation of the peptide with a concentration of 3.8 mg/kg and omeprazole as positive control and water as negative control, Balb/C mice with intestinal ulcers were gavaged. The results showed that the molecular weight of cysteine protease enzyme is 24 kDa and the optimal enzyme hydrolysis conditions of cysteine protease were obtained at 60 °C, 210 min, and pH 7.5. Histological results showed the presence of inflammation in the control group and in mice whose peptide at a concentration of 3.8 (mg/kg) had completely regenerated intestinal tissue, crypt, and intestinal mucosa during treatment. The enzyme alkaline phosphatase is low in mice receiving the peptide, indicating improved inflammation in mice. As a result, the bioactive spirulina peptides were able to treat inflammation in the intestines of mice. Manuscript profile -
Open Access Article
14 - The Effects of Phytase and Protease Supplementation in Diets Containing Seasame Meal on Nutrient Digestability, Bone Properties, Immunity and Blood Parameters in Broiler Chicken
Hesamoddin Farrokhi Rohullah Abdullahpour Vahid RezaeipourThis study was conducted to investigate the effects of supplemented phytase and protease enzymes in diets containing 15% of sesame meal on digestibility, bone physicochemical properties, immunity and blood parameters in broiler chicken. An experiment was performed in a MoreThis study was conducted to investigate the effects of supplemented phytase and protease enzymes in diets containing 15% of sesame meal on digestibility, bone physicochemical properties, immunity and blood parameters in broiler chicken. An experiment was performed in a completely randomized design using five dietary treatments with six replicates in which 300 one-day-old Ross 308 commercial broiler chicks were reared for 6 weeks. Dietary treatments were 1) corn-soybean meal based diet (negative control) 2) a corn-soybean meal diet including 15% sesame meal (positive control), 3) positive control diet supplemented with phytase, 4) positive control diet supplemented with protease and 5) positive control diet supplemented with both phytase and protease enzymes. The results indicated that ileal digestibility for crude protein was increased in chicks treated with enzymes (p < 0.05). The ileal digestibility for calcium and phosphorus were higher in chicks fed both phytase and protease (p < 0.05). Experimental treatments had no significant effects on the viscosity of gastrointestinal ileal digesta (p > 0.05). Tibial bone breaking strength in chicks treated with both enzymes increased (p < 0.05), also treatments containing phytase (with or without protease) led to more bone calcium and phosphorus (p < 0.05). Experimental treatments had no significant effect on bursa and spleen weights (p < 0.05). Blood cholestrol concentration in chicks treated with both enzymes were decreased (p < 0.05). Phytase (with or without protease) in diet, decreased blood triglyceride concentration (p < 0.05). Lower total protein was in chicks fed protease (p < 0.05). Blood calcium concentration in chicks fed enzymes were higher (p < 0.05). Supplementing broiler chicken diets with phytase and protease containing 15% sesame meal, had beneficial effects on protein, calcium and phosphorus digestibility, bone resistance, and some blood parameters. . Manuscript profile
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