Papers by Arastoo Badoei-Dalfard
Recently, hierarchical magnetic enzyme nanoflowers have been found extensive attention for effici... more Recently, hierarchical magnetic enzyme nanoflowers have been found extensive attention for efficient enzyme immobilization due to high surface area, low mass transfer limitations, active site accessibility, promotion of the enzymatic performance, and facile reusing. Herein, we report the purification of the Bacillus collagenase and then synthesis of magnetic cross-linked collagenase-metal hybrid nanoflowers (mcCNFs). The catalytic efficiency (k cat /K m) value of the immobilized collagenase was 2.2 times more than that of the free collagenase. The collagenase activity of mcCNFs enhanced about 2.9 and 4.6 at 85 and 90 • C, respectively, compared to free collagenase. Thermal stability of mcCNFs increased about 31% and 24% after 3 h of incubation at 50 and 60 • C, respectively. After 10 cycles of reusing, the mCNFs collagenase showed 83% of its initial activity. Results showed that the mcCNFs revealed 1.4 times more activity than the free collagenase in 0.16% protein waste. Furthermore, the hydrolysis value of chicken pie protein wastes by the immobilized enzyme obtained 4 times more than the free collagenase after 240 min incubation at 40 • C. Finally, our results showed that the construction of mcCNFs is an efficient method to increase the enzymatic performance and has excessive potential for the hydrolysis of protein wastes in the food industry.
Microbial alpha-amylases demonstrate more compatibility with industrial demands. These industrial... more Microbial alpha-amylases demonstrate more compatibility with industrial demands. These industrial enzymes have potential applications in various industrial parts, e.g., starch processing, brewing, baking bread, pharmaceuticals, and detergents. In this study, a mesophyll bacterium, Bacillus mojavensis strain UMF29, has been identified based on the 16S rDNA sequence and phylogenetic tree. After 72 h of incubation at 37 °C, the plates were inoculated with Lugol's iodine, and the alpha-amylase-producing isolates were distinguished by clear zones in the blue background of starch agar plate around the colonies. Biochemical characterization of this enzyme was also investigated. Results showed that the optimal activity of this enzyme was at 50 °C, and pH of 7.0. In addition, the alpha-amylase exhibited optimum stability at 40 °C, and pH of 7.0. Some metal ions including Mn 2+ , Cu 2+ , Ca 2+ , Fe 2+ , Zn 2+ , Hg 2+ , and Mg 2+ stimulated the alpha-amylase activity by about 122, 105, 61, 47, 46, 23 and 16%, respectively. The best activity of this enzyme was achieved in 0.5 M of KCl (81% enhancement), and 1.5 M of NaCl (9% enhancement). This alpha-amylase hydrolyzed a wide range of raw-starch granules (1.0 %, w/v) including potato, corn, grain, rice, and wheat and optimally was effective on wheat starch (16%, w/v) at 45 °C for 6 h with relative hydrolyses of ~10 U/ml. To our knowledge, it was also found that the alpha-amylase was a Ca 2+-dependent enzyme for hydrolyzing higher concentrations of raw wheat starch (90-180 mg/ml) after 6 h of incubation at 45 ℃. Finally, these results indicated that UMF29 alpha-amylase showed high capacity in the degradation of the raw starch.
In the present study, Ta-MOF@Fe 3 O 4 core/shell nanostructures were synthesized in optimal condi... more In the present study, Ta-MOF@Fe 3 O 4 core/shell nanostructures were synthesized in optimal conditions using the rapid, efficient, and novel ultrasound assisted reverse micelle method. FTIR, TGA/DTG, XRD, TEM, EDS and N 2 adsorption/desorption isotherms were conducted in order to obtain samples with desirable properties. Results showed that the synthesized products had the thermal stability of 200°C, particle-size distribution of 38 nm and surface area of 740 m 2 /g. Also, the VSM test showed that these compounds have desirable magnetic properties which provide the opportunity for recovery. Based on these obtained properties, final products were used as a novel candidate for enzyme immobilization. Results of SEM images revealed that the Bacillus licheniformis Km12 lipase is efficiently loaded on the Ta-MOF@Fe 3 O 4 core/shell substrate. The stability test indicated the high stability of the enzyme loaded into these nanostructures. The synthesis method and the results obtained from enzyme immobilization developed in this study can be a new strategy for various applications of these novel compounds in diverse biological fields.
In this research, lipase Km12 was immobilized on the glutaraldehyde-activated graphene oxide/chit... more In this research, lipase Km12 was immobilized on the glutaraldehyde-activated graphene oxide/chitosan/cellulose acetate nanofibers (GO/Chit/CA NFs) prepared by the electrospinning method. This immobilized lipase exhibited a higher activity value than the free lipase in the acidic pH region. This enzyme showed a 10 • C shift in the maximum temperature activity. Results displayed that the Vmax value of NFs-lipase was 0.64 µmol/min, while it was gained 0.405 µmol/min for the free lipase. The activity of NFs-lipase was reserved 100% after 10 min maintaining at 60 • C, in which the free lipase only kept 75% of its original activity. Moreover, a 20% enhancement in the lipase activity was observed for NFs-lipase after 180 min of incubation at 60 • C, compared to the free enzyme. Reusability studies exhibited that the immobilized lipase well-kept 80% of its original activity after 10 cycles of reusing. Results displayed that 14% of the protein was leaked from NFs-lipase at the same condition. Transesterification results indicated that the free lipase exhibited 65% and 85% conversation level of benzyl acetate after 12 and 24 h of incubation. Besides, the immobilized lipase showed 80% and 95% conversation level at the same condition. These results indicated the high performance of free and immobilized lipase in the production of benzyl acetate for applications in the perfume and cosmetic industries.
Recently, biodiesel production has received extensive attention as an alternative energy source t... more Recently, biodiesel production has received extensive attention as an alternative energy source to conventional fuel. It is non-polluting fuel and combines environmental friendliness with low toxicity, renewability, and biodegradability. Immobilized lipases have been found emerging catalysts for the synthesis of biodiesel. In the current study, we used lipase Km12 to construct three efficient immobilized enzymes, including lipase@ZIF-8 (LZIF),
Phosphorus is one of the most important nutrients for plant growth and development. Chemical Pi f... more Phosphorus is one of the most important nutrients for plant growth and development. Chemical Pi fertilizer is used to provide the phosphorus for the plants, but it is mostly fixed in the soil into insoluble form and become unavailable to the plants. Phosphate-solubilizing bacteria have lots of application in agriculture as biological fertilizer. Consumption of biofertilizers instead of chemical fertilizers can lead to environmental pollution reduction and crop production enhancement using sustainable farming. In this study, a phosphatase-producing bacterium was isolated from agricultural soil in Kerman. Screening of phosphate solubilizing bacteria was performed on the PVK medium, based on clear area diameter. The best bacterium (AG41) was identified based on 16s rDNA gene. The optimum condition for production of phosphatase was also determined and it was purified and characterized. Sequence alignment and phylogenetic tree results show that AG41 is closely related to Bacillus subtilis, with 98% homology. Phosphatase activity was determined by end point method. The best carbon, nitrogen and phosphate sources for enzyme production were 1.0% glucose, 0.5% ammonium sulfate and (0.25%) sodium phytate +(0.25%) tricalcium phosphate, respectively. Bacterial phosphatase was partially purified using ammonium sulfate fractionation followed by dialysis. Results showed that the optimum temperature for the purified enzyme activity was 40 o C and it was stable at temperatures below 60°C. This enzyme was stable between pH 3.0-7.0, and the optimal pH activity was found to 5.0. These results indicated that this strain can be a notable candidate for using as biofertilizers.
A thermophilic strain AMF-07, hydrolyzing carboxymethylcellulose (CMC) was isolated from Kerman h... more A thermophilic strain AMF-07, hydrolyzing carboxymethylcellulose (CMC) was isolated from Kerman hot spring and was identified as Bacillus licheniformis based on 16S rRNA sequence homology. The carboxymethylcellulase (CMCase) enzyme produced by the B. licheniformis was purified by (NH4)2SO4 precipitation, ion exchange and gel filtration chromatography. The purified enzyme gave a single band on SDS-PAGE with a molecular weight of 37 kDa. The CMCase enzyme was highly active and stable over broad ranges of temperature (40-80 º C), pH (6.0-10.0) and NaCl concentration (10-25%) with an optimum at 70 º C, pH 9.0 and 20% NaCl, which showed excellent thermostable, alkali-stable and halostable properties. Moreover, it displayed high activity in the presence of cyclohexane (134%) and chloroform (120%). Saccharification of rice bran and wheat bran by the CMCase enzyme resulted in respective yields of 24 and 32 g L-1 reducing sugars. The enzymatic hydrolysates of rice bran were then used as the substrate for ethanol production by Saccharomyces cerevisiae. Fermentation of cellulosic hydrolysate using S. cerevisiae, reached maximum ethanol production about 0.125 g g-1 dry substrate (pretreated wheat bran). Thus, the purified cellulase from B. licheniformis AMF-07 utilizing lignocellulosic biomass could be greatly useful to develop industrial processes.
PVA/ZnO nanofibrous composite polymer was synthesized as a novel sorbent via electrospinning meth... more PVA/ZnO nanofibrous composite polymer was synthesized as a novel sorbent via electrospinning method at ambient conditions. Physicochemical characteristics of the samples were analyzed using XRD, SEM, EDS, TGA/DSC, and BET techniques. Efficiency of the composite samples for arsenic removal [i.e., As (III) and As (V) anions] was studied under different conditions using RSM optimization approach. Experimental results of adsorption tests indicated that the synthesized PVA/ZnO polymer with the maximum removal of 97% is a highly efficient sorbent for arsenic comparable with other sorbents studied for arsenic removal. The effect of some cations on arsenic removal was also studied in this research. Results showed that the tested ions do not cause a significant reduction in the removal rate. PVA/ ZnO composite polymer, which is highly biocompatible, can be considered as a new adsorbent for removal of arsenic from contaminated water.
The acidophilic and thermophilic cellulase would facilitate the conversion of lignocellulosic bio... more The acidophilic and thermophilic cellulase would facilitate the conversion of lignocellulosic biomass to biofuel. In this study, Bacillus sonorensis HSC7 isolated as the best thermophilic cellulose degrading bacterium from Gorooh hot spring. 16S rRNA gene sequencing showed that, this strain closely related to the B. sonorensis. CMCase production was considered under varying environmental parameters. Results showed that, sucrose and (NH 4) 2 SO 4 were obtained as the best carbon and nitrogen sources for CMCase production. B. sonorensis HSC7 produced CMCase during the growth in optimized medium supplemented with agricultural wastes as sole carbon sources. The enzyme was active with optimum temperature of 70°C and the optimum CMCase activity and stability observed at pH 4.0 and 5.0, respectively. These are characteristics indicating that, this enzyme could be an acidophilic and thermophilic CMCase. Furthermore, the CMCase activity improved by methanol (166%), chloroform (152%), while it was inhibited by DMF (61%). The CMCase activity was enhanced in the presence of Mg +2 (110%), Cu +2 (116%), Triton X-100 (118%) and it retained 57% of its activity at 30% NaCl. The compatibility of HSC7 CMCase varied for each laundry detergent, with higher stability being observed in the presence of TajÒ and daryaÒ. This enzyme, that is able to work under extreme conditions, has potential applications in various industries.
Background & Objectives: Thermophilic alpha-amylase can be used in different industries such as s... more Background & Objectives: Thermophilic alpha-amylase can be used in different industries such as starch processing and detergents. This study was performed to isolate alpha-amylase-producing bacteria and characterization of the enzyme. Materials & Methods: After sample collection from Gorooh hot spring in Kerman province, Iran, thermophilic alpha-amylase-producing bacteria were isolated using the starch-agar medium. 16S rDNA sequencing was used to identify the bacterial strain. Characterization of the thermophilic alpha-amylase was performed in the presence of various factors such as pH, temperature, metal ions, chemical compounds, and organic solvents. Also, kinetic parameters of the enzyme were determined in different concentrations of starch. Results: Anoxybacillus gonensis AT23 was identified as the best thermophilic alpha-amylaseproducing strain. The alpha-amylase enzyme showed the optimal activity at pH 5 to 6. Sevenfold increase in the enzyme activity was observed in the presence of NaCl (3M). Mn 2+ and Zn 2+ increased the enzyme activity about 95% and 31%, respectively. Kinetic parameters including K m and V max were estimated about 1.657 mg/ml and 0.0059 mg/ml/min, respectively. Also, enzyme activity was also improved about 2 folds in the presence of organic solvents including n-butanol and 10% cyclohexane. Conclusion: Our results indicated that AT23 alpha-amylase is a halophile and organic solvent-tolerant enzyme. Therefore, it can be used in different industries.
Thermo-tolerant phytase has lots of applications in diferent industries, such as environmental pr... more Thermo-tolerant phytase has lots of applications in diferent industries, such as environmental protection, aquaculture, and
agriculture. In the present study, Bacillus sp. Dm018 has been isolated from Demand hot spring in Jiroft, which placed in
the southwest of Iran. Statistical optimization approach by response surface methodology (RSM) leads to 2.3-fold growth in
phytase construction by Bacillus sp. Dm018. The optimal temperature for phytase function was established to be about 60 °C.
Furthermore the enzyme reserved more than 70% of its activity over an extensive pH series of 4.0–8. 0. Results showed that
Dm018 phytase activity was inhibited by CaCl2 (5 mM) about 60%. Phytase activity was inhibited by SDS (5 mM) about
30% and slightly improved by EDTA about 6.0%. Phytase stability data exhibited that the enzyme reserved more than 50%
of it,s original activity after pre-incubated in the existence of diverse ions, except CaCl2. These results indicate that Dm018
phytase has good latent for commercial attention as an animal feed preservative
Phytase can improve the nutritional value of plant-based foods by enhancing protein digestibility... more Phytase can improve the nutritional value of plant-based foods by enhancing protein digestibility and mineral
availability through phytate digestion in the stomach and the food processing industry. Microbial sources are more
promising for the production of phytases on a commercial scale. The objectives of this exploration were to screening
and isolation of phytase-producing bacteria from hot spring with commercial interest. Molecular identification of the
best isolate was achieved by the 16S rDNA gene. Optimization of phytase production was prepared in the presence of
different phosphate, nitrogen, and carbon sources. Enzyme activity and stability were also explored in the presence of
different pHs, temperatures, and ion compounds. Comparing the 16S rDNA gene sequence of the isolate LOR10 with
those in GenBank using Clustal omega shows 98% sequence homology with Bacillus amyloliquefaciens. Medium
optimization studies showed that galactose, yeast extract, and tricalcium phosphate were the best sources of carbon,
nitrogen, and phosphate for phytase production, respectively. The optimum temperature activity was also observed to be
70 oC. Phytase stability was at its optimum in a pH range of 5.0–8.0. Phytase activity increased in the presence of CaCl2,
ZnCl2, and MnSO4 about 1.4, 2.3 and 1.6 folds, respectively. It could be mentioned that phytase activity decreased by
about 30 % in the presence of EDTA and SDS. On the basis of the results, it could be concluded that LOR10 phytase
has a great potential for commercial interest as an additive to animal plant-based foods.
Aromatic hydrocarbons are carcinogenic contaminants produced through natural and anthropogenic so... more Aromatic hydrocarbons are carcinogenic contaminants produced through natural and anthropogenic sources. The utilization of bacteria plays a crucial role in aromatic compounds bioremediation. For the isolation of mono-and di-aromatic hydrocarbon (phenol and naphthalene) degrading bacteria, samples were collected from five zones in Sarcheshmeh copper mine, Iran.
Naphthalene is an ubiquitous pollutant of the environment and the biodegradation of this pollutan... more Naphthalene is an ubiquitous pollutant of the environment and the biodegradation of this pollutant has been
receiving constant scientific consideration. The aim of this study was to isolate and identify bacteria that could
degrade naphthalene from three regions of the Gol Gohar Mine at Sirjan, Iran. In this study, the total naphthalene
degrading bacteria were quantified with the most probable number (MPN) and the colony forming unit (CFU)
methods. The results showed that most of the bacteria communities capable of degrading naphthalene aggregated in
the (WG) site. Among 22 isolated bacteria, seven strains were selected for their ability to grow at higher
concentrations of naphthalene (300 and 400 mg/l) and biochemical characteristics. Finally, two strains named
isolates 72N and 79N were selected for analysis of the 16S rRNA sequences. Strain 72N was identified as
Pseudomonas fluorescens AHB72N and strain 79N was shown to be related to Pseudomonas gessardii AHB79N.
The results of biodegradation tests showed that these two strains could degrade 600 mg/l naphthalene in 7 days. The
results indicated that strain 79N showed higher potential for removing naphthalene than strain 72N. Practical
application of bacterial strains for the degradation of naphthalene from the industrial zones opens interesting
prospects. The results of this study provide useful information in evaluating naphthalene degraders isolated from
wastewater and industrial sites
PAHs are aromatic hydrocarbons with two or more fused benzene rings. They are formed during the t... more PAHs are aromatic hydrocarbons with two or more fused benzene rings. They are formed during the thermal breakdown of organic molecules and their succeeding recombination. Naphthalene is the simplest (PAHs) that is carcinogenic. Bioremediation method is considered as an economical and safe approach for the elimination of aromatic compounds from environment. The bacteria were capable to grow on various hydrocarbons like naphthalene. The aim of this research is to isolate and identify naphthalene-degrading bacteria from the coal mine of Zarand. Four samples of water and sludge from various sites of the mine were collected. These sites include the following: Main coal vacate site (MC), Inoculum Sump site (IB), Sludge aggregate site (SA), and Near sludge aggregate site (NF). In this study, 12 bacterial strains that utilize naphthalene at initial concentration 200 mg/L (ppm) as carbon and energy sources for growth were isolated from the Zarand mine in Iran. In addition, bacterial cell density was assayed by measuring the OD 600. In addition, total naphthalenedegrading bacteria were quantified with the most probable number (MPN) procedure using microtiter plates and the colony-forming unit (CFU) method. The results had shown that most of the naphthalene degrader bacteria aggregated in (SA) site. Six bacteria, isolated from wastewater and oil-contaminated soil showed great potential as naphthalene degraders up to 400 (ppm) and selected for biochemical characteristics. Naphthalene tolerance of the strains in various concentrations of naphthalene indicates that the strain 38 N can grow best at 600 (ppm) naphthalene. This strain was identified based on 16S rRNA gene analysis that showed belonging to Sphingobacterium multivorum AHB38N.
Phenol is one of the major aromatic pollutants among variety of toxic compounds. Phenol is a man-... more Phenol is one of the major aromatic pollutants among variety of toxic compounds. Phenol is a man-made as well as a naturally occurring aromatic compound and an important intermediate in the biodegradation of natural and industrial aromatic compounds. To clean up the aromatic contaminated sites a bioremediation method is considered as an economical and safe approach for the environment. In this study, 10 phenol-degrading bacterial strains were isolated from three sites at Midok copper mine in Shahrbabak. These sites include: Magnetit site (WG), Hematit site (WH) and Near Hematit site (NH). Total heterotrophic and phenol degrading bacteria were quantified with most probable number (MPN) and colony forming unit (CFU) methods. The results of this study show that the (WG) site of the mine have the highest phenol degrading bacteria. Totally 10 phenol degrading bacteria were isolated from three sites in Midok copper mine. Then, five bacterial strains were selected according to high growth rate and phenol degradation. Finally two strains named isolates P62 and 69P were selected for analysis of 16S rRNA sequences. Strain P62 belongs to Pseudomonas putidaAHBP62 and strain P69 is related to Arthrobacter scleromaeAHB69P that has capability degradation of 600 (ppm) phenols in 7 days. By using these degradative bacteria in contaminated mine sites the environmental pollution can be managed.
Organic solvents tend to strip water from protein and thereby disrupt non-covalent forces and dec... more Organic solvents tend to strip water from protein and thereby disrupt non-covalent forces and decrease enzyme activity and stability. In the present study, we have replaced the surface charge residues in Salinivibrio zincmetalloprotease (SVP) with hydrophobic ones (E12V, D22I, D24A and D310I) in order to study the effects of surface hydrophobicity with hydrophobic strength of organic solvents. Compared to SVP, D24A exhibited an increase in k cat and catalytic efficiency and a reduction in thermal inactivation rate in aqueous solvent. Structural studies indicated that the replacement of surface charge residues with hydrophobic residues would not induce conformational changes. C 50 value (the value of solvent concentration where 50% of enzyme activity remains), k i (irreversible thermoinactivation rate), and kinetic parameters of E12V, D22I, and D24A were higher in isopropanol and n-propanol. D24A is found to be the most efficient mutant for its remarkable decrease in k i value in the presence of isopropanol and n-propanol and a reduction in k i value in the presence of dimethylformamide (DMF) and methanol. C 50 value in this variant was increased about 1.2% in DMF, 2% in methanol and isopropanol and 2.5% in n-propanol. Results revealed that, there was a correlation between surface hydrophobicity of SVP and hydrophobic strength of organic solvents.
Recently, enzyme immobilization via self-assembly of magnetic metal-organic frameworks (mMOFs) ha... more Recently, enzyme immobilization via self-assembly of magnetic metal-organic frameworks (mMOFs) has attracted attention for researchers. In this study, a Bacillus protease KHB3 was embedded into a magnetic metal organic framework. Temperature activity results shown that the free protease displayed the optimal activity at 40 °C, while the immobilized protease showed the highest activity at 60 °C. Besides, a 20% improvement in the activity of the immobilized form was found at 80 °C against the free protease. Furthermore, the Vmax of immobilized enzyme was 1.75 fold more than the free enzyme. Results exhibited that the immobilized protease showed 1.54 and 1.55 folds more activity than the control toward fibrin and gelatin, respectively. The immobilized protease reserved about twofold higher activity than the free protease after 21 days of storage. Results revealed that the immobilized enzyme showed 38% clot lysis, while the free protease showed 26% at a similar condition. The hydrolysis degree of the free protease and immobilized protease was obtained about 17 and 45% after 8 h incubation at 50 °C, respectively. Overall, our results show the high potential of these enzymes in the clot lysis capacity and construction of high-value compounds from protein hydrolysis of shrimp protein waste.
Due to the increasing development of the poultry industry and increasing environmental pollution,... more Due to the increasing development of the poultry industry and increasing environmental pollution, especially keratin waste, efficient keratin hydrolyzing biocatalyst has become an attractive goal for researchers. In the current research, a Bacillus keratinase has been immobilized based on the cross-linked enzyme aggregates (CLEAs) method on the functionalized Fe 3 O 4 nanoparticles. Immobilization efficiency and the relative activity of the immobilized enzyme were 87 and 84%, respectively. Results exhibited that the immobilized keratinase showed 7.96 folds more Vmax value than free enzyme in the presence of keratin substrate. Thermostability results showed that the mCLEAs-keratinase improved the enzyme stabilities by about 3.5 and 5.8 folds compared to free enzyme after 3 h incubation at 70 and 80 °C, respectively. The substrate specificity results showed that the activity of keratinase 2S1 toward keratin was five times more than the casein substrate. The activity of the immobilized enzyme in the presence of acetonitrile increased about 7.3 times compared to the free keratinase. The results showed that 43% of the enzyme activity was maintained after ten cycles of reusing. The hydrolysis degree of keratin wastes by immobilized enzyme was three folds more than free enzyme after 2 h of incubation at 42 °C. These results indicated that the free and immobilized keratinases had high potential activities for recycling keratin waste.
Protease is one of the most important industrial enzymes occupying nearly 60% of global enzyme sa... more Protease 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.
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Papers by Arastoo Badoei-Dalfard
agriculture. In the present study, Bacillus sp. Dm018 has been isolated from Demand hot spring in Jiroft, which placed in
the southwest of Iran. Statistical optimization approach by response surface methodology (RSM) leads to 2.3-fold growth in
phytase construction by Bacillus sp. Dm018. The optimal temperature for phytase function was established to be about 60 °C.
Furthermore the enzyme reserved more than 70% of its activity over an extensive pH series of 4.0–8. 0. Results showed that
Dm018 phytase activity was inhibited by CaCl2 (5 mM) about 60%. Phytase activity was inhibited by SDS (5 mM) about
30% and slightly improved by EDTA about 6.0%. Phytase stability data exhibited that the enzyme reserved more than 50%
of it,s original activity after pre-incubated in the existence of diverse ions, except CaCl2. These results indicate that Dm018
phytase has good latent for commercial attention as an animal feed preservative
availability through phytate digestion in the stomach and the food processing industry. Microbial sources are more
promising for the production of phytases on a commercial scale. The objectives of this exploration were to screening
and isolation of phytase-producing bacteria from hot spring with commercial interest. Molecular identification of the
best isolate was achieved by the 16S rDNA gene. Optimization of phytase production was prepared in the presence of
different phosphate, nitrogen, and carbon sources. Enzyme activity and stability were also explored in the presence of
different pHs, temperatures, and ion compounds. Comparing the 16S rDNA gene sequence of the isolate LOR10 with
those in GenBank using Clustal omega shows 98% sequence homology with Bacillus amyloliquefaciens. Medium
optimization studies showed that galactose, yeast extract, and tricalcium phosphate were the best sources of carbon,
nitrogen, and phosphate for phytase production, respectively. The optimum temperature activity was also observed to be
70 oC. Phytase stability was at its optimum in a pH range of 5.0–8.0. Phytase activity increased in the presence of CaCl2,
ZnCl2, and MnSO4 about 1.4, 2.3 and 1.6 folds, respectively. It could be mentioned that phytase activity decreased by
about 30 % in the presence of EDTA and SDS. On the basis of the results, it could be concluded that LOR10 phytase
has a great potential for commercial interest as an additive to animal plant-based foods.
receiving constant scientific consideration. The aim of this study was to isolate and identify bacteria that could
degrade naphthalene from three regions of the Gol Gohar Mine at Sirjan, Iran. In this study, the total naphthalene
degrading bacteria were quantified with the most probable number (MPN) and the colony forming unit (CFU)
methods. The results showed that most of the bacteria communities capable of degrading naphthalene aggregated in
the (WG) site. Among 22 isolated bacteria, seven strains were selected for their ability to grow at higher
concentrations of naphthalene (300 and 400 mg/l) and biochemical characteristics. Finally, two strains named
isolates 72N and 79N were selected for analysis of the 16S rRNA sequences. Strain 72N was identified as
Pseudomonas fluorescens AHB72N and strain 79N was shown to be related to Pseudomonas gessardii AHB79N.
The results of biodegradation tests showed that these two strains could degrade 600 mg/l naphthalene in 7 days. The
results indicated that strain 79N showed higher potential for removing naphthalene than strain 72N. Practical
application of bacterial strains for the degradation of naphthalene from the industrial zones opens interesting
prospects. The results of this study provide useful information in evaluating naphthalene degraders isolated from
wastewater and industrial sites
agriculture. In the present study, Bacillus sp. Dm018 has been isolated from Demand hot spring in Jiroft, which placed in
the southwest of Iran. Statistical optimization approach by response surface methodology (RSM) leads to 2.3-fold growth in
phytase construction by Bacillus sp. Dm018. The optimal temperature for phytase function was established to be about 60 °C.
Furthermore the enzyme reserved more than 70% of its activity over an extensive pH series of 4.0–8. 0. Results showed that
Dm018 phytase activity was inhibited by CaCl2 (5 mM) about 60%. Phytase activity was inhibited by SDS (5 mM) about
30% and slightly improved by EDTA about 6.0%. Phytase stability data exhibited that the enzyme reserved more than 50%
of it,s original activity after pre-incubated in the existence of diverse ions, except CaCl2. These results indicate that Dm018
phytase has good latent for commercial attention as an animal feed preservative
availability through phytate digestion in the stomach and the food processing industry. Microbial sources are more
promising for the production of phytases on a commercial scale. The objectives of this exploration were to screening
and isolation of phytase-producing bacteria from hot spring with commercial interest. Molecular identification of the
best isolate was achieved by the 16S rDNA gene. Optimization of phytase production was prepared in the presence of
different phosphate, nitrogen, and carbon sources. Enzyme activity and stability were also explored in the presence of
different pHs, temperatures, and ion compounds. Comparing the 16S rDNA gene sequence of the isolate LOR10 with
those in GenBank using Clustal omega shows 98% sequence homology with Bacillus amyloliquefaciens. Medium
optimization studies showed that galactose, yeast extract, and tricalcium phosphate were the best sources of carbon,
nitrogen, and phosphate for phytase production, respectively. The optimum temperature activity was also observed to be
70 oC. Phytase stability was at its optimum in a pH range of 5.0–8.0. Phytase activity increased in the presence of CaCl2,
ZnCl2, and MnSO4 about 1.4, 2.3 and 1.6 folds, respectively. It could be mentioned that phytase activity decreased by
about 30 % in the presence of EDTA and SDS. On the basis of the results, it could be concluded that LOR10 phytase
has a great potential for commercial interest as an additive to animal plant-based foods.
receiving constant scientific consideration. The aim of this study was to isolate and identify bacteria that could
degrade naphthalene from three regions of the Gol Gohar Mine at Sirjan, Iran. In this study, the total naphthalene
degrading bacteria were quantified with the most probable number (MPN) and the colony forming unit (CFU)
methods. The results showed that most of the bacteria communities capable of degrading naphthalene aggregated in
the (WG) site. Among 22 isolated bacteria, seven strains were selected for their ability to grow at higher
concentrations of naphthalene (300 and 400 mg/l) and biochemical characteristics. Finally, two strains named
isolates 72N and 79N were selected for analysis of the 16S rRNA sequences. Strain 72N was identified as
Pseudomonas fluorescens AHB72N and strain 79N was shown to be related to Pseudomonas gessardii AHB79N.
The results of biodegradation tests showed that these two strains could degrade 600 mg/l naphthalene in 7 days. The
results indicated that strain 79N showed higher potential for removing naphthalene than strain 72N. Practical
application of bacterial strains for the degradation of naphthalene from the industrial zones opens interesting
prospects. The results of this study provide useful information in evaluating naphthalene degraders isolated from
wastewater and industrial sites