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Tomato is attributed as a global host for root-knot nematode (Meloidogyne incognita) soliciting ponderous damage. Using biocontrol agents to control plant parasitic nematodes is a well-established, green approach in advance of synthetic nematicides. The role of Bacillus spp. in inciting physiological and biochemical alterations in nematode infestation is discussed in the present study. The susceptible (PKM-1) and resistant (Hisar Lalit) tomato cultivars treated with Bacillus pumilus augmented the shoot length, root length and biomass of plants compared to the standard check, Pseudomonas fluorescens, followed by B. megaterium. Accordingly, all the biocontrol agent-treated susceptible plants showed reduced galling and exhibited a root gall index of 3 (moderately resistant). Contrarily, all the resistant plants showed highly resistant reactions. B. pumilus showed the topmost expression of all the biochemical enzymes like peroxidase (PO), polyphenol oxidase (PPO), catalase (CAT), phenylalanine ammonia-lyase (PAL) and total phenols. Conclusively, B. pumilus was found to be the most potential in reducing nematode infestation by embellishing the plant growth and enhancing defense-related enzymes in tomatoes.

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The tomato leaf miner, Tuta absoluta (Meyrick, 1917) (Lepidoptera: Gelechiidae), is one of the most important pests causing significant economic losses in plant species belongingthe Solanaceae family. The preferred management method for T. absoluta currently involves insecticide application. However, beside the undesired effects of insecticides, chemical treatments can also negatively impact the efficiency of integrated pest management programs (IPM). Bacillus thuringiensis (Shigetane 1902) (Bacillales: Bacillaceae) (Bt) is a pathogen with formulations used as host-specific bioinsecticides. These formulations decompose quickly in the environment, thereby reducing non-target effects and residue problems compared to chemical pesticides. In this study, the effectiveness of six commercial Bt products, belonging to aizawai and kurstaki strains, against T. absoluta was assessed under laboratory conditions, using manufacturer-recommended doses. The efficacy of the Bt products varied between 70 and 97.5%. The lowest and highest mortalities were recorded in B. thuringiensis var. aizawai and B. thuringiensis var. kurstaki products, respectively. Mortality reached 100% within three days following insecticide treatments, whereas peak mortality in Bt applications was noted after a post-treatment period of fifteen days. These findings highlight the potential of certain Bt products as effective components of IPM programs for T. absoluta, suggesting the need for further field studies to optimize their use in agricultural practices.

 

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Background: Most of the population in the different areas of world is affected by bacterial infections responsible for dental caries. Due to the importance of traditional medicines derived from herbs used for dental problems, this study investigated in vitro antibacterial activity of Mentha longifolia essential oil from Ardabil, Iran, on Streptococcus mutans, Lactobacillus rhamnosus and Actinomyces viscosus,bacteria that cause tooth decay. Materials and Methods: The volatile oil of Mentha longifolia leaves was extracted by hydrodistillation method using a Clevenger-type apparatus  and analyzed by GC and GC/MS system.The antibacterial activity was evaluated by the disk diffusion susceptibility in dilutions of 62.5, 125, 250, 500, 1000 and 2000μg/μl and broth macrodilution test methods. Results: The oil was particularly rich in Pulegone (31.78%), 1,8-cineole (15.99%), menthoforan (11.25%), cis-isopulegon (10.5%) and paramenth-3-n-8-l (6.85%). The medicinal plant essential oil could prevent the growth of the bacteria, and the rates of MIC and MBC of native pennyroyal essential oil on Streptococcus mutans, Lactobacillus rhamnosus and Actinomyces viscosus were 110, 165, 80, 120, 450 and 650μg/μl, respectively. The maximum inhibition zone diameter was about 12.2, 27.2 and 4.8 mm, for Streptococcus mutans, Lactobacillus rhamnosus and Actinomyces viscosus respectively, at the concentration of 500μgμl^-1. Conclusion: In this work, the essential oil of medicinal plant containing effective ingredients could prevent the growth of bacteria and may be used as an affordable and available source for medicinal purposes.

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Electronic wastes are known as the most important solid wastes in 21th century. They are producing two or three time faster than other solid waste streams. Many researchers studied bioleaching of e-wastes using Acidithiobacillus ferrooxidans. The presence of alkaline metals cause e-wastes show an alkalinity nature. By adding e-wastes to the environment the pH of solution increases sensibly. Many researchers supposed the optimal pH range of A. ferrooxidans which is 1.5-2.5 as the optimal pH range to reach maximum recovery. So in the bioleaching process by daily pH adjusting in the range and using sensible amount of sulfuric acid, control the pH of solution about 2. In this research two same experiments, just the pH of one of them was adjusted daily, were done. In both experiments the environmental situation including pulp density of 15 g/l, inoculum 10% (v/v), the temperature of 30ºC, and shaking rate of 130 rpm was the same. For 25 days Cu recovery, bacterial count, pH, and Eh were examined. The results showed the maximum Cu recovery at the sample without pH adjusting was about 100% but at the sample with pH adjusting recovery was reduced to 90%. The bacterial count diagram showed the bacterium is well active in both experiments. To maximize recovery, reducing acid consumption, and increasing process economy there is no need to adjust the pH of solution.

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Soybean, Glycine max, is susceptible to a large number of disease agents such as seedling and root pathogens that cause serious damages to this crop plant. One of these soil-borne pathogens is Macrophomina phaseolina, the causal agent of charcoal root rot. In this study, two bacteria, Pantoea agglomerans and Bacillus sp. BIN, and a fungus, Trichoderma harzianum T100, as potential biocontrol agents, and maneb fungicide, were evaluated against soybean charcoal rot disease in In Vitro and greenhouse conditions. All antagonists inhibited growth of the pathogen in dual culture test by 73.8, 63.3 and 55.3 %, respectively. Data from greenhouse experiments showed that in the presence of pathogen all antagonists increased the growth indices of soybean in both pasteurized and non-pasteurized soil. Reductions of microsclerotia coverage on soybean root and stem by P. agglomerans, Bacillus sp. and T. harzianum were up to 62.5, 87.6 and 62.5 %, respectively and for maneb fungicide was 87.6 % in pasteurized soil. The overall results of this study show high capability of used antagonists in reduction of initial inoculums for next season of this monocyclic disease.

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Probiotics are live microbial feed supplements, which beneficially affect the host by improving its intestinal microbial balance[8].Recently, researches have shown that lactic Acid Bacteria(LAB), especially Bacillus (because of having endospore) remain stable at cooking temperature and retain their probiotic benefits in baked goods [3]. The purposeof this study,was using of Bacillus coagolans, as a resistant probiotic,for enrichment of  bread. First, these probiotic bacteria were determined by the tests of salt tolerance, heat resistance, bile tolerance, tolerance of acid and pepcin, resistance of antibiotics, and preventing the growth of pathogenic starins[18].Then a certain number cells of  BC were entered in bread dough,before& after  baking the number of live bacteria were calculated by colony count in 1g of dough and bread. The number of BC decreased from 10⁸ to 10⁶ units per gram, after baking. Also,the amount of starch decreasd and changed into simple sugers. The pH was estimated about 4.5-5 and TTA (Total Titritable Acidity) was between 6-8. Finally, the enrichment of bread was  evaluated  by experts and its quality and taste were compared with a control sample. The Results showed this bacterium survives in baked bread and makes good chemical changes on it.

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In the  Bacillusamyloliquefaciens α-amylase (BAA), the loop (residues from 177-185; region І) is the constructive part of the cage responsible for attachment to calcium. It has two more amino acid residues than the α-amylase from Bacillus licheniformis (BLA). Arg176 in this region makes an ionic interaction with Glu126 from region ІІ (residues 118-131) but this interaction is lost in BLA due to substitution of R176Q and E126V. It is the common feature of α-amylases that calcium ion is required for their thermal stability. The present work quantitatively estimates the effect of ionic interaction on the overall stability of the enzyme. To clarify the functional and structural significance of corresponding salt bridge, first an automated homology model of the mutant enzyme (∆E126) was built by the Swiss-Model Protein Modeling Server.  Bacillus amyloliquefaciens α-amylase (3BH4.pdb) was used as the template and examined by GETAREA and WHAT IF programs, then Glu126 was deleted (∆E126) by site-directed mutagenesis and the thermostability was examined for the wild-type and mutant enzymes. Modeling results showed that deletion of salt bridge affected on the hydrophobic and hydrophilic residues orientation of two discussed regions (Ι, ΙΙ). The mutant enzyme also exhibited lower thermostability relative to the wild-type enzyme. Thus, it may be suggested that salt bridge could affect on accessible surface area of the discussed regions, decrease water diffusion,  prevent diffusion of cations and improve the thermostability of the whole protein.

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Background: Recently, the use of probiotics in preventing and treating the immune system diseases through changes in blood factors has attracted the attention of researchers. Therefore, the aim of this study was to evaluate the effect of Lactobacillus plantarum and Bifidobacterium B94 on changes of blood factors, influencing the autoimmune system diseases.
Materials and Methods: The rats used in this study were divided into four groups (n=10 each), including control (saline), damage with Ethidium bromide (EB), L. plantarum and Bifidobacterium B94 treatment groups. In damage and treatment groups, a single dose of 3μL EB was directly injected into hippocampus of rats for inducing demyelization. Also, in control group, the same amount of saline was used. Then 2×10⁸ probiotic bacteria were administered by gavage for 28 days. Then serum calcium and cholesterol levels were measured. Data were analyzed by one-way ANOVA and Tukey post-hoc tests (p≤ .05).
Results: The results showed that level of blood serum calcium increased insignificantly in the L. plantarum and Bifidobacterium B94 treatment groups compared to control group.  Also, the level of blood serum cholesterol decreased insignificantly in both treatment groups compared to control group.
Conclusion: Probiotics are used for preventing and treating some of the common autoimmune diseases such as MS. Previous studies showed that probiotics affects some of the blood parameters such as calcium and cholesterol while decrease or increase in these parameters is effective in the improvement of MS.  Although no significant finding has been obtained in some of these studies, they have almost confirmed the recommendation of probiotic consumption.

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Abstract: ATP sulfurylase (ATPS) is widely distributed in all living organisms. Several different physiological roles have been proposed for ATPS in different species, including sulfate assimilation, sulfate reduction and pyrophosphate recycling. Also, ATP sulfurylase has many different industrial and laboratory applications. The aim of this study was to clone and express the gene that producing the recombinant ATPS protein from an Iranian strain of Geobacillus. After Isolation and identification of Geobacillus kaustophilus strain, DNA genomic was extracted. ATPS gene was amplified from genomic DNA by using a couple of specific primers for interested gene. PCR product of ATPS gene was observed as an 1188bp band on agarose gel. Then the PCR product was purified and cloned into the cloning vector. The ATPS band was sequenced after cloning and result of homology search in the NCBI database confirmed that the cloned gene was ATPS. The ATPS gene was subcloned in expression pET28a plasmid. Expression of recombinant ATPS protein in E. Coli BL21 (DE3) was analyzed using SDS-PAGE gel. Analysis of expressed ATPS protein on SDS-PAGE gel revealed a band at 47.5 KD. Using ATP luminescence method for measuring enzymatic activity of the protein showed that the recombinant protein is active. This is the first study on cloning, expression and enzymatic activity of the ATPS gene from the Geobacillus kaustophilus bacteria.

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The growth performance of cyprinids larvae (Cyprinuscarpio,Hypophthalmichthysnobilisand Ctenopharyngodonidella) fed with the nauplii fromdifferentArtemiaspecies, including A. parthenogenetica, A. fransiscanaand A. urmiana as well as a probiotic mixture (Bacillus subtilis and B. licheniformis) was investigated. Fish larvae (100±15 mg) were allocated into 54 circular fiberglass tanks filled with 10 liters of water (density of 5 fish per liter) and fed for 4 weeks with the designated diets. At the end of the feeding trial, feed and growth indices [final weight, specific growth rate (SGR), condition factor (CF) and food efficiency] and body composition (crude protein, crude lipid, ash and moisture) were assessed. Significant differences were observed in feed and growth parameters especially in terms of specific growth rate and feed efficiency (P< 0.05). The highest and lowest growth and feeding efficiency were observed in C. idella fed with A. parthenogeneticanauplii and H.nobilis fed with A. urmiananauplii,respectively. Subsequently, elevated growth performance (final weight, final length and SGR) was observed in C. idella fed with nauplii of A. parthenogenetica and probiotic compared to other group (P < 0.05).

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The present study was conducted to evaluate the effect of Lactobacillus plantarum (KC426951) isolated from the intestine of rainbow trout Guilan on growth factors, carcass composition and the intestinal bacterial flora of rainbow trout (Oncorhynchus mykiss) was conducted. Rainbow trout weighing 3.56±2.24 for 2 weeks were consistent with environmental rearing conditions. Five groups of fish were fed with diets containing 10⁶ (T1), 10⁷ (T2), 10⁸ (T3), 10⁹ (T4), 10¹⁰ (T5) cfu g^-1 of L. plantarum and control group (T6) without diet containing probiotics were fed for 60 days. Results showed that final weight, final length, growth rate, percent weight were gained in treatment 2 the highest and 5 the lowest level in treated. Also, FCR lowest rates in treatment 2 and treatment 5 were accounted for most (p<0.05). The highest total count of lactic acid bacteria were obtained in the intestine of T4(p<0.05). Maximum carcass protein was observed in T4, and low fat content is related to the control treatment (p<0.05). According to the results obtained from the use of Lactobacillus plantarum could be considered as a positive factor for the improvement of the intestinal bacterial flora, growth performance and carcass composition could be used.  

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  The aim of this study was to demonstrate the inhibitory capacity of two strains of  gram positive bacilli, isolated from intestinal content of Persian sturgeon, against Listeria monocytogenes growth. Two strains Lactobacillus casei AP 8 and Lactobacillus plantarum A P 12 , were screened for their antilisterial activity against.  L. monocytogenes, using a disk diffusion agar test. However, L. casei AP 8 always had the highest inhibitory effect. The spoiling potential and antilisterial capacity of bacterial strains was tested in sterile cold smoked roach (CSR) blocks inoculated with 10⁴ CFU g^− 1 of lactic bacteria and 10² CFU g^-1 of Listeria monocytogenes and then stored for 10 days at 4 °C followed by 30 days at 20 °C. L. casei AP8 grew a little faster L. plantarum A P 12 and none of them showed any adverse effect on quality of the product ( i.e. no total volatile basic nitrogen (TVBN) production and no acidification. Lactobacillus casei AP8 was the most efficient strain, maintaining the level of L. monocytogenes at <50 CFU/ g  during  40 dayss of storage at 4 and 20°C. In conclusion, biopreservation of cold smoked roach using bacterial cultures such as L. casei AP8  is a promising way to inhibit the growth of pathogenic bacteria such as L. monocytogenes with low effect on the product quality.

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Surfactin is one of the most efficient biosurfactants excreted by Bacillus subtilis which displays the highest potential as induced systemic resistance elicitor among all metabolites produced by B. subtilis. Environmental factors have considerable effect on surfactin production. In this study surfactin production of two Bacillus subtilis strains were analyzed using high performance liquid chromatography (HPLC). C14 and C15 surfactins were detected in the ethanol extract from acid-precipitated supernatant. HPLC analyses of different media including Nutrient Broth (NB) medium, NB plus 40g/l glucose, NB plus 10% soil extract and NB plus 10% plant extract medium, clearly showed that these bacteria produced different amounts of surfactins C14 and C15 in these media. Surfactin production in NB/plant medium was relatively the highest in quantity. Microelements analysis of media containing plant and soil extract with atomic absorption spectrometry showed high amounts of Fe, Mn and Zn in medium containing plant extract compared with that of soil extract. Since these elements play an important role in surfactin production, high amounts of Fe, Mn and Zn in NB/plant extract medium compared to the NB/soil extract medium could be the possible reason for relatively higher amounts of surfactins C14 and C15 produced in NB/plant medium. So adding these important elements to soil may boost biocontrol effect of B. subtilis against plant pathogens.

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A total of 78 rhizobacterial strains were isolated from 48 rhizospheric soil and root samples, collected from safflower Carthamus tinctorius L. fields located in different regions of Iran. The chitinolytic activity was measured in the presence of colloidal chitin as the sole carbon source. Eleven isolates were identified as chitinolytic bacteria, based on the formation of a clearly visible zone on the growth media. Four isolates including EM9, ES41, ES7 and ER13 exhibited the highest chitin degradation activity based on a clear zone diameter of more than 10 mm. According to a ribotyping analysis, EM9, ES41, and ES7 isolates were identified as Bacillus cereus and ER13 was found to be Pantoea agglomerans. In a dual-culture assay, morphogenic changes such as severely collapsed hyphae, decreased hyphal diameter with condensation and granulation of cytoplasm and highly rolled with formation of big clamydoconida in anomalous sporodochia -like structures were also observed using light microscope. Under greenhouse conditions, the application of selected chitinolytic isolates, i.e., EM9, ES41, ES7 and ER13, on safflower seeds significantly reduced seedling damping-off caused by Fusarium solani. In addition, the results revealed that root and shoot dry weight in infected plants that were treated with EM9 isolate suspension, increased by 14 and 22%, respectively.

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Aims: Burn infections are one of the most common causes of mortality in the world. On the other hand, microbial resistance to antibiotics has caused concern in the medical community. Therefore, controlling burn infections is very important, and using alternative therapies instead of antibiotics could be a good solution to this problem.
Materials & Methods: Lactobacillus plantarum 299v strains were used in the experiments. Fifty male Wistar rats were prepared, and burn was induced in animals. The burn wounds were inoculated with clinical strains of MDR Pseudomonas aeruginosa in all animals and then treated daily with an eucerin ointment containing different compositions, including NaCl, imipenem, probiotic cell pellet, probiotic supernatant, and probiotic cell pellet + probiotic supernatant. The wound healing process was evaluated in animals after 7 days of treatment. Comparisons between different groups were performed using One-way ANOVA and Turkey’s post hoc tests.
Findings: After 7 days of treatment, the mean wound size in the probiotic cell pellet group was significantly lower than in the control and imipenem groups. Also, the mean wound size in the probiotic supernatant group was significantly lower than in the imipenem group. Histological parameters related to skin repair in the probiotic cell pellet group was better than in the control and antibiotic groups. Also, inflammation in the probiotic cell pellet group was less than in the control and imipenem groups.
Conclusion: The macroscopic results of this study supported the microscopic results and showed that the mean size of the burn wounds in the probiotic cell pellet group was less than in the control and imipenem groups after 7 days of treatment.

 

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In the present work, survival of two most important commercial strains of probiotic bacteria, i.e.,  Lactobacillus acidophilus (La5) and Bifidobacterium lactis (Bb12), during production and cold storage of Iranian Doogh, containing Ziziphora extract was studied. The bacteria inoculated into three types of Doohg, a plain sample as the control one, and two samples containing 1% and 2% Ziziphora extract. Survival of probiotic bacteria, pH, acidity and organoleptic characteristics of bio- Doogh were examined during a nine-week cold storage time (4°C). Our results revealed that the population of viable B. lactis reduced by 2.5 log cfu/ml, while L. acidophilus count reduced to zero after eight weeks at 4°C. The pH and acidity of bio-Doogh were not changed significantly (p>0.05) during cold storage period. Also, the organoleptic characteristics of the studied samples changed significantly (p<0.05). Bio-Doogh with Ziziphora extract had higher flavor scores than bio-Doogh without Ziziphora extract.    

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The effects of total carbohydrate, total protein, carbohydrate/protein ratio, amino acid contents, initial pH and aeration on biomass yield product of Bacillus thuringiensis subsp. kurstaki was investigated. The bacteria were cultured in economical media comprising agricultural products and by-products including fishmeal, cottonseed meal, defatted soybean meal, cornsteep liquor, yeast extract, scotafeme, sorghum and peptone as protein sources and glucose and beet molasses as carbohydrate sources. The results indicated the presence of a direct correlation between yield biomass and carbohydrate/protein (C/N) ratio as well as the glutamic acid content of the media. The highest biomass was produced in the media providing 0.4 – 0.5 C/N ratio and 13.9% glutamic acid. A pH range of 7.0 to 8.0 was needed for high yield production. The optimal ratio for culture volume to flask size and shaking speed were 1/5 and 250 rpm, respectively. The biochemical factors described can be considered as the minimal criteria to evaluate culture media for biomass production from Bacillus thuringiensis subsp. kurstaki.

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Endoglucanase Cel9A from Alicyclobacillus acidocaldarius (AaCel9A), a thermophile enzyme, randomly breaks β1-4 glycosidic bond between glucose units in cellulose polymer and produces oligosaccharides with reducing end. In this study, first of all, E.coli BL21 cells were transformed by pDEST17 carrying AaCel9A enzyme gene for expression of the recombinant enzyme. After expression, the recombinant enzyme was purified by Ni-NTA affinity chromatography column and the purity of the recombinant protein was analyzed by SDS-PAGE. Due to impact of the calcium, pH and temperature on AaCel9A activity, the effects of these parameters were investigated on AaCel9A activity to optimize activity condition by using Response surface methodology. The SDS-PAGE result showed that AaCel9A, with molecular weight of 59 kDa, was expressed and purified. Response surface methodology data reveal that the effect of pH on the activity of the enzyme is higher than temperature and the calcium effect is less than temperature. Results showed that the optimum condition of AaCel9A activity reaches at pH 6.35 and 64.5 ˚C as well as 4.92 mM of calcium. Finally, the high correlation between experimental and predicted date indicated that the proposed model for optimizing the enzyme activity has a high accuracy.

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Production and characterization of bacterial thermophilic avicelase Fatemeh Azadian, Arastoo Badoei-dalfard*, Abdolhamid Namaki-Shoushtari, Mehdi Hassanshahian Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran Nowadays, developing processes for effectively converting agricultural wastes for production of high value chemicals has gained considerable interest. Avicelases are important industrial enzymes for the most bioconversion processes. In this study, samples were picked up and inoculated in AVI broth for 7 days at 50 ºC. The bacterial strains with the clear halo (represent extracellular avicelase) have been purified. AV8 isolate which showed the highest clear halo was selected for further studies. This strain was identified as Bacillus genus based on biochemical tests and 16S rRNA analysis. Avicelase production was considered under varying environmental parameters. The best carbon and nitrogen sources for maximum avicelase production were 0.5% sucrose and 0.25% yeast extract, respectively. Avicelase from this strain has been partially purified using ammonium sulphate fractionation followed by dialysis and ion exchange Q-Sepharose chromatography. Results showed that enzyme was active and stable between 30-70 ºC and itʼs maximum temperature activity was observed in 70 ºC. The optimum avicelase activity and stability was observed at pH 6.0. These are characteristics indicating that this enzyme could be an acidophilic and thermophilic avicelase. Furthermore, the avicelase activity improved by methanol (138 %) and chloroform (107 %). These results indicated that AV8 avicelase has potential applications in various industries.

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Ethanol as a renewable biofule is an appropriate and viable alternative to the challenging fossil fuels. Bacillus subtilis, a gram positive bacterium, seems to be a promising choice since it has many useful features. For example B.subtilis ferments broad range of sugars derived from lignocellulosic hydrolysis. Transformation of this cellulytic bacterium to an ethanologenic one was accomplished via metabolic engineering techniques and Ethanol production operon of Z.mobilis was introduced to the B.subtilis. SR1 and SR21 strains expressed plasmid-borne ethanologenic genes of Z.mobilis but the genes had been integrated into the SR22 genomic DNA. Also lactate dehydrogenase gene had been knocked-out in SR21 and SR22 strains. Defect of cell growth in SR21 and SR22, suggests that NAD+ oxidation by lactate dehydrogenase is important for anaerobic growth. Considering the impact of Fe2+ ion on alcohol dehydrogenase II activity, in further experiments Fe2+ was added to the culture media and improvement in growth rates was seen. Final yield of ethanol production of SR1, SR21, and SR22 strains were 53.8%, 86.7%, and 83.9% respectively.