Publication Date: 2024
Coatings (20796412)14(12)
The increasidng demand for implants due to the aging populations highlights the necessity for applying highly functional coatings on the surface of implants. This study investigates the implications of applying a chitosan/polylysine composite coating on anodized titanium surfaces, aiming for improved biocompatibility, bioactivity, and anti-bacterial properties. Titanium substrates were anodized at 40 volts for a duration of two hours, followed by dip coating with the chitosan/polylysine composite. Fourier-transform infrared (FTIR) analysis was employed to characterize the polymer structure, while field emission scanning electron microscopy (FESEM) and energy-dispersive X-ray spectroscopy (EDS) techniques were utilized to evaluate nanotube morphology and the coating structure. Results showed that samples containing 1.5% polylysine exhibited noticeable anti-bacterial properties and cell viability above fifty percent. Subsequent immersion in simulated body fluid (SBF) for a duration of two weeks revealed the formation of apatite crystals on the coated samples, indicating that the samples are bioactive. Furthermore, polylysine contributed to enhanced resistance against degradation in phosphate-buffered saline (PBS) solution. Overall, the chitosan/polylysine composite coating exhibited promising mechanical and biomedical characteristics, suggesting its potential for applications in orthopedic implants. © 2024 by the authors.
Publication Date: 2024
Surfaces and Interfaces (24680230)45
The aim of this study was to develop multi-functional bioactive glass-based coatings with anti-cancer properties for biomedical applications. These coatings were specifically designed for AISI 316 stainless steel substrates. The surface was initially coated with a first layer comprising a composite of iron oxide nanoparticles and 45S5 bioactive glass, which was deposited on the surface through electrophoretic deposition. Subsequently, a second layer, consisting of Pectin and Pectic oligosaccharide polymers, was applied onto the base layer using the immersion technique. Various analytical techniques, including scanning electron microscope, X-ray diffraction, attenuated total reflectance fourier-transform infrared, energy-dispersive X-ray spectroscopy, vibrating sample magnetometry, specific loss power, and inductively coupled plasma, were employed to analyze the synthesized samples. In addition, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) tests, MCF-7 cell culture studies, and corrosion evaluations were also conducted. The vibrating sample magnetometry analyses revealed that the samples exhibited the highest saturation magnetization when synthesized using a Fe3O4 solution with a concentration of 0.5 g L−1. Specific Loss Power analysis indicated that a temperature of 42 °C could be attained within 175 s for the sample prepared under these conditions. The results demonstrated that all the coatings were found to possess hydrophilic surfaces. Examination of MCF-7 cell culture revealed that coatings with a pectic oligosaccharide top layer were found to exhibit remarkable cytotoxicity, leading to the destruction of up to 98.8 % of cancer cells within a two-hour exposure period at 48 °C. © 2024 Elsevier B.V.
Publication Date: 2023
Clinical Nutrition Open Science (26672685)47pp. 6-43
The novel coronavirus infection (COVID-19) conveys a serious global threat to health and economy. A common predisposing factor for development to serious progressive disease is presence of a low-grade inflammation, e.g., as seen in diabetes, metabolic syndrome, and heart failure. Micronutrient deficiencies may also contribute to the development of this state. Therefore, the aim of the present study is to explore the role of the nutrition to relieve progression of COVID-19. According PRISMA protocol, we conducted an online databases search including Scopus, PubMed, Google Scholar and web of science for published literatures in the era of COVID-19 Outbreak regarding to the status of nutrition and COVID-19 until December 2021. There were available studies (80 studies) providing direct evidence regarding the associations between the status of nutrition and COVID-19 infection. Adequate nutritional supply is essential for resistance against other viral infections and also for improvement of immune function and reduction of inflammation. Hence, it is suggested that nutritional intervention which secures an adequate status might protect against the novel coronavirus SARS-CoV-2 (Severe Acute Respiratory Syndrome - coronavirus-2) and mitigate its course. We also recommend initiation of adequate nutritional supplementation in high-risk areas and/or soon after the time of suspected infection with SARS-CoV-2. Subjects in high-risk groups should have high priority for applying this nutritive adjuvant therapy that should be started prior to administration of specific and supportive medical measures. © 2022
Publication Date: 2023
Microbiology (United Kingdom) (13500872)169(7)
Over the past decades, antibiotic resistance has become a major clinical problem, and searching for new therapeutic strategies seems to be necessary. Using novel natural compounds, antimicrobial peptides, and bacteriophages is the most promising solution. In this study, various cationic metabolite-producer bacteria were isolated from different soil samples. Two isolates were identified as Stenotrophomonas maltophilia HS4 (accession number: MW791428) and Paenibacillus polymyxa HS5 (acces-sion number: MW791430) based on biochemical characteristics and phylogenetic analysis using 16S rRNA gene sequences. The cationic compound in the fermentation broth was precipitated and purified with sodium tetraphenylborate salt. The purified cationic peptide was confirmed to be epsilon-poly-l-lysine by structural and molecular analysis using High-Performance Liquid Chromatography, Sodium dodecyl-sulfate-polyacrylamide gel electrophoresis, and Fourier-transform infrared spectroscopy. The antibacterial activity of epsilon-poly-l-lysine was evaluated against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Enterococcus faecalis ATCC 29212, Serratia marcescens ATCC 13880, and Klebsiella pneumoniae ATCC 13883 by microdilution method. Furthermore, the antibacterial effects of purified epsilon-poly-l-lysine in combination with two long non-contractile tail bacteriophages against vancomycin-resistant Enterococcus faecalis and colistin-resistant Klebsiella pneumoniae were investigated. The results indicated great antibacterial activity of epsilon-poly-l-lysine which was produced by two novel bacteria. The epsilon-poly-l-lysine as a potent cationic antimicrobial peptide is demonstrated to possess great antimicrobial activity against pathogenic and also antibiotic-resistant bacteria. © 2023 The Authors.
Publication Date: 2023
Materials Letters (0167577X)352
This paper investigates the antibacterial properties of anthracite/CuO composites. CuO nanoparticles were synthesized and deposited on anthracite particles by chemical bath deposition method. Structural and phase characteristics of synthesized nanoparticles were investigated by X-Ray diffraction (XRD), scanning electron microscope (SEM), and energy-dispersive X-ray spectroscopy (EDS). Results showed that the proposed synthesis method ended up in a homogeneous distribution of CuO nanoparticles, anchored over anthracite particles. Synthesized anthracite/CuO composite showed promising antibacterial properties against Escherichia coli (ATCC 8739) and Staphylococcus aureus (ATCC 6538). © 2023 Elsevier B.V.
Publication Date: 2023
Scientific Reports (20452322)13(1)
Recently Bacillus spp. has gained much attention as potential probiotics due to the production of resistant cells. So, this research is purposeful for evaluation of probiotic characteristics of Bacillus isolates from camel milk as a suitable source for growth and isolation of microorganisms that can be candidate to be used as probiotic. First, forty-eight colonies were screened by using morphological and biochemical analysis. Among the isolates, two of them were recognized as Bacillussubtilis CM1 and CM2 by partial 16SrRNA sequencing that, probiotic potentials of them were evaluated. Both of them, in the preliminary safety screening, were found negative for hemolysis and lecithinase activity. Also, in vitro characteristics such as acid, bile salts and artificial gastric juice resistant, cell surface hydrophobicity, auto-aggregation, antioxidant characteristics, and adherent capability to HT-29 cells were determined for them approximately in the range of other probiotic strains. Two strains were susceptible to various antibiotics and enterotoxigenic activities were not detected by PCR which means isolated Bacillus strains could be classified as safe. Altogether, results demonstrate that Bacillus CM1 and CM2 strains could have the potential of consideration as probiotics, however more extensive in vitro/vivo studies are needed. © 2023, The Author(s).
Publication Date: 2023
Materials (19961944)16(13)
Modification of paint with nanoparticles (NPs) provides self-cleaning, water/dirt-repellent, and other properties. Therefore, the aim of the present study was to biosynthesize silver (Ag) and copper oxide (CuO) NPs and to prepare NP-modified paint. To this end, AgNPs and CuONPs were biosynthesized using Bacillus atrophaeus spores and commercial and crude dipicolinic acid (DPA) extracted from the spore of this bacterium. The synthesized NPs were characterized using electron microscopy, Fourier-transform infrared (FTIR), X-ray diffraction analysis (XRD), and energy-dispersive X-ray spectroscopy (EDS) methods. A minimum inhibitory concentration (MIC) assay of NPs against Escherichia coli ATCC8739 and Staphylococcus aureus ATCC6538 was carried out. The antibacterial effects of prepared NP–paint complexes were assessed using an optical density (OD) comparison before and after adding metal sheets coated with NP–paint complexes into the nutrient broth medium. Four different types of NPs were synthesized in this research: AgNPs synthesized by spore (A), AgNPs synthesized by commercial DPA (B), AgNPs synthesized by crude DPA (C), and CuONPs synthesized by spore (D). SEM analysis confirmed the spherical shape of NPs. According to the results, NPs A, B, and D showed higher antibacterial activity against S. aureus compared to E. coli. Furthermore, the analysis of the antibacterial effects of NP–paint complexes suggested that paint–NPs A, B, and C displayed higher activity on E. coli compared to S. aureus. Moreover, the antibacterial effect of paint–NP D was significantly lower than other NPs. According to this robust antibacterial effect on pathogenic bacteria, it seems that these NP–paint complexes could be useful in public places such as hospitals, airports, dormitories, schools, and office buildings, where the rate of transmission of infection is high. © 2023 by the authors.
Publication Date: 2023
Iranian Journal Of Veterinary Research, Shiraz University (17281997)24(1)pp. 65-73
Background: Species of the Bacillus genus have a long history of use in biotechnology. Some Bacillus strains have recently been identified for food applications and industrial as safe bacteria, which mostly have been recognized as probiotic strains. Aims: The primary purpose of the current study was to evaluate the probiotic characteristics of Bacillus subtilis strains isolated and identified from the goat milk samples. Methods: After sampling from 40 goat milk and cultivation, suspected colonies were subjected to biochemical and molecular identification. Then, the confirmed isolate was assessed for in vitro probiotic tests, including hemolysis and lecithinase properties, bile salt, acid, and artificial gastric juice resistance, antioxidant activity, antibiotics susceptibility, enterotoxin genes detection, and attachment capacity to the HT-29 cells. Results: Among 11 suspected isolates evaluated, only one isolate was identified as B. subtilis. In vitro tests for this strain showed similar results to other probiotic strains. The B. subtilis strain was susceptible to various antibiotics. The enterotoxin genes were not detected based on PCR assay. Concerning its probiotic characteristics assessment, especially tolerance to bile salts and acidic conditions, the Bacillus strain could have the potential to consider as a probiotic. Conclusion: Goat milk can be recommended as a source of Bacillus isolates. Also, the isolated strain showed high adaptability to the gastrointestinal environment, relatively equal percentages of adhesion properties, and some safety aspects, having the potential to be considered as an appropriate probiotic. © 2023 Shiraz University. All rights reserved.
Publication Date: 2022
Biotechnology and Bioprocess Engineering (12268372)27(4)pp. 586-595
Epsilon-Poly-L-lysine (ε-PL) is a non-toxic cationic homopolypeptide with unique bioactive properties. In this study, two novel polylysine producers, Paenibacillus polymyxa HS6 and Stenotrophomonas maltophilia YS8 were isolated from different soil samples and identified by 16S rDNA sequence analysis. To obtain pure cationic polypeptide from these bacteria, chemical precipitation with sodium tetraphenylborate was used. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, High-performance liquid chromatography, and Fourier-transform infrared spectroscopy confirmed the purified cationic compound to be polylysine. The antioxidant activity of ε-PL was determined by the 2,2-diphenyl-1-picryl-hydrazyl-hydrate method. Moreover, the anticancer and cytotoxic effects were evaluated against MCF-7, HT-29, and L929 cells by thiazolyl blue tetrazolium bromide and flow cytometry. The maximum yields of 1.8 and 0.39 g/L polylysine were obtained by P. polymyxa HS6 and S. maltophilia YS8, respectively. The results showed that radical scavenging activity of polylysine at a minimum concentration (0.18 mg/mL) was at least 7% which increased to 90% at 8 mg/mL. Furthermore, the highest anticancer activity was observed against the MCF-7 cells (99.5%) at 0.5 mg/mL concentration, while almost no toxicity was observed towards L929 cells. Based on the results obtained in this study, it is suggested that the polylysine produced by P. polymyxa HS6 and S. maltophilia YS8 is a potential bioactive compound with significant anticancer and antioxidant properties. © 2022, The Korean Society for Biotechnology and Bioengineering and Springer.
Publication Date: 2022
Scientific Reports (20452322)12(1)
Pectin is one of the main structural components in fruits and an indigestible fiber made of d-galacturonic acid units with α (1-4) linkage. This study investigates the microbial degradation of pectin in apple waste and the production of bioactive compounds. Firstly, pectin-degrading bacteria were isolated and identified, then pectinolytic activity was assessed by DNS. The products were evaluated by TLC and LC–MS–ESI. The antioxidative effects were investigated using DPPH and anti-cancer effects and cytotoxicity were analyzed by MTT and flow cytometry. In this study two new bacterial isolates, Alcaligenes faecalis AGS3 and Paenibacillus polymyxa S4 with the pectinolytic enzyme were introduced. Structure analysis showed that the products of enzymatic degradation include unsaturated mono, di, tri, and penta galacturonic acids with 74% and 69% RSA at 40 mg/mL for A. faecalis and P. polymyxa S4, respectively. The results of anti-tumor properties on MCF-7 cells by MTT assay, for products of AGS3 and S4 at 40 mg/mL after 48 h, showed 7% and 9% survival, respectively. In the flow cytometric assessment, the compounds of AGS3 at 40 mg/mL were 100% lethal in 48 h and regarding S4 isolate caused 98% death. Cytotoxicity evaluation on L-929 cells showed no significant toxicity on living cells. © 2022, The Author(s).
Publication Date: 2022
Environmental Research (00139351)207
This paper investigates the synthesis, antibacterial, and photocatalytic properties of silver ion-exchanged natural zeolite/TiO2 photocatalyst nanocomposite. Zeolite is known to have a porous surface structure, making it an ideal substrate and framework in different nanocomposites. Moreover, natural zeolite has a superior thermal and chemical stability, with hardly any reactivity with chemicals. Finding an effective and low-cost method to remove both antibiotics and bacteria from water resources has become a vital global issue due to the worldwide excessive use of chemicals and antibiotics. This research aims to propose a facile method to synthesize Ag-ion-exchanged zeolite/TiO2 catalyst for anti-bacterial purposes and photocatalytic removal of atibiotics from wastewaters. TiO2 particles were deposited on the surface of natural zeolite. Ag ion exchanging was performed via a liquid ion-exchange method using 0.1 M AgNO3 solution. X-ray diffractometry (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and Fourier-transform infrared spectroscopy (FTIR) were used to evaluate the structure of synthesized powders. Antibacterial activities of samples were assessed, using Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 by disc diffusion method. It was shown that Ag-containing nanocomposite samples have an improved antibacterial performance in both cases. Results showed that the synthesized catalyst has promising potentials in wastewater treatment. © 2021 Elsevier Inc.
Publication Date: 2021
Scientific Reports (20452322)11(1)
Pectin, a diverse carbohydrate polymer in plants consists of a core of α-1,4-linked D-galacturonic acid units, includes a vast portion of fruit and agricultural wastes. Using the wastes to produce beneficial compounds is a new approach to control the negative environmental impacts of the accumulated wastes. In the present study, we report a pectinase producing bacterium Streptomyces hydrogenans YAM1 and evaluate antioxidative and anticancer effects of the oligosaccharides obtained from pectin degradation. The production of oligosaccharides due to pectinase activity was detected by thin layer chromatography (TLC) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Our results revealed that S. hydrogenans YAM1 can degrade pectin to unsaturated pectic oligo-galacturonic acids (POS) with approximately 93% radical scavenging activity in 20 mg/mL which it is more than 50% of the same concentration of pectin. Flow cytometric analysis revealed that MCF-7 cells viability decreased more than 32 and 92% following treatment with 6 and 20 mg/mL POS after 24 h, respectively. It is suggested that pectin degradation by S. hydrogenans YAM1 is not only a new approach to produce highly active compounds from fruit wastes, but also is an effective method to remove fibrous pollutants from different environments. © 2021, The Author(s).
Publication Date: 2021
Ceramics International (02728842)47(23)pp. 33875-33885
This paper investigates the synthesis and characterization of photoactive TiO2/CuO nanocomposites for the simultaneous antibacterial applications and photocatalytic removal of Amoxicilline from wastewaters. Effective removal of biological and organic contaminants from water resources has become a global challenge due to contaminants' complexity and extensive use. TiO2/CuO heterojunctions with different CuO loadings were synthesized using a straightforward precipitation method. Electron microscopy, energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), Fourier transformed infrared (FTIR), ultraviolet–visible (UV-VIS) analyses were used to characterize synthesized samples. Antibacterial activities of samples were investigated against Staphylococcus aureus. Also, the applicability of synthesized powders in the photocatalytic removal of Amoxicilline from wastewater was methodically investigated. Results showed that the photocatalytic activity of synthesized TiO2/CuO nanocomposites is highly dependent on the CuO loadings of samples. CuO loading can also increase the light absorption within the visible light region, making TiO2/CuO samples applicable in the visible light region. Results also showed that CuO loading greatly enhances the antibacterial characteristics of samples. © 2021
Publication Date: 2020
Journal of Alloys and Compounds (09258388)821
Copper oxide particles have found applications as strong photocatalytic antibacterial agents in recent years. This paper investigates the facile chemical bath deposition synthesis and antibacterial properties of nanocrystalline CuO, CuO-GO, and CuO-GO-Ag nanocomposites. CuO samples were synthesized through a chemical bath deposition method and the introduction of GO nanosheets and silver nanoparticles into the precursor solutions was used to obtain CuO-GO and CuO-GO-Ag nanocomposites. Scanning electron microscope, energy dispersive spectroscopy, elemental mapping, and Fourier-transform infrared spectroscopy were used to characterize synthesized specimens. Antibacterial properties of the nanocomposites were examined against Gram-positive Staphylococcus aureus ATCC 25923 and Gram-negative Escherichia coli ATCC 25922 by the disc diffusion method and minimum inhibitory concentration (MIC) measurements. CuO particles with monoclinic crystallographic structure and crystallite size of 10 nm were synthesized. Synthesized particles were spherical with a narrow size distribution, between 5 and 10 μm. Minor addition of GO to the precursor solution resulted in less agglomeration of nanocomposite powders. CuO-GO-Ag nanocomposite showed noticeable enhanced antibacterial activities against the tested bacteria under visible light, resulting in MIC of 2.6 ± 0.5 mg/ml against Gram-positive and Gram-negative bacteria and also higher inhabitation zone compared to CuO and CuO-GO nanocomposites. © 2019 Elsevier B.V.
Publication Date: 2020
Current Microbiology (03438651)77(1)pp. 40-48
Some patients with candidiasis seek alternatives drug to treat vaginal yeast infection like herbal preparations and probiotics. However, the effectiveness of such treatments has not received much study. In this research, the unique chitinotrophic Bacillus was isolated on shrimp shell from salt lakes and identified as Bacillus altitudinis by 16SRNA sequencing. This strain produced a novel chitin-oligosaccharide material and thermostable chitinase (5.1 units/ml) during 4 days incubation on shrimp shell medium; nevertheless, its growth on nutrient agar was negative. The zymogram showed less than 50 kD protein responsible for chitinase activities. The LC/MS detection of concentrate fermented products showed the production of oligosaccharide during chitin fermentation. As results of shrimp shell degradation, 65.6 mg/l protein, 73.4 mg/l N-acetyl glucose amine, and oligosaccharide were produced. Synergism activities of chitooligosaccharide and chitinase from this strain against fungi and pathogen candida (staining with methylene blue showed that almost 50% of 106 cells were died during 6 h) are promising for new anti-fungal drug with no side effect. © 2019, Springer Science+Business Media, LLC, part of Springer Nature.
Publication Date: 2019
Biotechnology and Bioprocess Engineering (12268372)24(3)pp. 507-512
Orobol (5,7,3′,4′-tetrahydroxyisoflavone) is a highly hydroxylated isoflavone, which is rarely found in natural environment. In this study, orobol was produced due to bioconversion of a soybean frequented isoflavone, genistein (4′,5,7-Trihydroxyisoflavone) by an active, stable, reusable genetically immobilized enzyme, the recombinant Bacillus subtilis spore displayed tyrosinase. Thin layer chromatography and high performance liquid chromatography were used to monitor the reaction. The results revealed that 1 mM orobol was produced from 3′-hydroxylation of 1 mM genistein by spore displayed tyrosinase at 37°C during 90 min incubation. To study on anti-proliferative effects of orobol, MCF-7 breast cancer cell viability was determined by MTT method and flow cytometric analysis. The comparison between reduction in cell viabilities in 50 to 500 µM genistein and orobol treated cells revealed that orobol has more remarkable anticancer effects than genistein. Flow cytometric analysis showed more than 87% cytotoxicity in 500 µM orobol treated cells by flow cytometric analysis. To the best of our knowledge this is the first report of the orobol demonstrated potent anticancer activity against MCF-7 breast cancer cell. It is suggested that enzymatic biotransformation of soybean genistein to orobol will be made a new approach to create highly bioactive products usable in food and pharmaceutical industries. © 2019, The Korean Society for Biotechnology and Bioengineering and Springer.
Publication Date: 2019
Journal of Basic Microbiology (0233111X)59(3)pp. 249-255
Tragacanth, a highly branched carbohydrate polymer isolated from Astragalus, is one of the most commonly used gums in food industry. The primary structure of tragacanth is composed of galacturonic acid monomers connected with α 1–4 links, and it is very similar to the pectin. Tragacanth degradation by microorganisms is significant in two aspects: first, food preservation and microbial growth control due to too much use of tragacanth in the food industry, second, therapeutic and pharmaceutical potential of obtained oligosaccharides. In the present study, we report three new strains of bacteria, Acinetobacter guillouiae strain TD1, Kosakonia sacchari strain TD2, and Bacillus vallismortis strain PD1 with the capability of growing in tragacanth as an only source of carbon and energy. The evolutionary history of the isolated strains was analyzed based on 16S rRNA gene sequences in MEGA7 using the neighbor-joining method. The production of di and tri galacturonic acid due to pectinase activities of the strains were detected by thin layer chromatography (TLC) and liquid chromatography/Mass spectroscopy (LC/MS) analysis. Here is the first report of the ability to grow in tragacanth and pectinase activity monitoring in bacteria. Our results revealed that all of the isolated strains are capable of degrading pectin and tragacanth to oligo-galacturonic acids. The obtained products, which have different structures depending on the tragacanth structures and types of pectinolytic enzymes, would show therapeutic and pharmaceutical potentials. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Publication Date: 2019
Metals (20754701)9(4)
This paper investigates a severe microbiologically influenced failure in the elbows of a buried amine pipeline in a petrochemical plant. Pipelines can experience di_erent corrosion mechanisms, including microbiologically influenced corrosion (MIC). MIC, a form of biodeterioration initiated by microorganisms, can have a devastating impact on the reliability and lifetime of buried installations. This paper provides a systematic investigation of a severe MIC-related failure in a buried amine pipeline and includes a detailed microstructural analysis, corrosion products/biofilm analyses, and monitoring of the presence of causative microorganisms. Conclusions were drawn based on experimental data, obtained from visual observations, optical/electron microscopy, and Energy-dispersive X-ray spectroscopy (EDS)/X-Ray Di_raction (XRD) analyses. Additionally, monitoring the presence of causative microorganisms, especially sulfate-reducing bacteria which play the main role in corrosion, was performed. The results confirmed that the failure, in this case, is attributable to sulfate-reducing bacteria (SRB), which is a long-known key group of microorganisms when it comes to microbial corrosion. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
Publication Date: 2019
Advanced Pharmaceutical Bulletin (22285881)9(2)pp. 331-334
Purpose: Simple and cheap diagnostic kit development is one of the important aims of pharmaceutical developers and companies focused on public health improvement. The Bacillus subtilis spore surface-display technique is a genetic engineering method that is used to develop new-generation diagnostic kits applicable for the early detection of various types of diseases. In this study, we developed a novel simple, rapid, and inexpensive diagnostic paper-based kit to detect tyrosine in urine samples of humans and animals that is applicable for home or laboratory use. Methods: The B. subtilis spore-displayed tyrosinase system developed by genetic engineering methods was used to prepare a paper-based kit to detect tyrosine in urine samples of different groups of patients (i.e., patients with diabetes, diabetes with chronic kidney disease (CKD), and chronic kidney disease) for the detection of tyrosine during the acute disease phase. To confirm the sensitivity and specificity of the kit, tyrosine was also detected in urine samples using conventional liquid chromatography/mass spectroscopy. Results: Different concentrations of tyrosine (0.1–1 mM) were detected in urine samples based on visible changes of color from bright brownish-gray to dark brownish-gray within 1 hour. The kit could screen samples to distinguish the three groups of patients based on formation of a broad spectrum of colors reflecting the concentration of tyrosine. Conclusion: To the best of our knowledge, this is the first diagnostic kit with potential to rapidly diagnose various diseases related to the production of tyrosine in biological samples. This kit is not only widely applicable, including for personal use in the home, but is also appropriate as a part of standard screening tests and health protection programs in countries with limited resources. © 2019 The Author (s).
Publication Date: 2019
Biotechnology and Bioprocess Engineering (12268372)24(2)pp. 375-381
In this paper, we report removing heavy metal using Bacillus subtilis spore surface display system. We used CotE protein as an anchoring motif because of its high abundance and location in the outer coat layer. And we inserted His12 (double histidine 6 tag) at the C-terminal end of anchoring motif. The proper expression of CotE-His12 fusion protein (22.8 kDa) was confirmed by western blot. We confirmed the surface expression of the CotE-His12 fusion protein using flow cytometry. We tried Ni2+ and Cd2+ adsorption with recombinant spore DB104 (pCotE-His12) and DB104 spore. The amount of adsorbed Ni2+ was 18.2 nmol/mg for DB104 spore and 82.4 nmol/mg for DB104 (pCotE-His12) spore. In the case of Cd2+, the adsorbed amount was 32.6 nmol/mg for DB104 spore and 79.1 nmol/mg for DB104 (pCotE-His12) spore. This means that our spore displayed His12 system can be generally applied for the removal of various kind of heavy metals in the field. © 2019, The Korean Society for Biotechnology and Bioengineering and Springer.
Publication Date: 2016
Journal of Basic Microbiology (0233111X)56(12)pp. 1331-1337
Tyrosinases, copper-containing monooxygenases, are widely used enzymes for industrial, medical, and environmental applications. We report the first functional surface display of Bacillus megaterium tyrosinase on Bacillus subtilis spores using CotE as an anchor protein. Flow Cytometry was used to verify surface expression of tyrosinase on the purified spores. Moreover, tyrosinase activity of the displayed enzyme on B. subtilis spores was monitored in the presence of L-tyrosine (substrate) and CuSO4 (inducer). The stability of the spore-displayed tyrosinase was then evaluated after 15 days maintenance of the spores at room temperature, and no significant decrease in the enzyme activity was observed. In addition, the tyrosinase-expressing spores could be repeatedly used with 62% retained enzymatic activity after six times washing with Tris-HCl buffer. This genetically immobilized tyrosinase on the spores would make a new advance in industrial, medical, and environmental applications. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Publication Date: 2014
Applied Biochemistry and Biotechnology (02732289)174(1)pp. 270-282
Seeking for simple, rapid, and environmental-friendly routes to produce metal nanoparticles is quite attractive for various biotechnological applications. Biological synthesis method of silver nanoparticles has been found very promising due to their non-toxicity and simplicity. Here, the spores of Bacillus stratosphericus isolated from soil enriched with 30 % H2O2 were used for the production of silver nanoparticles. Furthermore, the possible mechanism of silver nanoparticle synthesis by the spores was elucidated for the first time. In this regard, dipicolinic acid (DPA) was shown to play a critical role as a nanoparticle-producing agent. UV–Vis absorption spectroscopy, X-ray diffraction technique, energy-dispersive spectroscopy, and transmission electron microscopy were used to characterize the nanoparticles. Unlike vegetative cells of B. stratosphericus, the spores and the purified DPA were capable of producing nanoparticles from silver nitrate (AgNO3). These biogenic nanoparticles, which were highly toxic against different pathogenic bacteria, showed mixed structures including spherical, triangular, cubic, and hexagonal with the approximate size between 2 and 20 nm in diameter. Our results illustrated the role of dipicolinic acid as a main factor for the synthesis of nanoparticles by the bacterial spores. © 2014, Springer Science+Business Media New York.
Publication Date: 2013
World Journal of Microbiology and Biotechnology (09593993)29(12)pp. 2359-2364
The biological synthesis methods have been emerging as a promising new approach for production of nanoparticles due to their simplicity and non-toxicity. In the present study, spores of Bacillus athrophaeus were used to achieve the objective of developing a green synthesis method of silver nanoparticles. Enzyme assay revealed that the spores and their heat inactivated forms (microcapsules) were highly active and their enzymatic contents differed from the vegetative cells. Laccase, glucose oxidase, and alkaline phosphatase activities were detected in the dormant forms, but not in the vegetative cells. Although no nanoparticle was produced by active cells of B. athrophaeus, both spores and microcapsules were efficiently capable of reducing the silver ions (Ag+) to elemental silver (Ag0) leading to the formation of nanoparticles from silver nitrate (AgNO3). The presence of biologically synthesized silver nanoparticles was determined by obtaining broad spectra with maximum absorbance at 400 nm in UV-visible spectroscopy. The X-ray diffraction analysis pattern revealed that the nanoscale particles have crystalline nature with various topologies, as confirmed by transmission electron microscopy (TEM). The TEM micrograph showed the nanocrystal structures with dimensions ranging from 5 to 30 nm. Accordingly, the spore mixture could be employed as a factory for detoxification of heavy metals and subsequent production of nanoparticles. This research introduces an environmental friendly and cost effective biotechnological process for the extracellular synthesis of silver nanoparticles using the bacterial spores. © 2013 Springer Science+Business Media Dordrecht.
Publication Date: 2013
Jundishapur Journal Of Microbiology (20083645)6(3)
Background: Toluene is a cyclic aromatic hydrocarbon which is widely used as an industrial feedstock and as a solvent. It is one of the major parts of pollution in oil-contaminated environments. Objectives: The main aim of this study was to isolate and characterize a bacterium with high potential application in toluene bioremediation. Materials and Methods: To isolate a toluene-degrading bacterium, several seawater and wastewater samples were added to toluene-containing basal salt media (BSM). The isolate was identified by morphological features, biochemical tests, and molecular characterization. Also, physiological characteristics of the isolated strain were determined. Results: The isolate represented the capability of growing on toluene under both aerobic and anaerobic conditions. Moreover, this bacterium could also use different toxic compounds as the sole sources of carbon and energy. Sequence analysis of 16S rDNA showed that the isolated strain was closely related to Uncultured bacterium clone A1-E3_M13R (98%) and was submitted as Bacterium Ex-DG74 in NCBI. Bacterium Ex-DG74 showed a tolerance to organic solvent and saline conditions as it could grow in the medium containing over 15% toluene (v/v) and NaCl (w/v), and degraded 79% and 45% of toluene (1% (v/v)) in aerobic and anaerobic conditions, respectively. Conclusions: In this investigation we succeeded to isolate a novel toluene-degrading bacterium from wastewater. This isolated strain could be considered as a biological material for the toluene bioremediation. © 2013, Ahvaz Jundishapur University of Medical Sciences.
Publication Date: 2012
Jundishapur Journal Of Microbiology (20084161)5(3)pp. 479-485
Background: Toluene which widely exists in petroleum and its related products has gathered much attention due to its adverse effects on health and carcinogenic potential. Since microorganisms are able to utilize petroleum hydrocarbon as carbon and energy sources, they can be used for bioremediation applications. Objectives: The aim of this study was to isolate toluene degrading bacteria from wastewater and seawater. The production of exopolysaccharide, biosurfactant and peroxidase enzymes such as laccase and catalase were investigated to determine the effect of them on toluene degradation. Materials and Methods: To screen and isolate toluene degrading bacteria, contaminated seawater and wastewater samples were added to toluene containing mineral media (MM). The biochemical and molecular characteristics of the isolates were then studied. Results: From seawater, two toluene degrading Bacillus and one Sporosacina species and from wastewater a novel high capable toluene degrading strain, Bacterium Ex-DG74 were isolated and introduced. Bacterium Ex-DG74 showed tolerance to 15 % (v/v) toluene but the marine isolated species could tolerate only 1 % (v/v) toluene. This bacterium also showed the highest catalase and membrane-bound laccase activity. The spore-forming marine bacterium, S. halophila produced large amounts of exopolysaccharide, biosurfactant and extracellular laccase. Conclusions: The results of the present research indicated that EPS, biosurfactant and peroxidase enzymes can have essential roles on toluene tolerance and biodegradation. These native microbial isolates could be considered as a powerful approach for the in situ bioremediation of hydrocarbon-contaminated sea and wastewater. Copyright © 2012 Kowsar Corp.
Publication Date: 2011
Desalination and Water Treatment (19443994)28(1-3)pp. 8-11
For over three decades, methyl-tert-butyl ether (MTBE) has been used as an additive to either increase the octane number in high and medium grade gasoline in substitution of tetraethyl lead or to raise its oxygen content. However, the fate of MTBE in the environment is a great concern. In this work, biomass of novel toluene degradator Exiguobacterium, isolated from toluene enriched seawater, could utilize MTBE by 72% and produced CO2 and formate during degrada tion. Production of ketone and tert-butyl alchol (TBA) has not been seen by gas chromatogra phy (GC). Degradation of MTBE has been seen in aerobic and anaerobic conditions from 4 to 30 °C. It is interesting that the highest degradation occurred during growth in aerobic and cold condition (4 °C). This suggested that in cold areas Exiguobacterium is a very good candidate to degrade MTBE. Since this novel isolate could tolerate high salt, it is useful for removal of MTBE from seawater. © 2011 Taylor & Francis Group, LLC.
