Pub Date : 2020-12-01DOI: 10.30491/JABR.2020.230789.1225
A. Abd-ElKhalek, D. Seoudi, O. Ibrahim, N. Abd-Rabou, E. M. El-Azeem
Introduction: Proteases are hydrolyzing enzymes and are considered to be one of the most important groups of enzymes for industry and are used in leather, pharmaceutical, and food industry along with detergents, and bioremediation processes. The main objectives of this study were (i) to extract, partially purify, and characterize the protease enzyme from Moringa (Moringa oleifera) leaves; and (ii) to investigate the effect of such an enzyme against some pathogenic bacteria.Materials and Methods: This enzyme was extracted in a 0.1 M phosphate buffer pH 7. It was left for 24 hours in a refrigerator and was then filtered using filter paper Whatman no. 41. The aqueous filtrate was used to estimate the proteolytic activity.Results: Purification by ammonium sulfate gave the best results at 50%-70% concentration which had the highest specific activity, highest purification fold with the percentage yield of 45.3%. Gel filtration by Sephadex G-100 gave the best specific activity and the best purification fold with the yield from fraction of 34%-43%. The protease enzyme has optimum pH 7 and temperature 50 oC. The enzyme was thermally stable at 40-70 oC for 20-30 minutes. Some metal ions were activator on the enzyme-like Mn2+ (highest), Ba2+, Ca2+, and Na+. The efficacy of protease enzyme was improved when integration with antibiotic against certain bacterial including Bacillus cereus (S3), Staphylococcus aureus, Salmonella typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes. Also, E. coli O157:H7, L. monocytogenes, and Yersinia enterocolitica did not show any growth at pH 10.Conclusions: To conclude, it can be stated that protease enzyme can be considered as a promising agent, cheap, and safe source which is suitable for using in various industries.
{"title":"Extraction, Partial Purification, Characteristics, and Antimicrobial Activity of Plant Protease From Moringa Oleifera Leaves","authors":"A. Abd-ElKhalek, D. Seoudi, O. Ibrahim, N. Abd-Rabou, E. M. El-Azeem","doi":"10.30491/JABR.2020.230789.1225","DOIUrl":"https://doi.org/10.30491/JABR.2020.230789.1225","url":null,"abstract":"Introduction: Proteases are hydrolyzing enzymes and are considered to be one of the most important groups of enzymes for industry and are used in leather, pharmaceutical, and food industry along with detergents, and bioremediation processes. The main objectives of this study were (i) to extract, partially purify, and characterize the protease enzyme from Moringa (Moringa oleifera) leaves; and (ii) to investigate the effect of such an enzyme against some pathogenic bacteria.Materials and Methods: This enzyme was extracted in a 0.1 M phosphate buffer pH 7. It was left for 24 hours in a refrigerator and was then filtered using filter paper Whatman no. 41. The aqueous filtrate was used to estimate the proteolytic activity.Results: Purification by ammonium sulfate gave the best results at 50%-70% concentration which had the highest specific activity, highest purification fold with the percentage yield of 45.3%. Gel filtration by Sephadex G-100 gave the best specific activity and the best purification fold with the yield from fraction of 34%-43%. The protease enzyme has optimum pH 7 and temperature 50 oC. The enzyme was thermally stable at 40-70 oC for 20-30 minutes. Some metal ions were activator on the enzyme-like Mn2+ (highest), Ba2+, Ca2+, and Na+. The efficacy of protease enzyme was improved when integration with antibiotic against certain bacterial including Bacillus cereus (S3), Staphylococcus aureus, Salmonella typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes. Also, E. coli O157:H7, L. monocytogenes, and Yersinia enterocolitica did not show any growth at pH 10.Conclusions: To conclude, it can be stated that protease enzyme can be considered as a promising agent, cheap, and safe source which is suitable for using in various industries.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"243-250"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49277487","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-12-01DOI: 10.30491/JABR.2021.122254
Samaneh Rahamooz-Haghighi, K. Bagheri, H. Danafar, A. Sharafi
Introduction: Plantago lanceolata is one the most important species of the Plantago genus and has valuable medicinal secondary metabolites. Materials and Methods: The effect of different factors on germination of P. lanceolata seeds was studied and leaf and root explants of in vitro growth seedling were cultured on Murashige and Skoog (MS) medium supplemented with combination of 6-benzylaminopurine (BAP) or thidiazuron (TDZ) (0, 0.5, 1, 1.5, 2 and 3 mg/L) and auxins: α-naphthaleneacetic acid (NAA) (0, 0.2, 0.5, 1 mg/L), Indole-3-acetic acid (IAA) (0.1, 0.5, 1, 1.5 mg/L) or Indole-3-butyric acid (0.5, 1, 1.5, 2 mg/L) (IBA) at different concentrations. Results: The results showed that cold pre-treatment, daylight and 1/8 MS salt concentration are more suitable for high germination. The best shoot organogenesis rate (95%) in leaf explants was observed in 1:1 mg/L, and 1.5:1 mg/L TDZ: IBA. The highest percentage of shoot organogenesis (100%) was observed in most of the plant growth regulator (PGR) treatments in root explants. About 58.67 and 60 shoot numbers obtained with 2 mg/L TDZ in leaf and 1:1.5 mg/L BAP: IBA in root explants, respectively. Conclusions: It can be suggested that the best shoot organogenesis and proliferation medium is MS basal medium containing cytokinin TDZ and auxin IBA in comparison with other hormone compounds on leaf and root explants. The result behind this fact is high callus induction and regeneration potential of explants in all concentrations.
{"title":"Tissue Culture, In Vitro Organogenesis and Regeneration of Plantago lanceolata","authors":"Samaneh Rahamooz-Haghighi, K. Bagheri, H. Danafar, A. Sharafi","doi":"10.30491/JABR.2021.122254","DOIUrl":"https://doi.org/10.30491/JABR.2021.122254","url":null,"abstract":"Introduction: Plantago lanceolata is one the most important species of the Plantago genus and has valuable medicinal secondary metabolites. Materials and Methods: The effect of different factors on germination of P. lanceolata seeds was studied and leaf and root explants of in vitro growth seedling were cultured on Murashige and Skoog (MS) medium supplemented with combination of 6-benzylaminopurine (BAP) or thidiazuron (TDZ) (0, 0.5, 1, 1.5, 2 and 3 mg/L) and auxins: α-naphthaleneacetic acid (NAA) (0, 0.2, 0.5, 1 mg/L), Indole-3-acetic acid (IAA) (0.1, 0.5, 1, 1.5 mg/L) or Indole-3-butyric acid (0.5, 1, 1.5, 2 mg/L) (IBA) at different concentrations. Results: The results showed that cold pre-treatment, daylight and 1/8 MS salt concentration are more suitable for high germination. The best shoot organogenesis rate (95%) in leaf explants was observed in 1:1 mg/L, and 1.5:1 mg/L TDZ: IBA. The highest percentage of shoot organogenesis (100%) was observed in most of the plant growth regulator (PGR) treatments in root explants. About 58.67 and 60 shoot numbers obtained with 2 mg/L TDZ in leaf and 1:1.5 mg/L BAP: IBA in root explants, respectively. Conclusions: It can be suggested that the best shoot organogenesis and proliferation medium is MS basal medium containing cytokinin TDZ and auxin IBA in comparison with other hormone compounds on leaf and root explants. The result behind this fact is high callus induction and regeneration potential of explants in all concentrations.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"258-265"},"PeriodicalIF":0.0,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45019856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-10-13DOI: 10.30491/JABR.2020.117890
M. Khafaei, M. Sadeghi, Peyman Zargari, Fatemeh Saadatmand, Roghayeh Barouei, A. Mohammadi
Introduction: In historical cases, mass disasters, missing person’s identification, and Archaeogenetic investigations, the bone, teeth, and hair samples are often the best and the only biological material available for DNA typing. In this study, we demonstrated an extremely effective protocol for DNA extraction of hair. Materials and Methods: Obtaining genetic information from hair samples for DNA typing involves the following steps: sterilization, DNA extraction, quality control, PCR, and profiling. After extracting aDNA (Ancient DNA) from the hair, autosomal STR analyses were performed for each sample using the AmpFlSTR® MiniFiler™ and Identifiler™ kits. Result: Qualitative analysis of DNA by electrophoresis showed the excellent quality of the extracted samples so that the electrophoresed bands were clearly visible. Conclusions: The results demonstrated that the FDEH method is cheaper and faster than commercial kits; moreover, this new method enhances DNA recovery from hair. Because of simple protocol and high quality, this method can be utilized in Medical.
{"title":"An Effective Method for Extracting DNA From Human Hair Samples","authors":"M. Khafaei, M. Sadeghi, Peyman Zargari, Fatemeh Saadatmand, Roghayeh Barouei, A. Mohammadi","doi":"10.30491/JABR.2020.117890","DOIUrl":"https://doi.org/10.30491/JABR.2020.117890","url":null,"abstract":"Introduction: In historical cases, mass disasters, missing person’s identification, and Archaeogenetic investigations, the bone, teeth, and hair samples are often the best and the only biological material available for DNA typing. In this study, we demonstrated an extremely effective protocol for DNA extraction of hair. Materials and Methods: Obtaining genetic information from hair samples for DNA typing involves the following steps: sterilization, DNA extraction, quality control, PCR, and profiling. After extracting aDNA (Ancient DNA) from the hair, autosomal STR analyses were performed for each sample using the AmpFlSTR® MiniFiler™ and Identifiler™ kits. Result: Qualitative analysis of DNA by electrophoresis showed the excellent quality of the extracted samples so that the electrophoresed bands were clearly visible. Conclusions: The results demonstrated that the FDEH method is cheaper and faster than commercial kits; moreover, this new method enhances DNA recovery from hair. Because of simple protocol and high quality, this method can be utilized in Medical.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42385073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-10-12DOI: 10.30491/JABR.2020.117863
Sadrollah Mahmoudi, M. Ghane, Mehrdad Faraji, H. Goodarzi, Fahimeh Shahjooie, H. Javadzadeh
Introduction: Recently, the consumption of probiotics to decrease oxidative stress has been recommended. We aimed to assess the effect of probiotics on oxidative stress in various diseases and as well as in different models in a systematic review study. Methods: Articles searched through scientific sources such as PubMed, MEDLINE, Wiley, EMBASE, ISI Web of Knowledge and Scopus by two review authors independently. The keywords that applied to search these articles included: Probiotics, oxidative stress, Stresses, Oxidative and Stress. Then results combined and reviewed regarding the type of study, type of disease/participants and outcomes. Results: We included 19 eligible studies. The results showed that various probiotics supplements could improvement oxidative stress parameters in women pregnant, patients with type 2 diabetics, Diabetic kidney disease type 2, gestational diabetes mellitus, aging process, Inflammatory factors in petrochemical workers, acute necrotizing pancreatitis, and polycystic ovary syndrome. But, no significant effect was reported on the oxidative status in patients with Rheumatoid Arthritis. Conclusions: Probiotics supplements could improvement oxidative stress biomarkers in various diseases. There are still some controversies about these methods and types of probiotics supplementations. So, it is recommended that more studies should be conducted for a better decision.
简介：最近，有人建议食用益生菌来减少氧化应激。在一项系统综述研究中，我们旨在评估益生菌对各种疾病和不同模型中氧化应激的影响。方法：由两位独立的综述作者通过PubMed、MEDLINE、Wiley、EMBASE、ISI Web of Knowledge和Scopus等科学来源检索文章。用于搜索这些文章的关键词包括：益生菌、氧化应激、应激、氧化和应激。然后对研究类型、疾病类型/参与者和结果进行综合和回顾。结果：我们纳入了19项符合条件的研究。结果表明，各种益生菌补充剂可改善孕妇、2型糖尿病患者、2型肾病患者、妊娠期糖尿病患者、衰老过程、石化工人炎症因子、急性坏死性胰腺炎和多囊卵巢综合征患者的氧化应激参数。但是，没有报道对类风湿性关节炎患者的氧化状态有显著影响。结论：益生菌补充剂可以改善各种疾病的氧化应激生物标志物。关于这些益生菌补充剂的方法和类型仍然存在一些争议。因此，建议进行更多的研究，以便做出更好的决定。
{"title":"The Effect of Probiotics Supplements on Oxidative Stress in Various Diseases: A Systematic Review Study","authors":"Sadrollah Mahmoudi, M. Ghane, Mehrdad Faraji, H. Goodarzi, Fahimeh Shahjooie, H. Javadzadeh","doi":"10.30491/JABR.2020.117863","DOIUrl":"https://doi.org/10.30491/JABR.2020.117863","url":null,"abstract":"Introduction: Recently, the consumption of probiotics to decrease oxidative stress has been recommended. We aimed to assess the effect of probiotics on oxidative stress in various diseases and as well as in different models in a systematic review study. Methods: Articles searched through scientific sources such as PubMed, MEDLINE, Wiley, EMBASE, ISI Web of Knowledge and Scopus by two review authors independently. The keywords that applied to search these articles included: Probiotics, oxidative stress, Stresses, Oxidative and Stress. Then results combined and reviewed regarding the type of study, type of disease/participants and outcomes. Results: We included 19 eligible studies. The results showed that various probiotics supplements could improvement oxidative stress parameters in women pregnant, patients with type 2 diabetics, Diabetic kidney disease type 2, gestational diabetes mellitus, aging process, Inflammatory factors in petrochemical workers, acute necrotizing pancreatitis, and polycystic ovary syndrome. But, no significant effect was reported on the oxidative status in patients with Rheumatoid Arthritis. Conclusions: Probiotics supplements could improvement oxidative stress biomarkers in various diseases. There are still some controversies about these methods and types of probiotics supplementations. So, it is recommended that more studies should be conducted for a better decision.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47701947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-09-27DOI: 10.30491/JABR.2020.114985
Mahmoud Gharbavi, A. Sharafi, P. Fath, Saeed Oruji, H. Pakzad, H. Manjili
Introduction: Paclitaxel (PTX), an effective chemotherapeutic drug, is widely used to treat several types of cancers. However, its use is limited due to poor water solubility resulted in poor bioavailability. One of the main ways to increase the efficacy and endurance of medications depends on the carrier that used for it. In the current study, the microemulsions (ME) were investigated based on Poly [2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-nitrophenyl-oxycrabonyl poly ethylene glycol-methacrylate (ME-ONP)] (PMBN) ability as a carrier for PTX to increase the half-life and its bioavailability. Also, the physicochemical properties of the ME system were evaluated. Materials and Methods: PMBN, tween 80, triacetin, and glycerol were used as the drug carrier, surfactant, oil phase, and co-surfactant, respectively. The hemolytic activity was characterized using the RBC hemolysis assay to evaluate the blood compatibility of the MEs. In addition, the in vitro cytotoxic effect of PMBN-PTX-ME and PTX on MCF-7, 4T1, and HFF-2 cell lines was performed. PI and Annexin-V dyes were used for cell apoptosis. Results: The conductivity of ME evaluated and the results showed 432 to 589 µS/cm. In vitro, the drug release study revealed that PTX has controlled release at different pH levels. Refractive Index (RI) and % transmittance of the MEs remained ranging from 1.43 to 1.53, and 89% to 98%, respectively. The MTT assay determined that PMBN-ME without PTX had no significant toxicity on HFF-2, MCF-7, and 4T-1 cell lines. Based on the apoptosis assay, treated cell lines with PMBN approximately remained alive. Conclusions: The results revealed that ME-based on PMBN would be a promising drug delivery system for PTX drug delivery.
{"title":"Formulation and Biocompatibility of Microemulsion-Based PMBN as an Efficient System for Paclitaxel Delivery","authors":"Mahmoud Gharbavi, A. Sharafi, P. Fath, Saeed Oruji, H. Pakzad, H. Manjili","doi":"10.30491/JABR.2020.114985","DOIUrl":"https://doi.org/10.30491/JABR.2020.114985","url":null,"abstract":"Introduction: Paclitaxel (PTX), an effective chemotherapeutic drug, is widely used to treat several types of cancers. However, its use is limited due to poor water solubility resulted in poor bioavailability. One of the main ways to increase the efficacy and endurance of medications depends on the carrier that used for it. In the current study, the microemulsions (ME) were investigated based on Poly [2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-nitrophenyl-oxycrabonyl poly ethylene glycol-methacrylate (ME-ONP)] (PMBN) ability as a carrier for PTX to increase the half-life and its bioavailability. Also, the physicochemical properties of the ME system were evaluated. \u0000Materials and Methods: PMBN, tween 80, triacetin, and glycerol were used as the drug carrier, surfactant, oil phase, and co-surfactant, respectively. The hemolytic activity was characterized using the RBC hemolysis assay to evaluate the blood compatibility of the MEs. In addition, the in vitro cytotoxic effect of PMBN-PTX-ME and PTX on MCF-7, 4T1, and HFF-2 cell lines was performed. PI and Annexin-V dyes were used for cell apoptosis. Results: The conductivity of ME evaluated and the results showed 432 to 589 µS/cm. In vitro, the drug release study revealed that PTX has controlled release at different pH levels. Refractive Index (RI) and % transmittance of the MEs remained ranging from 1.43 to 1.53, and 89% to 98%, respectively. The MTT assay determined that PMBN-ME without PTX had no significant toxicity on HFF-2, MCF-7, and 4T-1 cell lines. Based on the apoptosis assay, treated cell lines with PMBN approximately remained alive. Conclusions: The results revealed that ME-based on PMBN would be a promising drug delivery system for PTX drug delivery.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46373667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-08-13DOI: 10.30491/JABR.2020.117881
A. Pal, A. Paul
Introduction: Microbial endophytes colonizing internal tissues of living plants provide benefits to their host by promoting plant growth and protection against microbial infectious through the production of wide ranges of metabolites. Members of such endophytic community harbouring medicinally important plants are known to synthesize several antimicrobial compounds. The present study aims to explore bacterial endophytes of ethnomedicinal plant Rauvolfia serpentina (Apocynaceae) for producing novel antimicrobial metabolites of pharmaceutical and biotechnological importance. Materials and Methods: The culturable bacterial endophytic diversity of R. serpentina (L.) Benth. ex. Kurz. has been screened for producing antimicrobial compounds following cross-streak and agar well diffusion assay methods against several test microbial strains. The bioactive compound was isolated and partially purified from the cell-free culture filtrate following chromatographic methods. Results: The endophytes revealed low colonization frequency and isolation rates in the root and stem respectively. In vitro antimicrobial screening of 12 phenotypically distinguishable endophytes resulted in the selection of a potent antimicrobial isolate RAU 305 identified as Pseudomonas aeruginosa RAU 305 (Genbank accession number KR816098). Cell-free culture filtrate of RAU 305 showed broad spectrum of antimicrobial activity by inhibiting Paenibacillus, Micrococcus, Arthrobacter, Rhodobacter, Mycobacterium, Bacillus, Escherichia, Staphylococcus, Klebsiella, Aspergillus, Colletotrichum and Pythium. The antimicrobially active component extracted in butanol and chloroform was partially purified by column chromatography followed by a preparative thin layer chromatography and the homogeneity of the compound was confirmed using different solvent systems. Conclusions: More detailed characterization and identification of the active component is essential to explore the metabolic potential of this endophytic bacterium in future.
{"title":"In Vitro Antimicrobial Activity Screening of Bacteria Endophytic to Ethnomedicinal Plant Rauvolfia serpentina (L.) Benth. ex. Kurz","authors":"A. Pal, A. Paul","doi":"10.30491/JABR.2020.117881","DOIUrl":"https://doi.org/10.30491/JABR.2020.117881","url":null,"abstract":"Introduction: Microbial endophytes colonizing internal tissues of living plants provide benefits to their host by promoting plant growth and protection against microbial infectious through the production of wide ranges of metabolites. Members of such endophytic community harbouring medicinally important plants are known to synthesize several antimicrobial compounds. The present study aims to explore bacterial endophytes of ethnomedicinal plant Rauvolfia serpentina (Apocynaceae) for producing novel antimicrobial metabolites of pharmaceutical and biotechnological importance. Materials and Methods: The culturable bacterial endophytic diversity of R. serpentina (L.) Benth. ex. Kurz. has been screened for producing antimicrobial compounds following cross-streak and agar well diffusion assay methods against several test microbial strains. The bioactive compound was isolated and partially purified from the cell-free culture filtrate following chromatographic methods. Results: The endophytes revealed low colonization frequency and isolation rates in the root and stem respectively. In vitro antimicrobial screening of 12 phenotypically distinguishable endophytes resulted in the selection of a potent antimicrobial isolate RAU 305 identified as Pseudomonas aeruginosa RAU 305 (Genbank accession number KR816098). Cell-free culture filtrate of RAU 305 showed broad spectrum of antimicrobial activity by inhibiting Paenibacillus, Micrococcus, Arthrobacter, Rhodobacter, Mycobacterium, Bacillus, Escherichia, Staphylococcus, Klebsiella, Aspergillus, Colletotrichum and Pythium. The antimicrobially active component extracted in butanol and chloroform was partially purified by column chromatography followed by a preparative thin layer chromatography and the homogeneity of the compound was confirmed using different solvent systems. Conclusions: More detailed characterization and identification of the active component is essential to explore the metabolic potential of this endophytic bacterium in future.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"176-184"},"PeriodicalIF":0.0,"publicationDate":"2020-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48234049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-07-28DOI: 10.30491/JABR.2020.111083
Mojtaba Sharti, H. Ghaleh, R. Dorostkar, B. Kondori
Introduction: Antiviral therapy is an alternative for viral infection control when the virus is identified. As antiviral therapy has effectively used basic science to create very efficient treatments for severe viral infections, it is one of the most promising virology aspects. In the present work, a novel broad-spectrum antiviral method, dubbed Double-stranded RNA (dsRNA) Activated Caspase Oligomerizer (DRACO) have been developed, which induces apoptosis in cells with viral dsRNA selectively to kill infected cells quickly with no damage to uninfected ones. Materials and Methods: Following the design, development, expression, and purification of DRACO, influenza virus-infected MDCK and uninfected MDCK cells were treated with 40, 60, and 80 mg/L concentration of DRACO to study its potential antiviral activity. Then, TCID50 (50% Tissue Culture Infectious Dose) of the virus, together with the viability of cells, was measured. Results: The findings of the present study showed that DRACO is nontoxic to uninfected MDCK cells and is effective for H1N1 influenza virus-infected MDCK cells dose-dependently. Also, the infected MDCK cells treated with DRACO have shown a significant reduction in TCID50 compared with the control group. Conclusions: The outcomes suggest that DRACO has potential as a new anti-H1N1 therapeutic drug that its in-vivo antiviral efficacy requires to be examined through a clinical analysis of large quantities of animal models.
{"title":"Double-Stranded RNA Activated Caspase Oligomerizer (DRACO): Design, Subcloning, and Antiviral Investigation","authors":"Mojtaba Sharti, H. Ghaleh, R. Dorostkar, B. Kondori","doi":"10.30491/JABR.2020.111083","DOIUrl":"https://doi.org/10.30491/JABR.2020.111083","url":null,"abstract":"Introduction: Antiviral therapy is an alternative for viral infection control when the virus is identified. As antiviral therapy has effectively used basic science to create very efficient treatments for severe viral infections, it is one of the most promising virology aspects. In the present work, a novel broad-spectrum antiviral method, dubbed Double-stranded RNA (dsRNA) Activated Caspase Oligomerizer (DRACO) have been developed, which induces apoptosis in cells with viral dsRNA selectively to kill infected cells quickly with no damage to uninfected ones. Materials and Methods: Following the design, development, expression, and purification of DRACO, influenza virus-infected MDCK and uninfected MDCK cells were treated with 40, 60, and 80 mg/L concentration of DRACO to study its potential antiviral activity. Then, TCID50 (50% Tissue Culture Infectious Dose) of the virus, together with the viability of cells, was measured. Results: The findings of the present study showed that DRACO is nontoxic to uninfected MDCK cells and is effective for H1N1 influenza virus-infected MDCK cells dose-dependently. Also, the infected MDCK cells treated with DRACO have shown a significant reduction in TCID50 compared with the control group. Conclusions: The outcomes suggest that DRACO has potential as a new anti-H1N1 therapeutic drug that its in-vivo antiviral efficacy requires to be examined through a clinical analysis of large quantities of animal models.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43693596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-07-13DOI: 10.30491/JABR.2020.117885
M. Hosseini, M. Ebrahimi, E. Salehghamari, Amir Salehi Najafabadi, B. Yakhchali
Introduction: Biotransformation process has been used in various industries due to its ability to produce valuable chemicals and address environmental concerns. Propylene hydroformylation is a process in which n-butyraldehyde and isobutyraldehyde are produced. N-butyraldehyde is a high valuable chemical with many industrial applications, while isobutyraldehyde produced as a by-product is an environmental pollutant. This study offers a biotechnological approach for conversion of isobutyraldehyde into a high-value substance. An engineered strain of Escherichia coli was developed by genomic insertion of alcohol-dehydrogenase gene (adhA) from Lactococcus lactis which can convert isobutyraldehyde into isobutanol. Materials and Methods: The adhA gene was engineered to substitute some of its amino acids to result in a more efficient enzyme. Engineered gene was synthesized and introduced into E. coli genome to develop recombinant E. coli EG-296 strain. In addition, by using the Qualiteck-4 software, 16 well-defined experiments (L16 Orthogonal array) with two levels of seven variable parameters were used to optimize the process efficiency. Results: The findings of this study revealed that the E. coli strain EG-296 is capable of converting isobutyraldehyde into isobutanol. The optimization results showed that optimum medium composition for the highest isobutanol production were 10 g/L glucose or glycerol as carbon source, 10 g/L NH4CL as nitrogen source, mid-log of inoculum age, and 1% inoculum volume in 25ml medium. After optimization, 560 mg/L isobutanol was produced from 600 mg/L isobutyraldehyde with 91% yield. Conclusions: Recombinant E. coli strain with a relatively optimum medium can be used to remove isobutyraldehyde in refineries or other industries producing this chemical as a by-product.
{"title":"Biotransformation of Isobutyraldehyde to Isobutanol by an Engineered Escherichia coli Strain","authors":"M. Hosseini, M. Ebrahimi, E. Salehghamari, Amir Salehi Najafabadi, B. Yakhchali","doi":"10.30491/JABR.2020.117885","DOIUrl":"https://doi.org/10.30491/JABR.2020.117885","url":null,"abstract":"Introduction: Biotransformation process has been used in various industries due to its ability to produce valuable chemicals and address environmental concerns. Propylene hydroformylation is a process in which n-butyraldehyde and isobutyraldehyde are produced. N-butyraldehyde is a high valuable chemical with many industrial applications, while isobutyraldehyde produced as a by-product is an environmental pollutant. This study offers a biotechnological approach for conversion of isobutyraldehyde into a high-value substance. An engineered strain of Escherichia coli was developed by genomic insertion of alcohol-dehydrogenase gene (adhA) from Lactococcus lactis which can convert isobutyraldehyde into isobutanol. Materials and Methods: The adhA gene was engineered to substitute some of its amino acids to result in a more efficient enzyme. Engineered gene was synthesized and introduced into E. coli genome to develop recombinant E. coli EG-296 strain. In addition, by using the Qualiteck-4 software, 16 well-defined experiments (L16 Orthogonal array) with two levels of seven variable parameters were used to optimize the process efficiency. Results: The findings of this study revealed that the E. coli strain EG-296 is capable of converting isobutyraldehyde into isobutanol. The optimization results showed that optimum medium composition for the highest isobutanol production were 10 g/L glucose or glycerol as carbon source, 10 g/L NH4CL as nitrogen source, mid-log of inoculum age, and 1% inoculum volume in 25ml medium. After optimization, 560 mg/L isobutanol was produced from 600 mg/L isobutyraldehyde with 91% yield. Conclusions: Recombinant E. coli strain with a relatively optimum medium can be used to remove isobutyraldehyde in refineries or other industries producing this chemical as a by-product.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"159-165"},"PeriodicalIF":0.0,"publicationDate":"2020-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47223574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-07-12DOI: 10.30491/JABR.2020.110332
J. Abraham, A. Chatterjee, J. Sharma
Introduction: Increased usage and improper management of electronic wastes result in immense environmental pollution. Although conventional techniques are well known for heavy metals removal from the environment, their high cost and severe environmental consequences indicate the urgent requirement of cost-effective methods of heavy metals uptake. Bioaccumulation can be considered as an alternative to the traditional methods in terms of their cost-effectiveness and maximum recovery of the metal ions. Materials and Methods: This study deals with the isolation of heavy metals tolerant Gram-positive bacterial strain, Bacillus licheniformis JAJ3, and its application in bioaccumulation of copper, lead, and nickel and bioleaching of heavy metals from electronic waste. 16S rRNA sequencing was performed to identify the bacterial strain. The accumulation study was carried out in a liquid medium and analyzed using atomic absorption spectroscopy. Bioleaching activity was checked using the one-step procedure. For bioleaching studies of heavy metals, printed circuit boards (PCBs) were used as a source of electronic wastes. Scanning electron microscopy and energy dispersive spectroscopy were used to record the changes before and after experimental procedures. Results: The organism was able to accumulate 98.6% copper, 64.6% lead, and 57.3% nickel. The bioaccumulation reaction followed pseudo-second order kinetics model (R2 value 0.92, 0.92, 0.99 for copper, lead, and nickel bioaccumulation respectively). Efficient bioleaching activity was shown by the strain. Conclusions: The experimental analyses confirmed that the strain is efficient in the bioleaching of heavy metals from electronic wastes and thus can be used in management of the electronic wastes.
{"title":"Isolation and Characterization of Bacillus licheniformis Strain for Bioleaching of Heavy Metals","authors":"J. Abraham, A. Chatterjee, J. Sharma","doi":"10.30491/JABR.2020.110332","DOIUrl":"https://doi.org/10.30491/JABR.2020.110332","url":null,"abstract":"Introduction: Increased usage and improper management of electronic wastes result in immense environmental pollution. Although conventional techniques are well known for heavy metals removal from the environment, their high cost and severe environmental consequences indicate the urgent requirement of cost-effective methods of heavy metals uptake. Bioaccumulation can be considered as an alternative to the traditional methods in terms of their cost-effectiveness and maximum recovery of the metal ions. Materials and Methods: This study deals with the isolation of heavy metals tolerant Gram-positive bacterial strain, Bacillus licheniformis JAJ3, and its application in bioaccumulation of copper, lead, and nickel and bioleaching of heavy metals from electronic waste. 16S rRNA sequencing was performed to identify the bacterial strain. The accumulation study was carried out in a liquid medium and analyzed using atomic absorption spectroscopy. Bioleaching activity was checked using the one-step procedure. For bioleaching studies of heavy metals, printed circuit boards (PCBs) were used as a source of electronic wastes. Scanning electron microscopy and energy dispersive spectroscopy were used to record the changes before and after experimental procedures. Results: The organism was able to accumulate 98.6% copper, 64.6% lead, and 57.3% nickel. The bioaccumulation reaction followed pseudo-second order kinetics model (R2 value 0.92, 0.92, 0.99 for copper, lead, and nickel bioaccumulation respectively). Efficient bioleaching activity was shown by the strain. Conclusions: The experimental analyses confirmed that the strain is efficient in the bioleaching of heavy metals from electronic wastes and thus can be used in management of the electronic wastes.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"139-144"},"PeriodicalIF":0.0,"publicationDate":"2020-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49657501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-07-11DOI: 10.30491/JABR.2020.224143.1201
P. Agre, C. Nwachukwu, B. Olasanmi, J. Obidiegwu, E. Nwachukwu, P. Adebola, D. DeKoeyer, A. Asrat
Introduction: The methods that reliably yield quality DNA and distinguishing sex type at the early stage of growth have been a challenge in yam genetics and breeding studies. This study assessed the effect of sample preservation methods on DNA quantity and quality during extraction and potential of DNA marker to diagnose plant sex at the early seedling stage in white Guinea yam. Materials and Methods: Five sample preservation methods were assessed for quality DNA extraction during field leaf tissue collection, namely liquid nitrogen, dry ice, silica gel, 95% ethanol, and oven drying. The predicted sex at the seedling stage using the molecular marker was further validated with the visual score for the sex phenotype at the flowering stage. Results: According to the findings of the present study, the DNA extracted from leaf samples preserved in liquid nitrogen, silica gel, dry ice, and oven drying methods were higher in molecular weights than samples stored in ethanol solution. Yam plant sex diagnosis with the DNA marker (sp16) identified a higher proportion of ZW genotypes (female or monoecious phenotypes) than the ZZ genotypes (male phenotypes) in the studied materials with 74% prediction accuracy. Conclusions: The results from this study provided valuable insights on suitable sample preservation methods for quality DNA extraction and the potential of DNA marker sp16 to predict sex in white Guinea yam.
{"title":"Sample Preservation and Plant Sex Prediction in White Guinea yam (Dioscorea rotundata Poir.)","authors":"P. Agre, C. Nwachukwu, B. Olasanmi, J. Obidiegwu, E. Nwachukwu, P. Adebola, D. DeKoeyer, A. Asrat","doi":"10.30491/JABR.2020.224143.1201","DOIUrl":"https://doi.org/10.30491/JABR.2020.224143.1201","url":null,"abstract":"Introduction: The methods that reliably yield quality DNA and distinguishing sex type at the early stage of growth have been a challenge in yam genetics and breeding studies. This study assessed the effect of sample preservation methods on DNA quantity and quality during extraction and potential of DNA marker to diagnose plant sex at the early seedling stage in white Guinea yam. Materials and Methods: Five sample preservation methods were assessed for quality DNA extraction during field leaf tissue collection, namely liquid nitrogen, dry ice, silica gel, 95% ethanol, and oven drying. The predicted sex at the seedling stage using the molecular marker was further validated with the visual score for the sex phenotype at the flowering stage. Results: According to the findings of the present study, the DNA extracted from leaf samples preserved in liquid nitrogen, silica gel, dry ice, and oven drying methods were higher in molecular weights than samples stored in ethanol solution. Yam plant sex diagnosis with the DNA marker (sp16) identified a higher proportion of ZW genotypes (female or monoecious phenotypes) than the ZZ genotypes (male phenotypes) in the studied materials with 74% prediction accuracy. Conclusions: The results from this study provided valuable insights on suitable sample preservation methods for quality DNA extraction and the potential of DNA marker sp16 to predict sex in white Guinea yam.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45006277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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