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}
Pub Date : 2020-07-11DOI: 10.30491/JABR.2020.110243
R. Sarvestani, A. Latifi, H. Alizadeh, M. Mirzaei
Introduction: Wide applications in research, clinical and cosmetic industry of human epidermal growth factor (hEGF) made it a research interest target. Its production in different expression systems has shown several limitations. Recombinant expression of hEGF in E. coli is always accompanied by inclusion body formation. The object of this study is to the evaluation of a chromatography-independent approach for the production of EGF in E. coli as soluble form. Materials and Methods: In order to evaluate a chromatogram independent purification approach for recombinant hEGF production in a soluble form, the hEGF gene was fused to an elastin-like protein (ELP) and expressed in E. coli BL21 (DE3) using pET26b expression vector for secretion the product into periplasmic space. Results: Periplasmic protein content analysis confirmed that the recombinant protein is secreted into the periplasm. The purification process was done by using 0.4 M ammonium sulfate in two cycles of inverse phase transition (ITC). After two cycles of purification, purity reached more than 95%. Western blotting analysis with the monoclonal anti-EGF antibody has confirmed the accuracy of EGF. Biological activity of the purified protein was investigated on NIH-3T3 cell line and results indicated EGF-induced proliferation in treated cells. Our results showed periplasmic expression is the proper approach to the production of soluble recombinant hEGF. By using ELP fused to EGF, the purification process was established without applying chromatography which will result in decreasing in final costs. Conclusions: This study introduced a new economic and efficient approach to the production and purification of recombinant hEGF.
{"title":"An Approach for Recombinant Epidermal Growth Factor Purification by Using an Elastin-Like Protein Tag","authors":"R. Sarvestani, A. Latifi, H. Alizadeh, M. Mirzaei","doi":"10.30491/JABR.2020.110243","DOIUrl":"https://doi.org/10.30491/JABR.2020.110243","url":null,"abstract":"Introduction: Wide applications in research, clinical and cosmetic industry of human epidermal growth factor (hEGF) made it a research interest target. Its production in different expression systems has shown several limitations. Recombinant expression of hEGF in E. coli is always accompanied by inclusion body formation. The object of this study is to the evaluation of a chromatography-independent approach for the production of EGF in E. coli as soluble form. Materials and Methods: In order to evaluate a chromatogram independent purification approach for recombinant hEGF production in a soluble form, the hEGF gene was fused to an elastin-like protein (ELP) and expressed in E. coli BL21 (DE3) using pET26b expression vector for secretion the product into periplasmic space. Results: Periplasmic protein content analysis confirmed that the recombinant protein is secreted into the periplasm. The purification process was done by using 0.4 M ammonium sulfate in two cycles of inverse phase transition (ITC). After two cycles of purification, purity reached more than 95%. Western blotting analysis with the monoclonal anti-EGF antibody has confirmed the accuracy of EGF. Biological activity of the purified protein was investigated on NIH-3T3 cell line and results indicated EGF-induced proliferation in treated cells. Our results showed periplasmic expression is the proper approach to the production of soluble recombinant hEGF. By using ELP fused to EGF, the purification process was established without applying chromatography which will result in decreasing in final costs. Conclusions: This study introduced a new economic and efficient approach to the production and purification of recombinant hEGF.","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":"48575563","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 : 2020-07-07DOI: 10.30491/JABR.2020.109997
Zhi Xin Phuna, James Ken Ee Yu, Jie Ying Tee, S. Chuah, N. Tan, S. Vijayabalan, A. Manap, Sreenivas Patro Sisinthy, P. Madhavan
Introduction: Vulvovaginal candidiasis (VVC) has received enormous attention, not only due to its negative influences on women’s life but also because of the escalating trend of fungal resistance towards current antifungal drugs. In recent decades, researches have been focusing on the development of natural products as the antifungal agents due to their low side effects compared to standard antifungal drugs. In this study, the antifungal activity of curcumin, piperine, and tualang honey (TH) in single, combination, and combined nanoemulsions was evaluated. Materials and Methods: The nanoemulsions were prepared by dissolving curcumin, piperine and TH in the nanoemulsions base which was prepared by mixing Capryol, Tween80/Kolliphor RN40 and Transcutol HP. For assesment of atifungal activity, well diffusion methods were used and the zone of inhibitions were compared to fluconazole as a standard drug, . Results: The antifungal activity of these natural products alone was low and not all combinations were significant. Moreover, both curcumin and piperine are known to have low bioavailability that might limit its fungicidal efficiency. Hence, nanoemulsions of curcumin, piperine, and honey were then developed in this study. The nanoemulsions of three natural compounds have possessed favorable antifungal activity (more than 80%) against the wide range of Candida spp. Particularly, Candida albicans was more susceptible to these nanoemulsions compared to other species tested and some of them were the most resistant to fluconazole. Conclusions: In concise, this study showed evidence in support of the therapeutic use of nanoemulsions of curcumin, piperine, and tualang honey in antifungal infections.
{"title":"In Vitro Evaluation of Nanoemulsions of Curcumin, Piperine, and Tualang Honey as Antifungal Agents for Candida Species","authors":"Zhi Xin Phuna, James Ken Ee Yu, Jie Ying Tee, S. Chuah, N. Tan, S. Vijayabalan, A. Manap, Sreenivas Patro Sisinthy, P. Madhavan","doi":"10.30491/JABR.2020.109997","DOIUrl":"https://doi.org/10.30491/JABR.2020.109997","url":null,"abstract":"Introduction: Vulvovaginal candidiasis (VVC) has received enormous attention, not only due to its negative influences on women’s life but also because of the escalating trend of fungal resistance towards current antifungal drugs. In recent decades, researches have been focusing on the development of natural products as the antifungal agents due to their low side effects compared to standard antifungal drugs. In this study, the antifungal activity of curcumin, piperine, and tualang honey (TH) in single, combination, and combined nanoemulsions was evaluated. Materials and Methods: The nanoemulsions were prepared by dissolving curcumin, piperine and TH in the nanoemulsions base which was prepared by mixing Capryol, Tween80/Kolliphor RN40 and Transcutol HP. For assesment of atifungal activity, well diffusion methods were used and the zone of inhibitions were compared to fluconazole as a standard drug, . Results: The antifungal activity of these natural products alone was low and not all combinations were significant. Moreover, both curcumin and piperine are known to have low bioavailability that might limit its fungicidal efficiency. Hence, nanoemulsions of curcumin, piperine, and honey were then developed in this study. The nanoemulsions of three natural compounds have possessed favorable antifungal activity (more than 80%) against the wide range of Candida spp. Particularly, Candida albicans was more susceptible to these nanoemulsions compared to other species tested and some of them were the most resistant to fluconazole. Conclusions: In concise, this study showed evidence in support of the therapeutic use of nanoemulsions of curcumin, piperine, and tualang honey in antifungal infections.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"189-197"},"PeriodicalIF":0.0,"publicationDate":"2020-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43913358","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 : 2020-07-07DOI: 10.30491/JABR.2020.109994
K. Alam, Muhammad Iqbal, A. Hasan, N. Al-Maskari
Introduction: Biological hydrogels provide a conducive extracellular environment for encapsulating and growing cells and play an important role in regulating cell behavior. Mechanical and rheological properties of hydrogels can influence cell function, mechanotransduction and cellular behaviors such as growth, migration, adhesion, self-renewal, differentiation, morphology and fate. Determination of rheological properties of biogels is important for printing tissues by controlling physical properties and developing efficient drug delivery systems. The main purpose of the current study was to determine some important rheological properties of two well-known hydrogels (agarose and gelatin methacryloyl [GelMA]). Materials and Methods: Rheological properties of gel solutions with different concentrations were measured using oscillatory rheometry. Agarose gels of 1% and 2% (w/v) concentration were prepared in 100 mL de-ionized water. The GelMA solutions of 10% and 15% concentrations were prepared by dissolving dry GelMA in deionized water. Rheological measurements were performed using a rheometer with cone-plate geometry. Results: Both storage modulus (G′) and loss modulus (G′′) increased with an increase in frequency. Rheological properties of both types of gel solutions were strongly influenced by the amount of concentration. The shear stress profiles demonstrated shear thinning in both types of gels. Viscosity of 1% agarose and 2% agarose was found comparable with 10% GelMA and 15% GelMA , respectively. Conclusions: Results obtained from experiments revealed that rotational rheometry can be confidently used to determine viscous and elastic response of hydrogels in the aqueous state. The results will help to select the right type of gel and amount of concentration for the bio-printing of tissues.
{"title":"Rheological characterization of biological hydrogels in aqueous state","authors":"K. Alam, Muhammad Iqbal, A. Hasan, N. Al-Maskari","doi":"10.30491/JABR.2020.109994","DOIUrl":"https://doi.org/10.30491/JABR.2020.109994","url":null,"abstract":"Introduction: Biological hydrogels provide a conducive extracellular environment for encapsulating and growing cells and play an important role in regulating cell behavior. Mechanical and rheological properties of hydrogels can influence cell function, mechanotransduction and cellular behaviors such as growth, migration, adhesion, self-renewal, differentiation, morphology and fate. Determination of rheological properties of biogels is important for printing tissues by controlling physical properties and developing efficient drug delivery systems. The main purpose of the current study was to determine some important rheological properties of two well-known hydrogels (agarose and gelatin methacryloyl [GelMA]). Materials and Methods: Rheological properties of gel solutions with different concentrations were measured using oscillatory rheometry. Agarose gels of 1% and 2% (w/v) concentration were prepared in 100 mL de-ionized water. The GelMA solutions of 10% and 15% concentrations were prepared by dissolving dry GelMA in deionized water. Rheological measurements were performed using a rheometer with cone-plate geometry. Results: Both storage modulus (G′) and loss modulus (G′′) increased with an increase in frequency. Rheological properties of both types of gel solutions were strongly influenced by the amount of concentration. The shear stress profiles demonstrated shear thinning in both types of gels. Viscosity of 1% agarose and 2% agarose was found comparable with 10% GelMA and 15% GelMA , respectively. Conclusions: Results obtained from experiments revealed that rotational rheometry can be confidently used to determine viscous and elastic response of hydrogels in the aqueous state. The results will help to select the right type of gel and amount of concentration for the bio-printing of tissues.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"171-175"},"PeriodicalIF":0.0,"publicationDate":"2020-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43530834","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 : 2020-07-07DOI: 10.30491/JABR.2020.109990
Lutfun Neesa, R. Islam, N. Jahan, U. S. Zohora, Mohammad Shahedur Rahman
Introduction: The increased need for a considerable β-glucosidase activity, especially in the enzymatic saccharification of cellulose for bioenergy, has strongly stimulated the identification of effective β-glucosidase producing microbes. This study was conducted to optimize culture condition for β-glucosidase production from the identified new isolate of Bacillus subtilis (B1) and to find out an ideal condition for β-glucosidase activity. Materials and Methods: For β-glucosidase production, the bacterium was cultivated in a basal medium. The culture condition was optimized at several pH, different temperatures, varying cultivation periods, and various substrate concentrations. Finally, the activity of the β-glucosidase enzyme was investigated at different incubation periods, pH, temperatures, metal ions, and various percentages of methanol. The activity of β-glucosidase was measured by the capability of crude enzyme to convert pNPG (p-nitrophenyl-β-D glucopyranoside) into yellow product PNP (p-nitrophenol). Results: Cellulolytic bacterial strain B. subtilis (B1) showed high potentiality for β-glucosidase production at a pH of 7.0 after 24 hours incubation at 40°C. The highest level of enzyme production was achieved when 3% of CMC was provided in the culture medium. Optimum reaction conditions for β-glucosidase activity were shown to be 10 minutes, 60°C and at pH 7. Salts like Magnesium Sulfate (MgSO4), Calcium Chloride (CaCl2), and Manganese Sulfate (MnSO4) positively influenced the activity where NaCl and KCl had negative effects. The presence of methanol (80%) appreciably enhanced the activity of enzyme. Conclusions: Complete saccharification of different industrial processes can be augmented by using this novel β-glucosidase produced by B. subtilis strain isolated from effluent of biogas plant.
{"title":"Optimization of Culture Conditions and Reaction Parameters of β-glucosidase From a New Isolate of Bacillus subtilis (B1)","authors":"Lutfun Neesa, R. Islam, N. Jahan, U. S. Zohora, Mohammad Shahedur Rahman","doi":"10.30491/JABR.2020.109990","DOIUrl":"https://doi.org/10.30491/JABR.2020.109990","url":null,"abstract":"Introduction: The increased need for a considerable β-glucosidase activity, especially in the enzymatic saccharification of cellulose for bioenergy, has strongly stimulated the identification of effective β-glucosidase producing microbes. This study was conducted to optimize culture condition for β-glucosidase production from the identified new isolate of Bacillus subtilis (B1) and to find out an ideal condition for β-glucosidase activity. \u0000Materials and Methods: For β-glucosidase production, the bacterium was cultivated in a basal medium. The culture condition was optimized at several pH, different temperatures, varying cultivation periods, and various substrate concentrations. Finally, the activity of the β-glucosidase enzyme was investigated at different incubation periods, pH, temperatures, metal ions, and various percentages of methanol. The activity of β-glucosidase was measured by the capability of crude enzyme to convert pNPG (p-nitrophenyl-β-D glucopyranoside) into yellow product PNP (p-nitrophenol). \u0000Results: Cellulolytic bacterial strain B. subtilis (B1) showed high potentiality for β-glucosidase production at a pH of 7.0 after 24 hours incubation at 40°C. The highest level of enzyme production was achieved when 3% of CMC was provided in the culture medium. Optimum reaction conditions for β-glucosidase activity were shown to be 10 minutes, 60°C and at pH 7. Salts like Magnesium Sulfate (MgSO4), Calcium Chloride (CaCl2), and Manganese Sulfate (MnSO4) positively influenced the activity where NaCl and KCl had negative effects. The presence of methanol (80%) appreciably enhanced the activity of enzyme. \u0000Conclusions: Complete saccharification of different industrial processes can be augmented by using this novel β-glucosidase produced by B. subtilis strain isolated from effluent of biogas plant.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42129631","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 : 2020-07-05DOI: 10.30491/JABR.2020.109897
Soumia Attou, B. Meddah, A. T. Meddah, Meriem Mokhtar, P. Sonnet
Introduction: Aristolochia longa, is widely used as a medicinal plant, in Algerian folk medicine since ancient times and passed through the generations. This study focused on the qualitative examination of different phytochemical constituents to determine their antioxidant activities.Materials and Methods: Leaves and roots were extracted by liquid-liquid extraction technique using the methanol as solvent. The total phenolic and flavonoids content was determined by Folin–Ciocalteu and AlCl3 methods, respectively. The antioxidant activity was evaluated with two distinct methods: DPPH radical scavenging assay and ferric reducing antioxidant power test (FRAP). Then, the high-performance liquid chromatography (HPLC) method was performed to analyze leaves and roots n-butanol fractions. Results: Leaves gave a significant value of polyphenols (8.580 ± 0.04 mg GAE/g DW). Whereas, n-butanol fraction flavonoids extracted from leaves was observed much highest (4.54 ± 1.94 mg CE/g DW). N-butanol fraction leaves showed a powerful scavenging activity and reducing activity with an IC50 = 0.044 ± 0.001 mg/ml and EC50 = 0.126 ± 0.041 mg/ml, respectively. The HPLC analysis of leaves and roots n-butanol fraction revealed different bioactive compounds in which they belong to the flavonoids category.Conclusions: The results obtained from this study suggest that Aristolochia longa leaves were considered as an important resource of flavonoids, which have an interesting antioxidant power.
{"title":"Phytochemical Screening and Antioxidant Activity of Algerian Aristolochia longa Flavonoids","authors":"Soumia Attou, B. Meddah, A. T. Meddah, Meriem Mokhtar, P. Sonnet","doi":"10.30491/JABR.2020.109897","DOIUrl":"https://doi.org/10.30491/JABR.2020.109897","url":null,"abstract":"Introduction: Aristolochia longa, is widely used as a medicinal plant, in Algerian folk medicine since ancient times and passed through the generations. This study focused on the qualitative examination of different phytochemical constituents to determine their antioxidant activities.Materials and Methods: Leaves and roots were extracted by liquid-liquid extraction technique using the methanol as solvent. The total phenolic and flavonoids content was determined by Folin–Ciocalteu and AlCl3 methods, respectively. The antioxidant activity was evaluated with two distinct methods: DPPH radical scavenging assay and ferric reducing antioxidant power test (FRAP). Then, the high-performance liquid chromatography (HPLC) method was performed to analyze leaves and roots n-butanol fractions. Results: Leaves gave a significant value of polyphenols (8.580 ± 0.04 mg GAE/g DW). Whereas, n-butanol fraction flavonoids extracted from leaves was observed much highest (4.54 ± 1.94 mg CE/g DW). N-butanol fraction leaves showed a powerful scavenging activity and reducing activity with an IC50 = 0.044 ± 0.001 mg/ml and EC50 = 0.126 ± 0.041 mg/ml, respectively. The HPLC analysis of leaves and roots n-butanol fraction revealed different bioactive compounds in which they belong to the flavonoids category.Conclusions: The results obtained from this study suggest that Aristolochia longa leaves were considered as an important resource of flavonoids, which have an interesting antioxidant power.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"166-170"},"PeriodicalIF":0.0,"publicationDate":"2020-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42579687","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 : 2020-07-04DOI: 10.30491/JABR.2020.109861
M. Sadeghi, Ali Didehban, A. Sharifi, E. Seyedrezazadeh
Introduction: MicroRNAs (miRNAs) play an important role in the expression of their target genes. The single-nucleotide polymorphisms (SNPs) in miRNAs may affect their function and expression. The aim of the present study was to investigate the association between miR-196a2 rs116614913 polymorphism and non-small cell lung cancer (NSCLC) in the Iranian population. Materials and Methods: This case-control study was performed among 103 lung cancer patients and 100 healthy controls. The polymerase chain-reaction restriction fragment length polymorphism (PCR-RFLP) method and direct sequencing were used for miR-196a2 polymorphism genotyping. Statistical analyses were performed using SPSS software and t test method. Results: According to the findings of this study, there was no significant association between rs11614913 polymorphisms and the risk of lung cancer in codominant model (CT vs. CC: OR = 0.67, TT vs. CC: OR = 0.74, CT + CC vs. CC: OR = 1.133), dominant model (CT+TT vs. CC: OR=0.657) and recessive model (TT vs. CC+CT: OR = 0.88). In addition, there was no relationship between the clinicopathological characteristics of patients and controls. Conclusions: In summary, findings indicated no significant association between miR-196a2 rs11614913 polymorphisms and lung cancer in the Iranian population. Further studies with larger sample sizes are recommended to verify these findings.
MicroRNAs (miRNAs)在靶基因的表达中起着重要的作用。mirna中的单核苷酸多态性(snp)可能影响其功能和表达。本研究的目的是研究伊朗人群中miR-196a2 rs116614913多态性与非小细胞肺癌(NSCLC)之间的关系。材料与方法:本研究在103例肺癌患者和100例健康对照者中进行。采用聚合酶链反应限制性片段长度多态性(PCR-RFLP)法和直接测序法对miR-196a2多态性进行基因分型。采用SPSS软件和t检验方法进行统计学分析。结果:本研究结果显示，共显性模型(CT vs. CC: OR= 0.67, TT vs. CC: OR= 0.74, CT+ CC vs. CC: OR= 1.133)、显性模型(CT+TT vs. CC: OR=0.657)和隐性模型(TT vs. CC+CT: OR= 0.88)中rs11614913多态性与肺癌发病风险无显著相关性。此外，患者的临床病理特征与对照组之间没有关系。结论:总之，研究结果表明miR-196a2 rs11614913多态性与伊朗人群肺癌之间无显著关联。建议采用更大样本量的进一步研究来验证这些发现。
{"title":"Investigation of the Association between a Genetic Variant in MiR-196a-2 Gene and the Risk of Lung Cancer in the Iranian Population","authors":"M. Sadeghi, Ali Didehban, A. Sharifi, E. Seyedrezazadeh","doi":"10.30491/JABR.2020.109861","DOIUrl":"https://doi.org/10.30491/JABR.2020.109861","url":null,"abstract":"Introduction: MicroRNAs (miRNAs) play an important role in the expression of their target genes. The single-nucleotide polymorphisms (SNPs) in miRNAs may affect their function and expression. The aim of the present study was to investigate the association between miR-196a2 rs116614913 polymorphism and non-small cell lung cancer (NSCLC) in the Iranian population. Materials and Methods: This case-control study was performed among 103 lung cancer patients and 100 healthy controls. The polymerase chain-reaction restriction fragment length polymorphism (PCR-RFLP) method and direct sequencing were used for miR-196a2 polymorphism genotyping. Statistical analyses were performed using SPSS software and t test method. Results: According to the findings of this study, there was no significant association between rs11614913 polymorphisms and the risk of lung cancer in codominant model (CT vs. CC: OR = 0.67, TT vs. CC: OR = 0.74, CT + CC vs. CC: OR = 1.133), dominant model (CT+TT vs. CC: OR=0.657) and recessive model (TT vs. CC+CT: OR = 0.88). In addition, there was no relationship between the clinicopathological characteristics of patients and controls. Conclusions: In summary, findings indicated no significant association between miR-196a2 rs11614913 polymorphisms and lung cancer in the Iranian population. Further studies with larger sample sizes are recommended to verify these findings.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"185-188"},"PeriodicalIF":0.0,"publicationDate":"2020-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43472784","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 : 2020-06-27DOI: 10.30491/JABR.2020.109498
B. Bakchiche, İlginç Kızılpınar Temizer, A. Güder, Ö. G. Çelemli, S. Ç. Yegin, S. Bardaweel, M. Ghareeb
Introduction: In the current study, the microscopic and chemical analysis of Algerian honey, pollen, and propolis were investigated. Materials and Methods: The chemical composition of the ethanolic extracts of honeybee products was determined via gas chromatography-mass spectrometry (GC-MS) analysis. Furthermore, their in vitro anticancer, antimicrobial, antioxidant activities, total phenolic content (TPC), and total flavonoid content (TFC) were evaluated. Anticancer activities were assessed using the MTT assay while the antimicrobial potential was studied using the microdilution method. The antioxidant activities were investigated using the 2,2’-diphenyl-1-picrylhydrazyl radical (DPPH), hydrogen peroxide scavenging activity (H2O2) and ferric reducing antioxidant power (FRAP). The TPC and TFC were evaluated via Folin-Ciocalteu’s and AlCl3 assays, respectively. Results: In the GC-MS analyses, 36 compounds were identified in the ethanol extract of pollen accounting for 92.73% of the total extract; linolenic acid was the most abundant compound (21.28%). Also, 23 compounds were identified in the ethanol extract of propolis representing 29.91% of the total extract; Z-nerolidol was the most abundant compound (8.96%). Moreover, 17 compounds were identified in the ethanol extract of honey representing 99.40% of the total extract while glyceraldehyde (27.07%) was the major abundant compound. The ethanol extract from pollen yielded the highest TPC with 1169.33 mg Gallic acid equivalent/g dry extract. In the DPPH assay, the SC50 values ranged from 50.74 to 53.05 μg/mL. Significant antimicrobial activities were associated with propolis with Gram-positive bacteria as the most sensitive microorganisms. In addition, remarkable anticancer activities were observed for propolis against five human cancer cell lines with LD50 values in the range of 3-160 μg/mL. Conclusions: Algerian Honeybee products, especially propolis, may be a potential source of naturally occurring bioactive compounds for the treatment of oxidative stress and cancer diseases.
{"title":"Chemical Composition and Biological Activities of Honeybee Products From Algeria","authors":"B. Bakchiche, İlginç Kızılpınar Temizer, A. Güder, Ö. G. Çelemli, S. Ç. Yegin, S. Bardaweel, M. Ghareeb","doi":"10.30491/JABR.2020.109498","DOIUrl":"https://doi.org/10.30491/JABR.2020.109498","url":null,"abstract":"Introduction: In the current study, the microscopic and chemical analysis of Algerian honey, pollen, and propolis were investigated. Materials and Methods: The chemical composition of the ethanolic extracts of honeybee products was determined via gas chromatography-mass spectrometry (GC-MS) analysis. Furthermore, their in vitro anticancer, antimicrobial, antioxidant activities, total phenolic content (TPC), and total flavonoid content (TFC) were evaluated. Anticancer activities were assessed using the MTT assay while the antimicrobial potential was studied using the microdilution method. The antioxidant activities were investigated using the 2,2’-diphenyl-1-picrylhydrazyl radical (DPPH), hydrogen peroxide scavenging activity (H2O2) and ferric reducing antioxidant power (FRAP). The TPC and TFC were evaluated via Folin-Ciocalteu’s and AlCl3 assays, respectively. Results: In the GC-MS analyses, 36 compounds were identified in the ethanol extract of pollen accounting for 92.73% of the total extract; linolenic acid was the most abundant compound (21.28%). Also, 23 compounds were identified in the ethanol extract of propolis representing 29.91% of the total extract; Z-nerolidol was the most abundant compound (8.96%). Moreover, 17 compounds were identified in the ethanol extract of honey representing 99.40% of the total extract while glyceraldehyde (27.07%) was the major abundant compound. The ethanol extract from pollen yielded the highest TPC with 1169.33 mg Gallic acid equivalent/g dry extract. In the DPPH assay, the SC50 values ranged from 50.74 to 53.05 μg/mL. Significant antimicrobial activities were associated with propolis with Gram-positive bacteria as the most sensitive microorganisms. In addition, remarkable anticancer activities were observed for propolis against five human cancer cell lines with LD50 values in the range of 3-160 μg/mL. Conclusions: Algerian Honeybee products, especially propolis, may be a potential source of naturally occurring bioactive compounds for the treatment of oxidative stress and cancer diseases.","PeriodicalId":14945,"journal":{"name":"Journal of Applied Biotechnology Reports","volume":"7 1","pages":"93-103"},"PeriodicalIF":0.0,"publicationDate":"2020-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48985790","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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