Karim Imangholiloo; Nasrin Moshtaghi; Seyed Hasan Marashi; Abdolreza Bagheri; Ahmad Sharifi
Abstract
Cellulose which is extremely produced by plants, can be used for biofuel production but this function needs chemical or enzymatic digestion. Cellulose hydrolysis of plant wastes for ethanol production requires a mixture of three enzyme groups, including endoglucanases, exoglucanases, and beta-glucosidases. ...
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Cellulose which is extremely produced by plants, can be used for biofuel production but this function needs chemical or enzymatic digestion. Cellulose hydrolysis of plant wastes for ethanol production requires a mixture of three enzyme groups, including endoglucanases, exoglucanases, and beta-glucosidases. The cellobiohydrolase enzyme (Cel6B) from Thermobifidia fusca has been used for cellulase activity extensively. This research aimed to express recombinant Cel6B enzyme in Pichia pastoris. For this purpose, cel6B gene in control of AOX1 promoter (methanol inducible) was introduced into Pichia pastoris. Amplification of cel6B gene was performed by PCR technique and then introduced into the Phil-S1 yeast vector. The recombinant construct contained the cel6B gene sequence and PHO1 signal sequence as secretion signal was transferred into Pichia pastoris GS115 strain. The transformed yeast cells expressed the recombinant Cel6B to yield 2.104 U (µmol/min)/ml of culture medium. Purified recombinant enzyme showed the best activity at 60 °C and pH 4.5 and this was agreed with optimum conditions for recombinant Cel6B enzymes which were produced in other systems. The purity of the enzyme was examined by SDS–PAGE technique, and a single band with a molecular weight about 59.6 kDa was observed. As cel6B gene sequence was not optimized for expression in the Pichia pastoris yeast, this could be one of the reasons for low level activity of recombinant Cel6B enzyme. This thermostable enzyme can be used for cellulolytic digestion of biomaterials in biofuel production research and other uses.
Azadeh Khadem; Nasrin Moshtaghi; Abdolreza Bagheri
Abstract
Somatic embryogenesis encompasses the same set of various developmental processes similar to zygotic embryogenesis. The conversion of somatic cells to embryos also requires stages of differentiation and reprogramming of cells. Since somatic embryogenesis is a complex process, a comprehensive investigation ...
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Somatic embryogenesis encompasses the same set of various developmental processes similar to zygotic embryogenesis. The conversion of somatic cells to embryos also requires stages of differentiation and reprogramming of cells. Since somatic embryogenesis is a complex process, a comprehensive investigation is required to identify the effective gene networks and their interactions with environmental factors. As part of this study, bioinformatics tools and molecular studies were used to gain a better understanding of Arabidopsis thaliana somatic embryogenesis. The enriched pathways of somatic embryogenesis and their core-enriched genes were identified using gene set enrichment analysis. The results indicated that significant interaction between hormones helps to induce and develop somatic embryos. The gene ontology (including biological process, molecular function and cellular compartment) of core-enriched genes revealed that lipid storage and metabolism as well as stress response are the active biological pathways during somatic embryogenesis. In the protein-protein interaction network, TIR1/AFBs as auxin receptors exhibited the greatest number of interactions and proteins involved in lipid storage and metabolism acted as mediators between auxin receptors and ethylene perception. Also, Kyoto encyclopedia of genes and genomes analysis indicated that the metabolism of membrane lipids during somatic embryogenesis of Arabidopsis is primarily related to the biosynthesis of jasmonates and their derivatives. This process is initiated by Lypooxygenase proteins in the chloroplast, while Acyl-CoA oxidase 1 (ACX1) and Oxophytodienoate reductase 3 (OPR3) proceed this process in the peroxisome. The qRT-PCR analysis also confirmed the role of these genes during somatic embryogenesis, as the activity of these genes decreased at the beginning of 2,4-D treatment, but it increased during somatic embryogenesis. According to these results, jasmonates play an important role during somatic embryogenesis by mediating auxin signaling and stress response.
Narges Fazili; Zahra-Soheila Soheili; Saeid Malekzadeh-Shafaroudi; Shahram Samiei; Shamila D.Alipoor; Nasrin Moshtaghi; Abouzar Bagheri
Abstract
Royal jelly (RJ) from queen honeybee larva as a traditional medicine agent has a variety of pharmacological benefits. In the present study, the effect of Royal jelly was investigated on the urinary bladder cancer cell line (HTB-9 5637). To determine the cell viability in different concentrations ...
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Royal jelly (RJ) from queen honeybee larva as a traditional medicine agent has a variety of pharmacological benefits. In the present study, the effect of Royal jelly was investigated on the urinary bladder cancer cell line (HTB-9 5637). To determine the cell viability in different concentrations of Royal jelly, MTT assay was performed. An in vitro wound healing assay was applied to investigate the effect of RJ on cell migration. The activity and gene expression level of matrix metalloproteinase 2 and 9 was assessed by zymography and Real time PCR respectively. R.J.S at the concentration of 0.7 mg/ml had a significant effect on reducing the proliferation rate of 5637 cells after 72h (p < 0.009). R.J.S significantly decreased cell migration and induced a significant decrease in the transcriptional level of MMP9 after 72h (0.5x; P < 0.049). However R.J.S did not impose any effect on the expression level and activity of matrix metalloproteinase 2. These results indicate the potential of RJ as a promised natural anti-proliferative and anti-metastatic drug in combination with advanced therapy methods for cancer treatment. Royal jelly has the potential to be more focused as an anti-metastatic drug to control tumor growth and can be considered as a more effective alternative to the current chemotherapy drugs.
Zohreh Sohrabi Nezhad; Hassan Marashi; Nasrin Moshtaghi
Abstract
Silver nanoparticles are widely used in manufacturing of different products considering their unique physical and chemical properties. Also they have been noticed in medical diagnosis and treatment because of their antibacterial properties. Physical and chemical methods of producing ...
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Silver nanoparticles are widely used in manufacturing of different products considering their unique physical and chemical properties. Also they have been noticed in medical diagnosis and treatment because of their antibacterial properties. Physical and chemical methods of producing nanoparticles, are expensive and are not safe enough as toxic substances may remain in the final preparations. To solve this problem, biological production of nanoparticles is considered as an efficient alternative method. In present study, synthesis of silver nanoparticles via seeds, petals, roots, and hairy root extracts of Calendula officinalis were performed. These nanoparticles were characterized by means of spectrophotometer, particle size analyzer and transmission electron microscope. Nanoparticles which were synthesized by hairy root extract showed the highest absorption at 430 nm (1/6 a.u) and the smallest size (5/3 nm), in comparison to other examined particles. Results confirmed the better performance of hairy root extracts in the synthesis of silver nanoparticles.
Najme Javanmardi; Abdolreza Bagheri; Nasrin Moshtaghi; Ahmad Sharifi; Abbas Hemati Kakhki
Abstract
Saffron (Crocus sativus) is the most valuable and indigenous crop in Iran. The stigmas of flower are used as
a popular natural flavouring, colouring and medicinal agent. However, the market suffers from frauds in this
plant such as mixing with safflower petals due to high profit. Identification of ...
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Saffron (Crocus sativus) is the most valuable and indigenous crop in Iran. The stigmas of flower are used as
a popular natural flavouring, colouring and medicinal agent. However, the market suffers from frauds in this
plant such as mixing with safflower petals due to high profit. Identification of these frauds with conventional
and biochemical methods is difficult and low sensitive. Therefore, application of molecular markers such as
random amplified polymorphic DNA (RAPD)/sequence characterized amplified regions (SCAR) is being
considered as an alternative. In this study, DNA was extracted from dry stigmas of 5 Saffron accessions and
dry petals of 7 safflower cultivars. RAPD reactions with ten 15-mer random primers resulted in two specific
monomorphic bands (500 and 700 bp) for safflower, while they were absent in saffron accessions. PCR
analysis with specific SCAR primers amplified two specific bands (414 and 589 bp) for safflowers in different
combinations of saffron stigmas and safflower petals. This was the case with very low rates or 1% of
safflower. Therefore, this method seems to be suitable for fraud identification of safflower petals in
commercial saffron samples.
