فهرست مطالب

Biomacromolecular Journal - Volume:6 Issue: 2, Autumn 2020
  • Volume:6 Issue: 2, Autumn 2020
  • تاریخ انتشار: 1400/08/30
  • تعداد عناوین: 10
  • Milad Rasuli, Nadia Fallah, Adeleh Divsalar * Pages 104-113
    Due to bacteria and viruses are the pathogens of many living organisms, it is important to introduce a novel modality to inactivate them. Each alternative platform should bring about an appropriate platform to overcome ongoing challenges such as the COVID-19 pandemic. Gas plasma as an emerging technology has recently become a promising solution to a number of challenges due to its diverse applications in healthcare and medicine, environmental remediation and pollution control, materials processing, electrochemistry, nanotechnology, etc. Based on the physical and chemical effects, plasma creates a unique opportunity for specific applications. This article first provides an overview of how gas plasma works and then describes its use in effectively inactivating bacteria and viruses. With reviewing recent works in mentioned areas, we emphasize the application of plasma for SARS-CoV-2 inactivation. Furthermore, we summarize insights regarding the inactivation mechanism of gas plasma.a a a a a a a a a a
    Keywords: Gas plasma, Plasma medicine, Virus inactivation, SARS-CoV-2 inactivation, Bacteria inactivation
  • Meimanat Nasiri, Mehran Arabi, Roohullah Hemmati *, Garshasb Rigi Pages 114-122
    Amylase catalyzes the hydrolysis of starch, glycogen and gives rise to certain products such as maltopentose, maltotetrose, maltotriose, maltose, and glucose. In the present study, earthworm Allolobophora choloretica, from Lumbricidae family, was used as animal model system. First, the earthworm cell extracts were precipitated by using a gradient of saturated ammonium sulfate and multi-step dialysis, and then α-amylase was purified by Amicon® Ultra filter. The purified enzyme was analyzed on SDS-PAGE and a single 37 KD band observed on the gel. Subsequently, optimum pH and optimum temperature of the purified α-amylase were estimated to be 7 and 53 ℃, respectively. Based on our results, at extreme acidic and alkaline pH conditions, the enzyme showed higher pH stability at pH 9 than 4. Moreover, the values of 〖∆H〗_D^#, 〖∆S〗_D^#and 〖∆G〗_D^# were 63.75 kcal.mol-1, 0.113 cal.mol-1K-1 and 26.66 kcal.mol-1, respectively. In conclusion, the purified moderately thermophilic amylase from earthworm Allolobophora choloretica can be exploited in different industries.
    Keywords: Amylase, Allolobophora choloretica, thermal stability, Purification
  • Bahareh Seifi Nahavandi, Maryam Ghobeh, Shahin Ahmadian, Azadeh Ebrahim Habibi, Parichehreh Yaghmaei * Pages 123-130

    Alzheimer's disease (AD) is characterized by amyloid plaques and neuronal death ultimately leading to dementia. Using natural therapies has always been a great concern for AD. Herein, Geraniol, as a natural monoterpene, was applied to examine its protective and therapeutic effects on a rat model of AD. In order to create Alzheimer’s rat model, bilateral injection of Amyloid β1–42 (Aβ1–42) was performed into rats’ hippocampus. Both therapeutic (post-AD induction) and protective effects of Geraniol consumption (100 mg/kg) were investigated on the antioxidant and brain histological parameters. In addition, Aβ1–42 peptide was driven toward fibril formation in vitro and effect of Geraniol (100 µM) was observed on Aβ1–42 fibrils. Alzheimer’s-induced group showed impairment in the antioxidant parameters along with loss of neuronal cells and amyloid plaque formation. Administration of Geraniol, in both treatment and protective modes, increased neurogenesis, reduced amyloid plaques, and improved antioxidant parameters. Therefore, Geraniol showed capability of improving AD signs as well as direct anti-fibril effect and it could be considered as neuroprotective.

    Keywords: Geraniol, Aβ1-42, Alzheimer’s Disease, Neurogenesis, Amyloid plaques
  • Sedigheh Khamehchian, Majid Tebianian *, Hossein Zolfagharian Pages 131-138
    An indirect ELISA for potency assessment of Naja naja oxiana horse antivenom was developed, with the aim of monitoring of antivenom immunoglobulin levels to replace the well-established conventional in vivo neutralization assay. According to this purpose, blood sera were taken during immunization schedule of the nineteen horses used in snake antivenom manufacture at the Razi Vaccine and Serum Research Institute. In addition, a group of fifteen batches of monovalent and polyvalent anti-snake venom plasma was examined by both in vitro and in vivo assays. ELISA is based on the horse anti-Naja antibodies recognizing toxic fractions mixture, the main antigen that induces neutralizing antibody against Naja naja oxiana venom. Consistent results were found between the ELISA optical densities and corresponding neutralization potency values, demonstrating that the in vitro assay as well as in vivo assay can be used to estimate neutralizing antibody activity of the sera. It was found that venom antibodies reached a maximum level about 3rd week after immunization. The major advantage of designed ELISA is its ability to correctly separate poor or non-responder horses during initial steps of immunization which venom antibodies are very low in sera and antivenom estimation is difficult by in vivo neutralization
    Keywords: Naja naja oxiana, Snake venom, Polyvalent antivenom, ELISA, potency
  • Seydeh Mahnaz Mousavi, Mehdi Zeinoddini *, AliReza Saeeidinia, Nasrin Xodadadi Pages 139-146

    Vibrio cholerae is one of the most important infectious human pathogens among the toxicogenic strains during the history of many pandemics and is a global threat to public health, especially in developing countries. Therefore, rapid and on-time detection of this infectious pathogen is necessary to prevent its outbreak. The aim of this work was the identification of zonula occludens toxin produced by V. cholerae using the LAMP technique.In this study, zot (zonula occludens toxin), one of the virulence factors of V. cholerae, was selected as the target gene and specific primers for the sequence of this gene were designed using the PrimerExplorer V5 software. The optimization of various factors affecting the LAMP reaction including Mg2+ ion, primers, temperature, and incubation time was performed in a traditional way and also by Taguchi test design. Finally, the LAMP products were visualized by agarose gel electrophoresis stained with ethidium bromide and SYBR Green I fluorescent dye. The data showed that the optimum condition for the LAMP reaction was 4-12 mM Mg2+ ion, 1.6-0.8 and 0.53 μM FIP/BIP, 0.4-0.2 and 0.13 μM F3/B3, temperature of 60-65 °C, and incubation time of 30-90 minutes. However, using Taguchi method, the optimum condition was 6 mM MgSO4, incubation time of 60 min, and temperature of 65 °C. In conclusion, the results of this study showed that the LAMP method provides the rapid, sensitive, and specific detection of zonula occludens toxin-producing V. cholerae and can be used for the design of an identification kit of this pathogen.

    Keywords: Zonula occludens toxin, Detection, Optimization, Taguchi
  • Mansoureh Ghorbani, Roohullah Hemmati *, Behnaz Saffar, Mojtaba Mortezavi Pages 147-154
    Phytase hydrolyzes phytic acid. Phytic acid is the main form of phosphorus storage in plant seeds in cereals, legumes, and oilseeds. In addition, phytic acid chelates essential minerals and binds to amino acids and proteins and prevents the action of digestive enzymes leads to decrease in the digestibility of proteins. Monogastric animals are unable to hydrolyze phytic acid due to the lack of phytase in their digestive system. In the current study, based on the docking results and DynaMut webserver predictions, T312R and F260R mutants, were chosen. Then, using Quick-Change PCR, the two residues in the active site of phytase from Yersinia intermedia phytase were mutated to Arg. Moreover, a double mutant F260R/ T312R was generated. Afterwards, the recombinant mutant and wild-type phytases were expressed in bacterial host, purified and their activities were measured. According to the results, the activities of T312R and F260R almost remained unchanged; however, that of T312R/ F260R was increased about 2.35 folds compared with the wild-type. This indicated that incorporation of two positively-charged arginine residues in the active site led to more interactions of these residues with the negatively charged-rich substrate and increased the phytase activity. In conclusion, highly active T312R/ F260R double mutant can be used as feed additive in feed industries.
    Keywords: Phytic acid, Phytase, Targeted mutagenesis, Quickchange, Kinetics, Catalytic efficiency
  • Seyedeh Sara Karimian*, MohammadTaghi Akbari, GholamReza Javadi, Seyed Saeed Sadr Pages 155-162

    Schizophrenia (SZ) is a chronic multi genetic and multifactorial neurodevelopment disorder. Neural cell adhesion molecule (NrCAM) gene, which was assessed in this investigation, has significant functions in the generation of neural cells. NrCAM interferes in synapse generation, axon guidance, proliferation, myelination and differentiation [1]. NrCAM mRNA differentially expresses while cortical neuron growth (GEO accession number GSE102350). Also, it has been proven that, malfunctions in the structure of neocortical cells associated with schizophrenia [2]. NrCAM involves in many signaling pathways and has been identified as an SZ biomarker in many studies [3, 4].


    This investigation intended to find significant changes in NrCAM protein expression in blood serum of SZ patients. Another purpose of this investigation is to evaluate the association of variant rs10235968 with NrCAM protein expression in patients' serum.


    In this study, 52 paranoid patients, and 33 healthy persons were selected. To evaluate protein expression, ELIZA method was used. variant data of our previous study were used in this study (5). All data were interpreted using the unpaired t-test, ANOVA, and Chi-Square test using GraphPad Prism8.4.3.


    NrCAM protein expression in blood serum has shown a significant difference between patients and healthy group (P = 0.0509). The association of variant rs10235968 and NrCAM protein expression has not detected (P = 0.9468). In our research, for the first time, NrCAM expression was assessed as an SZ candidate biomarker in blood serum of Iranian patients.

    Keywords: NrCAM protein, Schizophrenia, Biomarker, Blood serum
  • Fatemeh Ranjbaran, Ahmad Homaei*, Fatemeh Shayesteh Pages 163-172

    Due to the economic importance as well as the fact that microbial proteases are stable at the desired pH and tolerate harsh conditions, this study was conducted to isolate and identify bacteria that produce protease enzyme symbiosis in mangrove trees. In this study, protease-producing bacteria coexisting with the roots and leaves of mangrove trees were collected from the west coast of Qeshm Island (Konar Siah) and cultured them at 30 °C for 72 hours on nutrient agar medium.. Protease-producing bacteria were then screened by specific culture medium containing Skim milk agar and protease enzyme activity of the isolated bacteria was measured using 1% casein substrate. Finally, the bacterium with the highest enzymatic activity was selected and identified by 16SrRNA gene sequence analysis. The results of the analysis showed that 31 of the 46 bacteria isolated from mangrove leaves and roots were capable of producing protease enzymes. Among the six strong bacteria, one with the highest enzymatic activity was selected, which after molecular processes and sequencing led to the identification of the bacterium Kocuria sp. strain HR12. Kinetic studies revealed that the protease enzyme isolated from kocuria sp. HR12, exhibited its maximum activity at 50 ˚ C and pH 9.. The findings of this study provide a platform for the mass production of temperature-resistant protease enzymes from the coexisting bacteria of the Persian Gulf mangrove trees, which offers a wide range of potential uses for practical enzyme production.

    Keywords: Isolation, Identification, Protease producing bacteria, Mangroves, Optimal stability
  • Sara Kaveh, Elham Tazikeh Lemeski*, Elham Moniri, Homayon Ahmad Panahi, Davoud Beiknejad Pages 173-181

    Carboplatin is a well-known anticancer drug. It, as a targeted medicine, is loaded onto branched graphene oxide nanoparticles (GONPs). In the present study, polymeric coatings with chitosan (CS) bonded of the fifth dendrimer’s generation containing the groups of ethylenediamine (EDA) and methyl methacrylate (MMA) function as a responsive polymer. In order to formulate the dendrimer coating, the graphene oxide surface’s last carrier, the precursors are polymerized. Chitosan (polymer sensitive to pH) coating is then applied to the branched graphene oxide surface. Further, the drug release is raised. The final product’s average size is 42 nm. The FTIR, TGA, XRD, and SEM-EDX are applied to characterize its various adsorption properties. Physical factors of several adsorption processes, including time, pH, and temperature, are examined applying the final adsorbent. The drug release using the adsorbent coated with a polymer sensitive to pH is more than that without. Drug release tests show pH sensitivity behavior in graphene oxide/dendrimer/chitosan (GO/dendrimer/CS) with %98.8 drug release at acidic (pH = 5.6) and %90 at neutral (pH = 7.4) settings, i.e., in the tumorous cell and the blood pH levels, respectively.

    Keywords: Graphene oxide, Drug delivery, pH-Sensitive polymer, Dendrimer nanoparticles, Carboplatin, Chitosan
  • Arezu Hematian, Masoumeh Valipour*, Leila Hassani Pages 182-189

    In vitro protein aggregation is affected by many different factors such as the presence of crowder agents. As we know the intracellular environment is highly crowded and it contains high concentrations of macromolecules. Molecular crowding decreases the effective volume available for the proteins and affects protein–protein interactions like protein aggregation. Aggregation of proteins may lead to conformational changes and consequent conformational diseases. It is possible to mimic crowding condition in vitro by adding inert molecule such as polyethylene glycol (PEG). In this study, the effect of different concentrations of PEG was evaluated on β-lactoglobulin (BLG) aggregation at different pHs. It was also aimed to see if the environmental factor like pH could affect the protein aggregation. BLG aggregation was detected by UV-vis spectroscopy. The protein conformational changes were also examined by spectrofluorometer. SDS-PAGE method was applied to verify the disulfide bonds involvement in BLG aggregation. According to UV-vis spectroscopy data, BLG absorbance increased at higher concentrations of PEG. PEG induced aggregation was also influenced by physical parameter pH. Based on fluorescence results, PEG affected BLG compactness. SDS-PAGE showed that increasing of protein concentration induces more chemical aggregation. As a conclusion, crowding agent, PEG, induces protein aggregation and pH affects this process. This causes protein conformational destruction and may alter BLG function.

    Keywords: Macromolecular crowding, β-lactoglobulin, Aggregation, Polyethylene glycol