Research Journal of Biotechnology

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Research Journal of Biotechnology

Studies on effect of process parameters for enhanced production of novel glutaminase free L asparaginase from Erwinia aroideae NRRL B-136 using Taguchi’s experimental design

Goswami Rachna, Veeranki Venkata Dasu, Hegde Krishnamoorthy and Bamnia Meenakshi

Glutaminase-free L-asparaginase is an important therapeutic agent used for the treatment of various lymphoblastic leukemia diseases. Eight variables including chemical and physical parameters were examined for their significance on the production of novel glutaminase-free L-asparaginase from Erwinia aroideae NRRL B-136 by employing Taguchi experimental design at shake flask level. At an optimum circumstance of tryptone: 6 g L-1, yeast extract: 6 g L-1, KH2PO4:1.25 g L-1, L-asparagine:1.0 g L-1, inoculum size: 1% (v/v), temperature:28°C, agitation: 200 rpm and initial pH of 7.0, the maximum production of L-asparaginase was found to be 62.44 U mg-1 of protein. The production of L-asparaginase was increased by 7.9 fold as compared to the unoptimized production conditions (8.85 Umg-1). Production of L-asparaginase was performed in a batch bioreactor under optimal levels of parameters.

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Performance studies of polyethersulfone ultrafiltration membrane for fruit juice concentration and purification

GajendranT. and Stalin N.

Cellulose acetate (CA) membranes have been prepared by varying from 15 to 20 wt% with increment of 1 wt% using N,N– dimethylformamide as the solvent. The 2.5 wt% of pore forming agent polyethylene glycol 600 (PEG 600) was also added. Prepared membranes have been subjected to ultrafiltration (UF) characterizations such as compaction, pure water flux, water content, membrane hydraulic resistance and morphological studies. The porosity and pore size of the membranes also increased on the addition of PEG. The effects of CA content and additive PEG in casting solution and pressure on concentration of clarified juice are studied. The 15 wt% of CA and 2.5 wt% of PEG in same CA achieved high flux and higher retention for dextran solution.

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Antifungal activity of Lactic Acid Bacteria isolated from human microbiota

Kiran Fadime

The objective of this investigation was to evaluate the effectiveness of some human orginated lactic acid bacteria strains to reduce the fungal growth in various food samples for the purpose of improving their quality and safety. Total fifteen strains were performed according to their inhibitory activity and nine strains of them, showing a broad inhibitory spectrum against fungal strains of industrial relevance were selected. Lactobacillus fermentum was found as the most promising strain and the strongest inhibition was observed with cell free supernatant obtained after 72 hr incubation period against Penicillum sp. and Fusarium oxysporum. Characterization of the antifungal metabolites demonstrated that the activity probably might be related with a combined effect of either organic acids or other pH-dependent proteinaceous antifungal compounds. All pediococci strains and Lactobacillus fermentum have also proved successful in controlling the fungal growth in commercial bread, tomato puree, apple and carrot samples.

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Correction of Ocular Artifacts in EEG signal using Empirical Mode Decomposition and Cross-correlation

Murthy Keshava G. N. and Khan Zaved Ahmed

Eye blinks cause changes to the electric fields around the eyes and consequently over the scalp. As a result, electroencephalographic recordings are often significantly distorted and their interpretation becomes problematic. The electrooculograph is a signal captured around the eye region viz. vertical electrooculograph and horizontal electrooculograph. Since the electrooculogaph is captured close to the eye than the electroencephalograph, it can be presumed that the eye blink effects are stronger in electrooculogaph than in electroencephalograph. In this paper we propose a new technique for artifact correction. Correlation between the electro-oculograph signal and electroencephalograph signal is exploited. In this method empirical mode decomposition is used to generate the intrinsic mode functions. The correlation between the electro-oculograph signal and the intrinsic mode functions is obtained. The intrinsic mode functions with high correlation and the intrinsic mode functions generated thereafter are eliminated. Rests of the intrinsic mode functions are recombined to form electro-encephalograph signal with mitigated effects of ocular artifacts.

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Development and evaluation of Bridge ELISA for the detection and quantification of antierythropoietin antibodies in human and rat serum

Prabhakar, Singh Poonam and Ganesan M.

Assays for the analysis of antierythropoietin antibodies (anti-EPO Abs) currently suffer from a high degree of non specificity and are cumbersome and time consuming to perform. They are therefore not well suited for the analysis of large numbers of human sera samples, a task that has become increasingly important due to an increase in the number of patients developing anti-EPO Abs. The objective of this study was to develop and validate a sensitive and specific ELISA for the determination of anti-EPO Abs that would suit these purposes. In this new double antigen bridging ELISA, anti-EPO Abs binds via one site to recombinant human erythropoietin (rhEPO) immobilized to microtiter plates (MTPs) and by a second site to rhEPO labelled with Horse radish peroxidase (HRP). A rabbit polyclonal anti-EPO Ab purified by immune adsorption is used as reference antibody preparation. The dynamic range of this ELISA was 5–1600 ng/ml per assay calibrated with the reference antibody preparation. The assay was specific for anti-EPO Abs and did not react with other immunoglobulins (Ig) present in rat or human serum. The lower limit of detection (LLD) of the assay was 5ng/ml and the lower limit of quantitation (LLQ) was 10ng/ml. Anti-EPO Abs could be detected in sera of pure red cell aplasia (PRCA) patients.

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A Conformational Study of GM1 headgroup with amyloid β (1-42) protein to identify the molecular specificity and interactions in Alzheimer’s disease

Devarajan Shine and Sharmila Jeya Sundara

Amyloid beta protein (Aβ) aggregation is the hallmark of Alzheimer’s disease. Recent accumulating evidence suggested that a unique complex, monosialo ganglioside (GM1)-bound Aβ possesses a high potential to facilitate this protein assembly. This interaction is pre-requisite for the initial recognition and further progression of beta amyloid aggregation in neuron cells. However there are no defined molecular interactions that lead to conformational changes between beta amyloid (Aβ1-42) protein and the GM1 headgroup. Moreover, the sialic acid moiety in headgroup plays a major role in molecular recognition. In order to study an atomic level conformational basis of this particular complex, we conducted an all atom molecular dynamics simulation of beta amyloid (Aβ1-42) peptide with monosialo ganglioside headgroup in solvent system. Our MD simulation revealed that upon binding of GM1 to the peptide leads to the alteration in the α-helix backbone especially at the C-terminal. The complex was stabilized by a network of hydrogen bonds. However, the amino acids His13, Leu17 and Phe20 play a major role in determining the interactions and stability with GM1. Thus, our conformational analysis helps to identify the structural alterations and interacting points on amyloid beta protein by GM1 headgroup.

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Gene Cloning, Expression and Optimization of Phytase Production in Pichia Pastoris

Meenakshisundaram S.

The phytase coding gene (PhyA) of size 1.3 kb isolated from Aspergillus ficuum NRRL 3135 and cloned in Pichia pastoris strain GS115 as recombinant pPICZαA-phytase was used for phytase production. The extracellular phytase activity after optimization with different co-substrates induction strategy in 4 L fermentor (NBS) increased to 13250 U/ml which is 509 fold increases compared to the wild type. This activity was achieved after inducing GS115/pPICZαA-phytase with methanol along with sorbitol as co-feeding in fed batch fermentation at 200C. The cloned phytase gene will have a potential to produce the thermostable enzyme which could be used in food industry as animals diets.

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Formulation of potential biological control products by using carriers embedded with microbial consortia

Thakkar Aarti and Saraf Meenu

This study was conducted with use of formulative preparation of the two selected best treatments of consortia from our previous study and their shelf life in two carrier materials up to six months and used to control diseases and also tested as biofertilizer in field trials. The bioformulation of P. aeruginosa (MBAA1) + T. citrinoviride (MBAAT) and B. amyloliquefaciens (MBAA3) + T. citrinoviride (MBAAT) was evaluated in combinations with carrier material gypsum and charcoal for their efficacy against diseases caused by phytopathogens M. phaseolina and S. Sclerotiorum in Glycine max L. M. phaseolina showed 85.32% disease incidence and it was greatly reduced in the plants treated with MBAA1+MBAAT + charcoal (10.85% disease incidence) followed by MBAA1+MBAAT+ gypsum (11.32% disease incidence) in pot trials. Where as in field condition T1 (MBAA1+MBAAT+ charcoal) proved best in all biometric parameters than other formulations and no disease was observed in field condition in any of the plants treated with formulated products.

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Estimation of fasting Blood glucose levels by invasive and indigenously developed noninvasive technology and its correlation with the glycated hemoglobin (HbA1c) biomarker in healthy and diabetic subjects

AChowdhury Koushik Md., Srivastava Anuj, Sharma Neeraj and Sharma Shiru

This research article represents the cross sectional study about the fasting glycemic index of normal and hyperglycemic subjects by invasive and indigenously developed noninvasive technology. Moreover, correlation between the fasting glycemic index and glycated hemoglobin (HbA1c) biomarker profile of both the normal and diabetic subjects was analyzed and discussed. The fasting glycemic index measurement was performed by the indigenous developed noninvasive method which utilizes both the amplitude modulated ultrasound and infrared techniques. The standing wave pattern of the ultrasound causes molecular vibration in the focused zone. The infrared light based technology had been utilized to detect those glucose sensitive vibrational patterns to predict the glycemic index of the normal and diabetic subjects. Invasive fasting glycemic index and profile of glycated hemoglobin (HbA1c) biomarker were determined by the established invasive methods. The result emphasizes that both the invasive, noninvasive fasting glycemic index values change as the glycemic index changes. Similarly, the glycated hemoglobin (HbA1c) biomarker profile of the normal subjects varies largely as compared to the diabetic subjects.

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Isolation of a unique Phenol degrading bacterial strain Escherichia coli moh 1 from effluent of an edible oil industry in Chennai, India

Fayidh M. A., Kalleary Sabina, Kasirajan Sudharsan and Muthusamy Sukumar

A novel environmental bacterial strain Escherichia coli moh1 was isolated and characterized from oil industry effluent in Chennai, Tamil Nadu, India. Primary growth experiments showed that this strain has the ability to grow in presence of up to 800 mg/l of phenol in NB medium at 37°C with the initial inoculum of ~104 CFU. The degradation rate of phenol (200 mg/l) was checked through monitoring growth at 24h using the HPLC system. The results showed considerable decrease in phenol concentration which was obvious from the decrease in peak intensity, a distinct change in the retention time and peak profile indicating complete degradation of phenol.16SrRNA sequence of the isolate was deposited to GenBank at NCBI and registered as a unique phenol degrading strain.

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Drug Target Proteins Prediction with Network Topological Indices

Wang Tengjiao and Xie Hongwei

The identification of drug target (DT) proteins can certainly be beneficial for new drug development. The properties derived from protein sequencing information have been well analyzed and exploited in predicting DT proteins. In this paper, we combined the protein sequencing properties and the PPI (protein-protein interaction) properties to accomplish the DT protein prediction. The high likely non-drug target candidates are collected as negative training data based on the PPI properties. Thereby, drug targets prediction can be solved as a traditional supervised classification problem. In this paper, 102 potential drug target proteins are predicted to help identify new drug targets. To guarantee the classifier be not overfitting, we tested it by 10 fold cross validation. The results show that the accuracy of our classifier is 82.01%.

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