Research Journal of Biotechnology

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Vol. 15(12) December 2020

Purification and characterization of an extracellular ferulic acid esterase from Bacillus safensis SS3006

Singh Surabhi, Nigam Vinod Kumar and Sachan Ashish

Page No: 1-10

Abstract: Dietary ferulic acid (FA), a significant antioxidant substance, is currently the subject of extensive research that exists mainly as feruloylated sugar ester. To release FA from food matrices, it is necessary to cleave ester cross-linking by ferulic acid esterase (FAE). In the present study, the extracellular FAE from a Bacillus safensis bacterium was isolated, purified and characterized. FAE from the culture filtrate of Bacillus safensisSS3006 was purified to homogeneity by anion exchange chromatography and gel filtration chromatography. The protein corresponded to molecular mass of 35 kDa.

The enzyme was optimally active at pH 6.0 and temperature of 45°C. The purified FAE was fully stable at pH 5.0-7.0 and temperature up to 45°C after 4 hour incubation. Kinetic studies determined the Km and Vmax values of the purified FAE as 18 mM-1 and 222.25 μmol min-1 respectively. Metal ions such as K+, Mg++, Mn+, Fe++, Fe+++ showed a positive effect to enhance the enzyme activity of FAE whereas SDS and Hg++reduced the FAE activity.

Molecular characterization and genetic diversity assessment of ten plant species under family Combretaceae using ISSR markers

Das Arundhati, Bastia Akshayakumar and Rout Gyana Ranjan

Page No: 11-18

Abstract:The family Combretaceae is widely distributed in the tropical Africa, South America and Asia. This family having 20 genera and 600 species comprising herbs, shrubs trees and creepers distributed in tropical and subtropical regions, especially in Africa and very often in savannas. Both the genus Terminalia and Combretatum are the important among 20 genera. The genus Terminalia is considered as complex group having problems associated with taxonomic identification.

The present investigation is to assess the molecular diversity of genus Terminalia and Combretatum species based on Inter Simple Sequence Repeats (ISSR) markers. Seven Terminalia and three Combretatum species were analysed with 27 ISSR markers and to evaluate genetic relationships among and between the species. Out of total 27 primers screened, 10 primers were amplified across all species of which were polymorphic. Out of 101 alleles generated, 97 alleles were polymorphic and 04 alleles monomorphic among seven species of genus Terminalia and three species of genus Combretatum. Primer UBC-825 showed the highest polymorphism among 10 species for identification and diversity of plant species followed by UBC-827, UBC-864 and AM6 primers. On the basis of cluster analysis, it was observed that V1 (Terminalia arjuna) and V5 (Terminalia chebulla) closely resemble with other genotypes. The maximum similarity was obtained between V1 (Terminalia arjuna) and V5 (Terminalia chebulla) species. This study will help for identification of species as a diagnostic marker and conservation management strategies of forest species.

In vitro anti-diabetic and anti-inflammatory activities of metabolites isolated from Marine Sponge, Heteronema erecta (Keller, 1889) and its in silico studies

Ramachandiran Sivaramakrishnan, Sreechandan Rath, Kalimuthu Kapilan, Naidu Kavitha, Shankar Kanchana and Muthuvel Arumugam

Page No: 19-27

Abstract:Phenylalanine, aromatic amino acid and clupadonic acid, a long-chain fatty acid were purified from methanol and methanol-dichloromethane extracts of marine sponge H. erecta Keller. It was characterized by using UHPLC- Triple quadrupole ESI/MS. The clupadonic acid isolated possessed good inhibition towards α-amylase and α-glucosidase with IC50 of 17μg/ml and 22μg/ml respectively. Apart from that, phenylalanine gave better anti-inflammatory compared to clupadonic acid with 16μg/ml. The interaction between phenylalanine and clupadonic acid against the three receptors IXU7, 1SMD and 4Y14 has been studied using molecular docking.

Isolating anti Pseudomonas aeruginosa phages from wastewater and sewage samples: characterization of vB_PaeM_kmuV and vB_PaeM_kmuB

Amirpour-Rostami Sahar, Moeinzadeh Mahtab, Sadoughizad-Asl-Ajabshir Sona, Satarzadeh Naghmeh and Sabouri Salehe

Page No: 28-34

Abstract: In this study, 20 samples were searched for phages specific to Pseudomonas aeruginosa and 11 phages were isolated. Two phages were more effective on clinical antibiotic-resistant isolates. Therefore, further investigations were performed to characterize them. The electron micrographs showed that both phages belong to Myoviridae family with large heads and long tails. SDS-PAGE of the structural proteins also showed that both are large phages. Their genomes were sensitive to digestion by restriction endonucleases (HindIII, EcoRI and AluI). Both phages could tolerate a pH range of 3 to 9.5 and temperatures from 4 to 50°C.

Phage vB_PaeM_kmuB was adsorbed after 2 minutes and vB_PaeM_kmuV after 8 minutes of exposure to the host. Both phages could prevent the growth of the antibiotic resistant biofilm producing P. aeruginosa strain in liquid culture even at low multiplicity of infections. However, kmuB showed higher efficacy in this test. Phages kmuV and kmuB may be suitable candidates for phage therapy.

Biosorption of Lead from Aqueous Solution using a New Fungal Strain, Cunninghamella elegans TUFC20022

Jadhav S.K. and Mahish Pramod Kumar

Page No: 35-42

Abstract:The present work reports biosorption of lead metal ion using the biomass of Cunninghamella elegans TUFC 20022. Morphological and molecular characteristics of the fungus were studied. C. elegans TUFC 20022 was found to be tolerant against high concentration of lead metal ion and its live biomass absorbed the lead dissolved in the growth medium. Tolerance index (Ti) 92.3 was observed in 50 mgL-1 concentration of lead nitrate. Live biomasses absorbed 80.72% of lead from aqueous solution at pH 6 and 26°C incubation temperature. Maximum 90.34% lead from aqueous solution was absorbed by physically treated biomass while least absorption was obtained in detergent treated biomass (78.21%).

This biosorption is also tested with Langmuir and Freundlich models. Langmuir adsorption isotherm was found suitable with R2 of 98.9. Langmuir and Freundlich models also confirm favorable interaction and absorption by the fungal biomass. It is now concluded that the C. elegans TUFC 20022 is potential to remove lead from aqueous solution, so a technology based on this fungus will helpful to cleanup lead polluted water samples.

Modulation of ERF and ethylene biosynthetic genes during fruit development and jelly seed in mango

Ahmad Israr, Muthukumar M., Maurya P.K. and Rajan S.

Page No: 43-48

Abstract:Dashehari cultivar of mango is one of the leading mango varieties of North India and area under this variety is increasing in other parts of the country also. During fruit ripening, jelly seed formation has been experienced as one of the major disorders where Dashehari is commercially grown. However molecular mechanism behind this disorder is uncertain. To decipher role of ethylene forming enzyme isoform and its associated transcription factors in this disorder, we have mined 17 CDS of AP2/ERF genes in transcriptome data of mango jelly seed tissue. Expression analysis (RT-PCR) of four different ACS (ACS1, 2, 10, 12) and two different ACO (ACO1, ACO4), ERF21, ERF39 and IAA/AUX genes (AUX1, 2) was also analyzed during fruit development and ripening stages i.e. 0 dpa, 10 dpa, 20 dpa, unripe pulp, ripe pulp and jelly seed.

Expression of ERF21and ERF39 was found to be up-regulated during fruit development, ripening and jelly seed pulp as compared to 0 dpa. ERF21 and ERF39 plays important role during normal fruit development as well as mango ripening and jelly seed formation. ACS2 plays very important role during mango fruit development only while ACS10 and ACS12 are involved in development and ripening. AUX1 plays very important role during mango fruit development and ripening, however, AUX2 does not play any role in normal fruit development and ripening.

Prevalence and confirmation of brood diseases in apiaries of Himachal Pradesh, India

Thakur Meena, Negi Nitika, Negi Neha, Sagar Vinay, Sharma H.K. and Rana Kiran

Page No: 49-54

Abstract:The apiaries of Himachal Pradesh, India were surveyed for infestation of brood diseases of hive bees. For confirmation the disease pathogens were isolated and determined using RT-PCR assays for sac brood virus and morphological and PCR studies for European foul brood (Melisococcus plutonius). The primer pair SB1f/SB2r produced the clear and distinct bands of molecular sizes approximately 460 bp, 269 bp, 592 bp for district Solan, Kangra and Nauni, respectively from SBV infected samples of Apis mellifera. PCR studies with universal primer (BCF1 /BCR2) indicated the presence of Enterococcus faecalis in Apis cerana. In Apis mellifera maximum incidence of sac brood disease (4.33%) was observed in the month of June and minimum (0.50%) in the month of October, whereas no incidence was recorded in the month of November and December 2017 when rainfall (2.40-19.40mm) and temperature (12.53 to 14.12°C) were low.

The incidence of European foulbrood disease in A. cerana was maximum in the month of July, 2016 (23.00%) and minimum in the month of March, 2017 (0.44%) whereas, no incidence was observed in the month of December, 2016.

Identification of lncRNAs from Microarray Data for Schizophrenia: An In-silico Study

Neema Tufchi, Devvret, Pant Kumud and Waheed Syed Mohsin

Page No: 55-62

Abstract: Recent advances in various sequencing techniques have led to increased research in the field of lncRNAs (Long non-coding RNAs) with special context to diseases including schizophrenia. In the current study, GEO (Gene Expression omnibus) was explored and RNA sequencing data of lncRNAs was downloaded. The statistical analysis like univariate and multivariate Cox proportional hazards regression was performed on the data for the construction of survival based risk score.

For evaluating clinical and prognostic value of risk scores, Kaplan-Meier analysis has been done. Among 2,840 DELs (differentially expressed lncRNAs), two novel lncRNAs (NONHSAG044916.3 and NONHSAG004620.3) showed good prognostic effects in schizophrenic patients. Thus, this study helps in future exploration of prognostic lncRNAs for schizophrenia.

Isolation and presumptive identification of microbes in a typical fixed dome biodigester in Eastern Cape, South Africa

Mukumba Patrick, Makaka Golden and Tazvinga Henerica

Page No: 63-68

Abstract: A research was undertaken to establish the typical microbial composition in a field based biodigester as a possible avenue to optimize the degradation process. A 4m3 digester was fed with cow dung. Sampling for microbial and biochemical analysis was done at three different stages of the study period; day 0, day 21 and day 42. It was critical to identify native microflora prior to digestion at the peak and end of digestion based on ideal retention period. Temperature, biogas yield and pH readings of cow dung were measured to comprehend and establish their influence on biogas formation. Based on the microorganism’s morphology and biochemical properties, the microbes were identified from Bergey’s Manual of Determinative Bacteriology and Bergey’s Manual of Systematic Bacteriology.

The research findings were dominated by firmicutes with the remainder being proteobacteria. Eight different pure colonies were isolated and the microorganisms suspected to be in the biodigester were: Bacillus cereus, Bacillus thuringiensis, Escherichia coli, Paenibacillus alvei, Enterobacter cloacae, Clostridium perfringens, Enterococcus faecalis and Klebsiella oxytoca. It is envisaged that the outcome of this research will help in the formulation of a starter culture to speed up the digestion startup process and troubleshooting efforts requiring microbial intervention.

Interaction between Microarray Design Strategies and Data Pre-processing Procedures in Gene Expression Differential Analysis: A Computational Statistical Survey

Dago Dougba Noel, Dagnogo Olefongo, Loukou N’Goran Etienne, Koné Ali, Lallié Hermann Désiré, Mauro Giacomelli, Marinella Pinelli, Djaman Allico Joseph, Giovanni Malerba and Massimo Delledonne

Page No: 69-86

Abstract:The purpose of this study is to assess the link between microarrays design strategies and gene expression data pre-processing systems dynamisms. High throughput RNA-seq analysis is nowadays fully used by researchers in their transcriptomic as well as genomic studies. However, microarray remains a reliable and stable tool in transcriptomic and genomic surveys. It is well documented that microarray performance in transcriptomic and/or genomic studies is conditioned by oligonucleotide probe features (microarray design strategies) as well as by data pre-processing procedures, in term of background correction (BC) and expressed genes intensity data standardization (DS). After showing microarray data pre-processing dynamism, in gene expression differential surveys outcome, we embarked here in evaluating the interaction between microarray design strategies based on single and/or multiple short and/or long probe set, per gene model transcript and 20 different data pre-processing arrangements (BC+DS or DS+BC).

Findings show high performance in terms of results stability and sensitivity with regard to microarray design strategies based (i) on short multiple and long single probe set per gene transcript and (ii) long multiples oligonucleotides probes per gene model transcript respectively for background correction (BC) followed by expressed gene signal standardization (DS) (BC+DS) data pre-treatment procedure. Microarray data pre-treatment procedure based on gene expression data standardization followed by background correction (DS+BC), induced high variability in microarrays gene expression differential analysis outcome. R fitting curve analysis as well ANOVA test highlighted a good performances in terms of accuracy and sensitivity for microarray designs strategies based on short and long multiple oligonucleotide in gene expression differential analysis outcome respectively. In conclusion, presently processed microarray platforms exhibited stable attitude as well as high performance attitude, especially for microarray design strategies based on multiple oligonucleotide probe set per gene transcript model in differential analysis for the purpose of data pre-processed by standardization (BC+DS) as opposed to noise and/or background correction (DS+BC) procedures.

Finding potential inhibitors for HBB by molecular docking techniques involving toxicological study and its statistical significance

Phutela Simran, Sharma Archit and Mittal Amita

Page No: 87-98

Abstract:Sickle Cell Anemia (SCA), one of the first single gene disorder discovered, was found to be caused by β-globin mutation in Hemoglobin subunit beta protein that causes hemoglobin to form long rods that trigger red blood cells to turn into sickle-shaped. In-silico processes for drug discovery involve ligand preparation on the basis of Lipiski’s rule (of 5) and then docked with MVD (Molegro Virtual Docker) which was used during this research.

The best ligand is obtained on the criteria of MolDock score, Rerank score, Hbond score and subjected for their toxicological and drug likeliness studied through ADME when compared to already available FDA approved drugs and after their statistical significance was tested by performing T-Test. Ligand 2, Ligand 5 and Ligand 9 have shown better in-silico results in terms of MolDock score, Hbond score, toxicology and drug likeliness. T-test showed the reliable significance of these results. The above stated ligands can be taken for further investigation in the future to develop a more effective drug which can be a boon for sickle cell anemia suffering patients. In-silico drug designing lowers down the cost and comprises of less tedious steps than that involved in conventional drug discovery..

Removal of Dual and Multi-Species Biofilm using DNase I and Biocides

Sharma Komal, Singh Pagedar Ankita and Gupte Shilpa

Page No: 99-106

Abstract:Extracellular DNA is the most crucial component of the biofilm matrix found in various biofilm forming microbes. This extracellular DNA can be targeted to effectively remove the complex biofilms. The present study aims to evaluate the antibiofilm activity of DNase I against dual and mixed species biofilms and its treatment has increased their sensitivity for biocide killing. The preformed biofilms of microorganisms were first treated with DNase I (10µg/ml) and further treatment with biocides detached the biofilm effectively within 35-40 min. Among the dual, triad and quartet biofilms, the most effective results with DNase I were observed with peracetic acid (0.0157% (v/v)) which has given 92±0.9% biofilm removal of dual biofilm of S. aureus and S. typhi.

In case of triad biofilm of P.aeruginosa PAO1, E.fecalis and K.pneumoniae with pretreatment of DNase I, sodium hypochlorite (0.125 % (v/v)) was capable enough to detach biofilm 88±0.9%. Another notable biofilm removal (88±1.0%) was observed in quartet biofilm of P.aeruginosa PAO1, K.pneumoniae, E.faecalis and S.aureus using 1.4 % (v/v) Benzalkonium chloride whereas only 50% biofilm reduction was observed in non-sensitized biofilms. Therefore, observations revealed that DNase I exhibits potent antibiofilm and antimicrobial-sensitizing activities against mixed biofilms at clinically achievable concentration.

Micropropagation of Gerbera (Black jack) from in vitro culture using a standardized protocol

Mohapatra Manisha

Page No: 107-112

Abstract:Gerbera is a valuable ornamental species grown worldwide both through traditional and tissue culture techniques. The most efficient in vitro culture method commonly uses adventitious shoots, capitulum and leaf. In the present investigation, an attempt was made to develop an efficient method to regenerate Gerbera using petiole, leaf and capitulum as explants using various growth regulators. In case of capitulum, formation of callus occurs from receptacle portion in response to 6-Benzyl amino purine (BAP-0.4 mg/lit and 2.0 mg/lit) along with Naphthalene acetic acid (NAA-4.0 mg/lit) and Indol-3-acetic acid (IAA-2.0 mg/lit) respectively. The capitulum showed high rates of callus induction percentage in presence of 6-Benzyl amino purine (2.0 mg/lit) and Indol-3-acetic acid (2.0 mg/lit.).

It was observed that the capitulum and leaf explants produce more percentage of calli. Shoot organogenesis was observed on MS media supplemented with Kinetin (Kn-1.0-4.0 mg/lit.) and Indol-3-acetic acid (0.1-0.5 mg/lit.). High percentage of shoot formation was 94% on MS media with 2.0 mg/lit Kinetin and 0.1 mg/lit Indol-3-acetic acid. In case of root induction steps amongst Naphthalene acetic acid and 2, 4-dichlorophenoxyacetic acid (2, 4-D), highest root formation (89%) occurs when explants were cultured on medium supplemented with 0.5 mg/lit of Naphthalene acetic acid.

Characterization and efficacy of indigenous Azotobacter sp. isolated from different habitats for their nitrogen fixing ability and growth of rice

Sethi Santosh Kumar

Page No: 113-119

Abstract:Azotobacter fixes nitrogen to plant through nitrogen fixation process by reducing nitrogen to ammonia. In this study, different Azotobacter sp. were isolated from rhizospheres of different habitats mainly vegetables cropping fields and rice fields of Southern India by soil-plate method. These strains were identified based on phenotypic, biochemical and 16S rDNA sequences analysis. All the isolates produced indole-3-acetic acid (IAA). In the presence of tryptophan, plant growth promoting activities in terms of phosphate utilization, HCN production and also siderophore production were studied.

Out of ten isolates, five isolates are having highest nitrogenase activity. All the isolates are exhibiting nitrogenase activity and plant growth promoting activities having a promising potential strain for developing as plant growth promoting rhizobacteria.

Impact of Bioengineered Copper Quantum Dots on Germination, Photosynthetic Pigment and Antioxidant Response in Chick-pea under Dark Stress Environment

Rani Humaira, Gupta Rahul, Ansari Mohammad Israil and Singh Akhilesh Kumar

Page No: 120-130

Abstract:Cycles of light/ dark play a vital role in physiology/growth of plants by triggering different metabolic responses. Plants not only sense the commencement of dark stress environment but also utilize it for coordinating growth phenomenon. Likewise, copper as a vital micronutrient is needed by plants for mitochondrial respiration, antioxidant activity and so on. Currently, copper nanoparticles are receiving huge consideration owing to their economical, easier availability and so on. The present study examined the impacts of bioengineered copper quantum dots (Cu-QDs) in the range of 100-900 ppm on the seed germination, photosynthetic pigment as well as antioxidant response of darks stress environment exposed chick-pea (Cicer arietinum L.).

The supplementation of 500-700 ppm Cu-QDs for 72 h under dark stress was significantly found to boost the germination of chick-pea seeds over control (that lacking Cu-QDs treatment). The photosynthetic pigments (chlorophyll a, chlorophyll b and carotenoid) of dark and 600-700 ppm Cu-QDs treated chick-pea seedlings were also enhanced substantially over control. However, under the dark stress environment with the supplementation of 600-700 ppm Cu-QDs, the chick-pea seedlings recorded minimum antioxidant responses of reduced glutathione, glutathione reductase and ascorbate peroxidase compared to control. These results indicate the role of Cu-QDs as scavenging agent for reactive oxygen species (ROS). Overall, this investigation could be exploited to boost chick-pea seeds efficiency towards germination/ crop production during low-light intensity/ dark stress environment.

Screening and characterization of bioactive compounds produced by the moderate halophile Halobacillus sp. JS6

Yaradoddi Jayachandra S. and Mudgulkar Sulochana B.

Page No: 131-136

Abstract:A moderate halophilic bacterium Halobacillus sp. strain JS6 was isolated from the Arable soil sample of the west coast of Karnataka, India. The isolate, cream in pigment, produces circular colonies, gram-positive, aerobic, ferments several carbohydrates and has motile, pleomorphic cells and spore-forming catalase found to be positive. Apart from this, the isolate is able to grow even at 20% NaCl, pH10 and temperature about 450C. Antibiogram reveals the strain found to be resistant to cefepime, ceftazidime, colistin, cefotaxime and neomycin. The isolate is susceptible to ampicillin, chloramphenicol and ciprofloxacin.

The combination of phenotypic, biochemical studies and genotypic analysis by 16S rRNA analysis indicated the bacterium belonged to the genera Halobacillus with 98.73% gene sequence similarity through BLASTn analysis placed as a novel taxon under the genera and it was named as Halobacillus sp. strain JS6 and deposited in NCBI database under accession number HQ704870. The evolutionary significance of the isolated strain identified using Mega 4 software. The genus Halobacillus is known for its versatile features and can be explored for various biotechnological applications.

Molecular Genetic Analysis of Cedrus deodara (Roxb.) G. Don Age Class Population using cpSSR Markers

Kumar Akhilesh, Barthwal Santan and Ginwal H.S.

Page No: 137-142

Abstract:Cedrus deodara is an economically important conifer species in India and considered an endangered in Himalaya region. This study was planned to assess the haplotypic genetic diversity among different age class population of C. deodara to understand effect of time interval on gene diversity. cpSSR markers were used to study. Out of 33 cpSSRs, thirteen were found polymorphic and were selected for genetic diversity study. Among 90 genotypes, 48 size variant alleles were found. The Nei’s gene diversity (h) was higher (0.66) in the old age class populations compared to middle and young age class populations of Kathiyan forest Chakrata. The mean value of among population differentiation (GST) and gene flow (Nm) was found 0.11 and 4.46. Analysis of Molecular Variance (AMOVA) showed that most of the variations were found within populations (96.09%) as compared to among populations (3.91%).

The genetic differentiation (FST) was 0.04. The genetic relationships among the populations based on a dendrogram showing the similarities between populations were established using the Unweighted Pair-Group method with Arithmetic Averages (UPGMA). The dendrogram based on Nei’s (1972) genetic identity discriminated all three populations and clustered into one major cluster.

Statistical optimization of rhamnolipid biosurfactant for cost-effective production using Box–Behnken Design

Haque Ekramul and Hassan Saqib

Page No: 143-155

Abstract:The bacteria producing biosurfactant were isolated and screened for biosurfactant production using various qualitative assays from crude oil contaminated marine water sample. A potent bacterial isolate was selected on the basis of oil displacement activity and emulsification activity. Based on the sequencing of 16S rRNA gene of the isolate, ENO14 was identified as Pseudomonas aeruginosa.

Furthermore, the biosurfactant produced was confirmed to be the rhamnolipid molecule, a type of glycolipid biosurfactant by CTAB agar plate assay. In addition, rhamnolipid production optimization by Pseudomonas aeruginosa ENO14 was studied by Box-Behnken Design (BBD), a statistical model of (Response Surface Methodology) RSM. The two efficient models developed through RSM were observed to be satisfactory for depicting biosurfactant production with regards to rhamnolipid production (R2 = 0.9733) and emulsification activity (R2 = 0.9996). The highest rhamnolipid production and maximum emulsification activity were 7.93 ± 0.1 g/L and 182.51 ± 0.2 U/ml respectively which were continuously obtained under the optimized conditions of peanut oil cake (4 %), corn oil (0.8%), corn steep liquor (1.4 %). Therefore, high rhamnolipid yield may lead to its utility in petrochemical and food industry.

Regulation of cytokines by Argemone mexicana L. in potentiating the immune response

Goel Anjana

Page No: 156-162

Abstract:Argemone mexicana L.is used in folk medicines to treat many diseases in India. The decoction of leaves is used for the treatment of malaria, ulcers and chronic skin diseases. Aqueous leaves extract was investigated to validate the effect of the plant on hyperimmune disorders like allergy, asthma, arthritis, rheumatic arthritis etc. by regulating the expression of cytokines. Wistar albino rats were fed with nontoxic dose, 250mg/kg bodyweight of A.mexicana extract for 20 days. Spleens were removed from control and A.mexicana treated animals. Splenocytes were cultured in the presence of 10µg/ml con-A for 48 hrs. IL-2, IFN-gamma and IL-10 cytokines were quantitated in culture supernatant by ELISA technique. There was a significant increase in IL-10 expression in A.mexicana fed animals as compared to control animals.

In contrast, IL-2 and IFN-γ were expressed in significantly low quantities when compared with control animals. It can be concluded from the study that the effect of A.mexicana which is used to cure the allergic and hypersensitive type of immunological disorders of the skin like psoriasis might be due to the stimulation of IL-10, an immune-suppressive cytokine, secreted by TH-2 subset of TH cells while the cytokines secreted from TH-1 subsets, IL-2 and IFN-γ, which increase the immunological responses, were down-regulated.

Optimization of Lipase Production using Pseudomonas aeruginosa by Response Surface Methodology

Srinivasa Chary P. and Prameela Devi Y.

Page No: 163-166

Abstract:Lipases are the enzymes that catalyze the hydrolysis of triacylglycerols to free fatty acids and glycerol. In the present study, bacterial species from wastewater was isolated and screened for the best lipolytic strain (for lipase production, oil mill effluent was used as substrate) by optimizing its environmental factors. Media optimization of the enzyme was performed by using RSM statistical method that resulted in selection of the media composition for high production of enzyme.

Interference of Bacillus thuringiensis strain-AB3 on forensic DNA analysis of biological samples by producing Alpha hemolysin toxin

Bagban Mohammed Azim and Jain Nayan

Page No: 167-173

Abstract:Alpha-toxin is cytolytic to a range of cell kinds comprising human lymphocytes, erythrocytes, monocytes, platelets and endothelial cells. In the study, a bacterial isolate AB3 was obtained from biological evidence of simulated crime scene. Out of the 41 different colonies producing alpha-hemolysin most potent strain, AB3 was analyzed by gram staining, morphological characterization and 16S rRNA molecular identification. Multiple sequence alignment, phylogenetic analysis and construction of the phylogenetic tree were carried out by using the MEGA software version 6. Neighbor joining (NJ) phylogenetic tree was generated. Extraction of toxin from toxin producing microorganism which was determined by the zone of hemolysis on blood agar media was done by ammonium sulfate and sodium azide method.

Purification was done and confirmation of toxin was done by SDS-PAGE and HPLC. Purified toxin was mixed with biological samples (Blood, Saliva and Semen) in an appropriate ratio and qualitative and quantitative DNA analysis as well as RFLP analysis shows positive effect of alpha hemolysin toxin. From the study, it can be concluded that hemolysin toxin and toxin-producing microorganisms give an adverse effect on forensic DNA analysis as well as RFLP analhysis from blood, saliva and semen samples.

Standardization of Methodology for Extracting Ligninolytic Enzymes from Solid State Fermentation

Usha K.Y., Patil Sharangouda J., Dileep K., Shanti B., Rajasekhar Reddy B. and Praveen K.

Page No: 174-181

Abstract:The recovery of ligninolytic enzymes from fermented wheat bran derived upon growth of Stereum ostrea under optimal conditions was carried out with 20 ml of six different solvents – acetate buffer, phosphate buffer, citrate buffer, lactate buffer, succinate buffer and distilled water. Of all the solvents tested, phosphate buffer appeared to be the appropriate buffer for optimum recovery of all the three enzymes. Optimization of different volumes of phosphate buffer reveals that 25ml of solvent causes the maximum recovery of all the enzymes. Duration of soaking of fermented bran for recovery of enzymes was optimized by incubating fermented bran with solvent for different intervals - 30, 60 and 120 minutes. Thirty minute duration was optimum for maximum recovery of laccase, MnP and LiP while high protein content was recovered after 2 hours duration.

Of the two leaching conditions – shaking and stationary considered in the study, shaking was more effective for recovery of enzymes and proteins. Experiments were further conducted to find out no. of washes required for complete recovery of enzymes from fermented bran. Results of these experiment indicated that the first two washes were sufficient to totally extract the enzyme and protein from the bran.

Evaluation of Anticancer Activity and Apoptosis inducing ability of Suaeda monoica on Breast Cancer Cell Line

Swathi Minumula Sudarshan, Sathvika Chintalapani and Mangamoori Lakshmi Narasu

Page No: 182-186

Abstract:Breast cancer is the most frequently diagnosed cancer in women but rarely found in men. It is an invasive tumor majorly formed in lobules or ducts of the breast. The present study investigates the anticancer activity of Suaeda monoica with different solvent extracts against estrogen receptor-positive MCF-7 using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The different solvents used for this study were ethanol, methanol, acetone and diethyl ether. The diethyl ether extract of the plant showed the highest anticancer activity with an IC50 value of 186.78 μg/ml in comparison with other solvent extracts.

The morphological changes were then observed under an inverted phase-contrast microscope. Fluorescence imaging using propidium iodide stain was done to confirm the changes associated with apoptosis. The study shows that the plant has appreciable anticancer activity against the MCF-7 cell line and further the bioactive compound responsible for the anticancer activity should be isolated and purified.

Gamma irradiation enhancing value added architectural modification and tolerance to biotic constrains in rice (Oryza sativa L.)

Kumar Pankaj, Prasad Bishun D., Sahni Sangita, Ghatak Abhijeet, Pal Awadhesh K and Jambhulkar Sanjay J.

Page No: 187-198

Abstract:Various attempts have been made to determine the most effective mutagenic treatment for the induction of desirable traits in rice. In present investigation, an attempt was made to find out the effect of gamma irradiation for architectural improvement as well as tolerance to biotic stresses in Rajendra Mahsuri-1, the mega rice variety of north east plain zone of India. Lethal dose (LD50) of irradiation was determined initially by estimating percent survival of germinated seed, which was found at 350Gy. Upon increase in gamma irradiation (350Gy to 500Gy), the M1 population showed reduction in field survival, variation in effective tiller production and plant height. In M2 population, early vigour and number of chlorophyll mutants (albino) were counted at seedling stage; however, plant height, panicle length, early maturity, grain quality and analyses for insect and pathogen were performed at different stages of crop.

Indication of value added architectural attributes using gamma irradiation was revealed with significant reduction in maturity duration, plant height, increase in panicle length, effective number of tillers, improved shape and size of grain and low insect and disease problem in different mutant lines. For early maturity category, the lower irradiation (350Gy) rendered significantly reduced disease occurrence (P < 0.01). Similar result was obtained for the M2 mutants producing except for bacterial leaf blight which was higher at lower irradiation. The information generated through this investigation could be useful in breeding programme for rice improvement.

Rapid in vitro seedling establishment of Polygonatum verticillatum (L.) All.: an astavarga medicinal herb

Tiwari Tanuja and Chaturvedi Preeti

Page No: 199-207

Abstract:The present investigation was made to find out the best suited treatments of plant growth regulators and storage conditions for enhanced in vitro seed germination of Polygonatum verticillatum (L.) All., a valuable medicinal herb of Asparagaceae family. P. verticillatum shows pronounced seed dormancy and takes more than two years to attain crop maturity after sowing under natural conditions. Cold moist stratification of P. verticillatum seeds at 4±10C for 4 weeks followed by gibberellic acid (100 mg/L) treatment for 15 days and then their germination in 100 mg/L gibberellic acid (GA3) and 1mg/L kinetin (Kn)+1mg/L indole-3-acetic acid (IAA) supplemented Murashige and Skoog (MS) medium under 16/8h light/dark condition resulted in early and uniform emergence of hypocotyls, epicotyls and coleoptiles leading to improved seed germination of this plant.

Cold moist stratified seeds (with GA3 pretreatment) germinated in 50 to 100 mg/L GA3 supplemented MS medium were found to show significantly enhanced mean germination rate and seed vigor index (p<0.05). GA3 at a level of 100mg/L was found effective for both pretreatment of seeds as well as a supplement in MS medium. Using the above mentioned treatments, time requirement for germination and seedling establishment of this valuable medicinal herb was reduced to less than half the time required in natural conditions.

Protoplast mediated initiatives and the status of bio-techniques for improvement of table-purpose crop varieties

Yogranjan, Satpute Gyanesh K. and Tiwari Sharad

Page No: 208-219

Abstract:Plant protoplasts present a characteristic single cell system to reinforce several aspects of current biotechnology. The elite use of protoplast culture and subsequent fusion puts forward an opportunity for circumventing barriers to sexual reproduction and allows for gene transfer of nuclear and cytoplasmic genomes to enrich the gene pool of targeted crop species. Moreover, protoplast fusion effectively generates novel germplasm for selected breeding of conventional crosses and promotes crop improvement in existing cultivars. Reliable procedures are available to isolate and culture protoplasts from a range of plants including fiber crops and vegetables. Several parameters particularly the source tissues, procedures of protoplast isolation, culture media, environmental factors and experimental systems influence the ability of protoplasts and protoplast-derived cells to express their totipotency and to develop into fertile plants.

To generate transplastomic plants, the transient and stable nuclear transformation of isolated DNA into protoplasts and also organelle transformation are routinely practiced. Isolated protoplasts are also exploited in numerous studies focused on membrane function, cell structure, synthesis of pharmaceutical products and toxicological assessments. The study documents the advances well as classical developments achieved during the last few decades in protoplast-based technologies applied in major vegetable species.

Chronic Obstructive Pulmonary Disease (COPD) Biomarkers and Treatments: A Comprehensive Quest

Dhamodharan Pavithra and Arumugam Mohanapriya

Page No: 220-232

Abstract:Chronic obstructive pulmonary disease (COPD) is a life-threatening lung disease which causes airway obstruction in humans. This inflammatory disease produces multiple inflammatory mediators at different stages of the disease and also plays a key role in COPD pathogenesis. Global Initiative for Chronic Obstructive Lung Disease (GOLD) classifies COPD at four different stages. For each stage, prescriptions will vary and the treatment relies upon the patient’s health condition. This review focuses on the targets and treatments available for COPD and investigates the antagonistic impacts of the medications utilized by COPD patients.

Also, this review covers areas such as etiological agents available for COPD, therapeutic targets accessible for COPD, emerging biomarkers for COPD, presently accessible medications and its adverse effects. Ayurvedic treatment regimens and some of the phytocompounds produced from natural sources exhibiting anti-oxidant and anti-inflammatory activities were also summarized. Ayurvedic treatments and phytocompounds from natural sources might also act as promising medication contender for COPD in future.