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Optimization of Aqueous Enzymatic Extraction of Flat-Europe Hybrid Hazelnut (Corylus heterophylla Fisch. × C. avellana L.) Oil and Analysis of Its Components
No full textAim: The research aimed to evaluate the technological factors of the composite enzymatic extraction of Flat-European hybrid hazelnut oil, and analyze the fatty acid composition, to lay the foundations for the industrialized production of this oil.
Study Design: Based on single factor tests, the best conditions for the aqueous enzymatic extraction of the oil determined by response surface analysis.
Place and Duration of Study: Food Science College, Shenyang Agricultural University between March 2014 and December 2015.
Methodology: The composition of Flat-European hybrid hazelnut was determined using GC-MS.
Results: Selection of suitable commercial enzymes is an important factor that can affect the extraction efficiency. Based on single factor tests, best conditions for the aqueous enzymatic extraction of the oil, determined by response surface analysis, were as follows: pH 6.13, enzymatic hydrolysis temperature of 59.9°C, dose of papain equal to 6% of seed weight, dose of α-amylase equal to 4% of seed weight, seed/water ratio of 1:5 (w/v), and enzyme hydrolysis time of 4.78 hours. By means of GC-MS analysis oleic acid (77.22%), linoleic acid (14.06%), linolenic acid (0.13%), palmitic acid (5.04%), stearic acid (2.35%), and arachidonic acid (0.44%) were identified as the major compounds of the flat-European hybrid hazelnut oil.
Conclusion: The mixture of papain (6% of seed weight) and α-amylase (4% of seed weight) was the best enzyme combination for the aqueous enzymatic extraction of flat-European hybrid hazelnut oil. The best technological parameters for the oil extraction process were: seed/water ratio of 1:5 (w/v), pH value of 6.13, reaction temperature of 59.90°C, and reaction time of 4.78 h. With these conditions, the oil yield was 82.36%, where unsaturated fatty acids were 92% of the total
Sensory Analysis of Porridges Processed from Flours of Palmyra New Shoots Enriched with Powders of Moringa oleifera Leaflets and Vigna unguiculata Beans
No full textThe current study focuses on the sensory properties of fifteen (15) composite porridges processed from flour of new shoots tubers of Borassus aethiopum Mart basis. The composite flours were processed from mixture of various ratios of B. aethiopum flour (BAM) and powders of beans of Vigna unguiculata (VUW) and leaflets of Moringa oleifera (MOL). Sensory analyses were performed to reveal the descriptive profile regarding four (04) sensory parameters, namely aroma, flavour, appearance, and texture, and then the general acceptance of the porridges. Thus, the sensory profile of the studied composite flour porridges was determined against two control flours (ET1 and ET2). Then, the general acceptance was probed with two of the composite flours highlighting better sensory profiles, and then compared to both control flours. From the sensory description of the studied porridges, the samples EF07 (57% BAM, 26% VUW, and 17% MOL) and EE09 (61% BAM, 24% VUW, and 15% MOL) provided higher sweet flavour (4.27 and 4.07/9), agreeable aroma (6.67 and 6.47/9), and fluid texture (6.27 and 5.53/9), and revealed lower bitter taste (1.73 and 1.80/9). Besides, composite flours porridges EF07 and EE09 were accepted by 86% and 82% persons including 46% and 24% full acceptance, respectively. Oppositely to the composite flours porridges containing no substantial ingredient, the control porridges (ET1 and ET2) recorded 100% acceptation by panelists. The formulations of composite flours with B. aethiopum Mart new shoots basis could be more accepted as food after addition of additives such as sugar, milk and natural aromas. Such processing could result in better promotion of this plant species culture for the biodiversity preservation and generating significant incomes for farmers in rural zones
Computational Molecular Analysis of the Sequences of PAPPA2 Gene of Selected Ruminants and Non Ruminants
No full textPregnancy associated plasma protein A2 (PAPPA2) is an Insulin-like growth factor binding protein (IGFBP) protease of the pappalysin family. This gene has been reported to be associated with prenatal growth, postnatal growth, skeletal growth, calving interval, milk yield, fertility and parity in cattle. The present study was undertaken to computationally investigate the attendant effects of the genetic variants of the PAPPA2 gene on its function and to gain insight into the evolutionary proximity and divergence in ruminants and non-ruminants at the studied locus. A total of fourteen (14) PAPPA2 nucleotide sequences comprising cattle (3), sheep (4), goat (1), pig (3), chicken (1) and horse (2) were retrieved from the GenBank. Functional analysis of non-synonymous single nucleotide polymorphism showed that eight amino acid substitutions (C31I, R60V, Y71G, S119L, S181Y, R190I, Q361M, P178_E180del) in goats, seven in sheep (V9E, L44D, T185N, A125W, Y78S, R194V, R240L), seven in cattle (L11F, C50_W51insTDAPK, E100L, A250T, G257L, M850V), nine in chickens (A60G, S104_R170del, A190F, G128A, I10C, S309G, F48L, Q1630L) and eight in pigs (A61L, P72D, L11Q, K184T, D110C, S193_R194insTQD, Q481_E484del and T170_V172delinsVA) were returned neutral suggesting their beneficial effect. The phylogenetic trees from nucleotide sequences revealed the close relatedness of members of the Bovidae family (sheep, cattle and goat). The present information may be exploited in research into the association between PAPPA2 genotypes and some important economical traits in farm animals
Investigation on the Effects of UV Radiation on Physiological Characteristics of Moringa oleifera Lam. in vitro and in situ
No full textMoringa oliefera seeds were treated with UV light type A, B and C for 30 minutes. Seedlings length, number of plants, number of axillary buds, number of adventitious buds and number of apical buds were recorded 34 days after sowing the treated seeds in addition to the control in vivo. Sterilized seeds were cultured in vitro on MS agar medium containing 4.0 mg/l BA for seed germination, then shoot explants were cut into small pieces and placed on the MS agar medium supplemented with 0.5 mg/l BA and 2.5 mg/l 2, 4-D. The percentage of callus induction and callus fresh weight were determined about six weeks after inoculating. Proline and carbohydrate concentrations were also determined for intact plant and callus cultures. Results showed that the physiological parameters studied in situ reduced significantly at UV-B recording 1.5, 1.8, 3.4, 2.8 and 19.3 cm No. germinated plants, No. Adventitious buds, No. axillary buds, No. apical buds and seedlings height respectively. UV-B recorded the highest mean values in relation to percentage of callus induction, callus fresh weight and proline concentration (100%, 112 mg and 9.7 µM/g respectively) compared to the control (72.3%, 93.3 mg and 7.3 µM/g respectively). A significant reduction in the mean carbohydrate concentration was observed in all UV treatments in both intact plants and callus cultures compared with control
Proteolytic Activity of Bacillus amyloliquefaciens UEF01 Endophytic to Carnivorous Plant Utricularia exoleta R. Br.
No full textAims: The endosphere of the carnivorous plant Utricularia exoleta R. Br. represents a unique niche for the study of microbial diversity. This study was aimed at to isolate and enumerate the protease producing potential of bacteria endophytic to U. exoleta R. Br.
Study Design: Extracellular proteolytic activity of the cell-free culture filtrate of the bacterial endophyte was determined following standard colorimetic assay using casein as the substrate.
Place and Duration of Study: The study was undertaken in the Microbiology Laboratory, Department of Botany, University of Calcutta during September 2015 and March 2016.
Methodology: A total of 36 phenotypically distinguishable bacteria endophytic to leaf, stem, bladder and fruit of U. exoleta were isolated and evaluated for protease production. The best protease producing isolate, UEF01 was selected, characterized and the conditions for protease activity were optimized.
Results: The selected isolate was characterized following morphological, physio-biochemical and 16S rRNA gene sequence analysis and identified as Bacillus amyloliquefaciens UEF01 (GenBank Accession No. KX816572). The crude protease of the cell-free culture filtrate of UEF01 was maximum at 35° C, pH 9 with 1.5% (w/v) casein. The enzyme appeared to be thermolabile with loss of >80% activity at 70°C (15 min incubation). Kinetic studies indicated the Km and Vmax values as 9.21 mg/ml and 71.43 U/mg of protein respectively. The enzyme was sensitive to Na and Mn ions as well as some selective protease inhibitors such as phenyl methyl sulfonyl fluoride, β-mercaptoethanol, and EDTA.
Conclusion: This proteolytic study will help in understanding the role of endophytes in digestion of prey within the bladders of the carnivorous plants.
 
Novel Method for Detection, Quantification and Cytopathic Changes in Developed Epithelial Cell Line with Human Papillomaviruses-16 Infection
No full textHuman papillomaviruses (HPVs) belong to the class of DNA viruses which are known to cause different human cancers. Development of new diagnostic methods and tools to study virus biology and mechanism of infection had been strongly felt in order to prevent HPV infections. In the present study we have designed new set of primers from E6 gene of HPV for its detection using PCR with an increased sensitivity of upto 0.2 pg. Hence, we established and characterized a new cervical epithelial cell line designated as CEC to be used as an in vitro tool for studying viral biology in order to prevent HPV infections. The interaction of HPV with CEC cells resulted in its neoplastic transformation and cytological changes such as multinucleation, nuclear irregularity, nuclear enlargement and perinuclear halos. CEC cell line was thus successfully established as an in vitro model system for studying interaction of HPV with squamous cervical epithelial cells
RNA-Seq Evaluating Several Custom Microarrays Background Correction and Gene Expression Data Normalization Systems
No full textMicroarray gene expression technologies represents a widely used tool in transcriptomics and genomics studies worldwide. Even if this technology exhibits a low dynamic range as well as a feeble sensitivity and specificity (limited performances) with respect to RNA sequencing (RNA-seq) methodology in whole transcriptomic and/or genomic studies; it is noteworthy to underline the stability of the former (microarrays) because of their well-established biostatistics and bioinformatics analysis schemes. Several studies shown that inadequate data pre-processing as regards microarray gene expression data analysis; i.e. inadequate gene expression data normalization (DN) and scarce noise background subtraction (BS), might compromise microarray aptitude in calling correctly significantly differentially expressed genes (DEGs). Here, we were interested in assessing the performance of 20 different microarrays background correction and gene expression data normalisation arrangements from R software “linear models for microarray and RNA-seq data analysis” package, by comparing the number of differentially expressed genes detected by our previous developed custom microarray designs and RNA-seq platform. The present study basing exclusively on several clustering and principal component analysis (PCA) as well as descriptive and inferential statistic surveys, developed in the R programing environment, suggested a predominance of microarray data normalisation systems with respect to noise background correction procedure. Although, all processed background subtraction and gene expression data normalization arrangement (BS+DN) claimed to improve the agreement (sensitivity) between microarrays and RNA-seq in calling DEGs; quantile normalisation procedure applied to our processed custom microarray designs has been recorded as exhibiting the best sensitivity (p-value<0.05), since discriminates the highest number of DEGs in agreement with RNA-seq as opposed to the others analysed microarray gene expression data normalisation systems. In conclusion our findings confirmed the pre-eminence of data pre-processing procedure in microarray gene expression profiling analysis according a priority to data normalisation procedure and suggested the stability of quantile normalisation system with respect to the others processed normalisation arrangements in the present executed gene expression comparative study. 
Molecular Differentiation of Five Quinoa (Chenopodium quinoa Willd.) Genotypes Using Inter-simple Sequence Repeat (ISSR) Markers
No full textKnowledge of genetic diversity is one of the important tools used for genetic management of quinoa accessions for plant breeding. This research aimed to molecularly characterize five quinoa genotypes using ISSR markers to reveal genetic polymorphism and identify unique markers for each genotype. Analysis of inter-simple sequence repeats (ISSR) revealed that 10 ISSR primers produced 53 amplicons, out of them 33 were polymorphic and the average percentage of polymorphism was 61.83%. The number of amplicons per primer ranged from 3 (HB-13, HB-10, HB-8 and 17898A) to 10 (HB-15) with an average of 5.3 fragments/primer across the different quinoa genotypes. Data showed a total number of unique ISSR markers of 24; eleven of them were positive and 13 were negative. Using ISSR analysis, we were able to identify some unique bands associated with quinoa genotypes. The genetic similarity ranged from 49% (between Ollague and each of QL-3 and Chipaya) to 76% (between CICA-17 and CO-407). The results indicated that all the five quinoa genotypes differ from each other at the DNA level where the average of genetic similarity (GS) between them was about 59%. The dendrogram separated the quinoa genotypes into two clusters; the first cluster included two genotypes (QL-3 and Chipaya). The second cluster was divided into two groups; the first group included two genotypes (CICA-17 and CO-407) and the second group included only one genotype (Ollague)
Molecular Markers and Postulation Study of Leaf Rust Resistance Genes in Various Egyptian Wheat Cultivars
No full textThe current investigation was carried out at seedling stage under greenhouse condition to: (1) postulate leaf rust resistance genes in ten Egyptian spring wheat cultivars at seedling stage and (2) Verify the presence of resistance genes in the selected cultivars using specific molecular markers linked to leaf rust resistance genes. The ten Egyptian spring wheat cultivars i.e. Misr 1, Misr 2, Sids 12, Sids 13, Gemmeiza 9, Gemmeiza 10, Gemmeiza 11, Sakha 93, Sakha 94 and Giza 168 as well as eight monogenic lines carrying single gene for leaf rust resistance i.e. Lr 21, Lr 25, Lr 32, Lr 35, Lr 37, Lr 39, Lr 47, and Lr 51 were tested at seedling stage using 15 leaf rust pathotypes i.e. BDGGK, CPPTB, DMKRT, FKRNK, FTFNB, KTSQG, LDDLD, LFGLL, NRRLS, NTTSR, PTKLN, STRTQ, TJKPR, TQTMJ and TTJBC. Eight specific molecular markers were known to be linked with the Lr,s resistance genes, i.e. Lr 21, Lr 25, Lr 32, Lr 35, Lr 37, Lr 39, Lr 47, and Lr 51 were used to verify the presence of these genes in the tested Egyptian wheat cultivars. The postulation test theorized the presence of Lr 25, Lr 32, Lr 35, Lr 37 and Lr 47 in Sids 12. Moreover, the wheat cultivar Misr 1 may have four genes i.e. Lr 25, Lr 35, Lr 39 and Lr 51. The wheat cultivars Misr 2, Sids 13, Sakha 94 and Giza 168 may have two genes i.e. Lr 35 and Lr 39 in the wheat cultivar Misr 2 and the same two genes Lr 35 and Lr 37 in the three wheat cultivars Sids 13, Sakha 94 and Giza 168. On the other hand, the wheat cultivars Gemmeiza 9 and Sakha 93 may have only one gene i.e. Lr 35 and Lr 32, respectively. While, the wheat cultivars Gemmeiza 10 and Gemmeiza 11 did not have any of the tested Lr genes under study using the fifteen tested pathotypes of P. triticina but it may be carrying some other genes. In the same way, all of the tested wheat genotypes may also carries additional resistant gene (s).The molecular markers survey with specific markers revealed that, the specific markers for Lr 21 and Lr 47 were detected in all tested cultivars. The marker for Lr 32 was recognized in four cultivars i.e. Gemmeiza10, Sakha 93 Sakha 94 and Giza168. The marker for Lr 39 was identified in all cultivars except Sakha 94. The markers for Lr 25, Lr 35, Lr 37 and Lr 51 were not found in any the tested cultivars. This study may add valuable information about resistance genes in the Egyptian wheat cultivars which may be used in pyramiding more genes in each cultivar or used as donor parents in different wheat genetic improving programs
Use of Gene Specific Universal Primers for Isolation of DNA Sequences Encoding Laccase Enzyme from a Wild Isolate of Schizophyllum commune
No full textLaccase enzymes plays a vital role in innumerable biotechnological applications and hence their large scale production has stimulated considerable research. In the present study, degenerate universal primer pairs were employed to isolate laccase gene from a wild isolate of laccase producing white rot fungi Schizophyllum commune. Primer pairs for this fungal isolate were also designed using laccase specific consensus sequences for fungi. The PCR product of 1000 bp amplicon was visualized on agarose gel using the degenerate primer pair Cu1F/Cu3R. Matching of the sequenced gel purified DNA sequence resembled most of the putative phosphatases involved in cell cycle with 100% identity to S. commune. Here we report the false negative results obtained upon use of laccase specific degenerate primers as well as other primers specific for the genus. These failed to contribute towards isolation of laccase gene even after optimization of PCR conditions in terms of reaction volume, annealing temperature, number of cycles, touchdown PCR and gradient PCR. These findings constitute a practical guide for researchers addressing amplification of transcripts of this biotechnologically important enzyme from the not so well characterized genus of Schizophyllum