Biotechnology Journal International
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Studies on the Proximate, Functional and Antioxidant Properties of Fermented and Unfermented Kariya (Hildergardia barterii) Seed Protein Hydrolysates Obtained by Enzymatic Hydrolysis
No full textThe degree of hydrolysis (DH), proximate composition, functional properties and anti-oxidative characteristics of enzymatic kariya seed hydrolysates were evaluated with a view to increasing the utilisation of the seed as food ingredient. Protein hydrolysates were obtained from fermented and unfermented kariya seed protein isolate using two proteolytic enzymes: Pepsin and pancreatin to obtain fermented pancreatin kariya protein hydrolysate (FPcKPH), unfermented pancreatin kariya protein hydrolysate (UPcKPH), fermented pepsin kariya protein hydrolysate (FPsKPH) and unfermented pepsin kariya protein hydrolysate (UPsKPH). The results showed that pancreatin hydrolysates had higher degree of hydrolysis (FPcKPH; 65.20% and UPcKPH; 50.50%) than pepsin hydrolysates (FPsKPH; 53.00% and UPsKPH; 33.93%) while the fermented hydrolysates showed higher DH than the unfermented hydrolysates both in the pancreatin and pepsin hydrolysates. The results also showed that fermented hydrolysates had better functional properties than the unfermented samples. The protein contents of the fermented hydrolysates were also improved. The antioxidant characteristics showed that fermented kariya protein hydrolysates exhibited higher DPPH (2,2-diphenyl-2-picrylhydrazyl hydrate) radical scavenging activity (FPcKPH, IC501.03; FPsKPH, IC501.40; UPcKPH, IC50 1.51; UPsKPH, IC50 6.97 mg extract/ml), metal chelating (FPcKPH, IC50 0.95; FPsKPH, IC500.53; UPcKPH, IC50 1.27; UPsKPH, IC50 1.06 mg extract/ml) and ferric reducing antioxidant power (FPcKPH, 0.63; FPsKPH,0.50; UPcKPH 0.35; UPsKPH, 0.23 AAµg/g). The study concluded that fermented kariya seeds hydrolysates could find applications as potential source of natural anti-oxidants in food
Sodium Alginate/Polyvinyl Alcohol Immobilization of Brevibacillus brevis OZF6 Isolated from Waste Water and Its Role in the Removal of Toxic Chromate
No full textAim: In this study we wanted to determine bacteria for chromium (VI) removal under pH, chromium concentration, carbon source and immobilizing agents.
Place and Duration of Study: This study was carried out in the Department of Biological Sciences, College of Natural and Applied Sciences, Crescent University, Abeokuta, Nigeria in the year 2015.
Methodology: Isolation of bacteria was done from industrial waste water of Abeokuta, Nigeria which is often released into water bodies and thus contaminates water during 2015. Nutrient agar added with 100 µg/ml Cr (VI) was used to isolate resistant bacterial strains. Resistance of the strains for Cr (VI) was evaluated on nutrient agar media. Natural material [sodium aliginate (SA)] and synthetic material (PVA)] immobilized bacterial cells for Cr (VI) removal experiment was done by 1, 5–diphenyl carbazide method.
Results: The strain OZF6 was characterized as Brevibacillus brevis using 16S rRNA gene sequence. All isolates (8 strains) were tolerant to chromium (VI). Among all strains, only Brevibacillus brevis OFZ6 reduced Chromium (VI). Brevibacillus brevis OZF6 reduced maximum Cr (VI) (72.5%) at pH 7. Brevibacillus brevis OFZ6 also reduced chromium (VI) significantly under various concentrations of chromium. Brevibacillus brevis OZF6 detoxified the metal 81% at 50 μg Cr/ ml, 75% at a concentration of 100 μg/ ml and 68% at 150 μg/ ml respectively. Among electron donors, maximum reduction was observed under influence of lactose. Among different matrices combinations for whole cell immobilization of OZF6, combination of 10% PVA, 10% sodium alginate (SA) proved to be best combination for Cr (VI) reduction.
Conclusion: Due to above properties, bacteria will be utilized for Cr (VI) detoxification in contaminated industrial waste water and thus will protect environment from contamination. There needs a proper regulation and treatment of these effluents prior their release into water bodies or into soil and thus ultimately protect population from carcinogenesis and other ill hazards
Identification and Enzymatic Potential of Bacillus Species Isolated from Traditional Cassava Starters: Potential for Attiéké Production
No full textAims: Microbial enzymatic activities are important in cassava dough fermentation for attiéké production. The objective of this study was to describe the molecular identification and the amylolytic, pectinolytic and cellulolytic enzymes potential of four (4) Bacillus species involved in cassava mash fermentation for the preparation of attiéké.
Place and Duration of Study: Laboratory of Biotechnology, UFR Biosciences, University Félix Houphouet-Boigny (Côte d’Ivoire), between February 2016 and April 2016.
Methodology: Four Bacillus strains (Bas 04, Bas 13, Bas 58 and Bas 66) used for this study were isolated from the traditional cassava starter ‘’mangnan’’. By using the PCR amplification of ribosomal 16S genes, Bacillus strains were identified. After culture in nutrient agar, strains were suspended in a tryptone salt solution. When we obtain Bacillus cells absorbance (turbidity) of 1.00 at 600 nm, 100 µL of this suspension were used to inoculate 5 mL of liquid medium containing peptone (1%), meat extract (1%), NaCl (0.3%) and 1% of starch or Pectin or Carboxylmethyl cellulose for enzymes production. The enzymatic activities were studied at different temperatures ranged from 25°C to 50°C and pHs (4.5, 5.0 and 5.5).
Results: Bacillus thuringiensis was identified with 99% to 100% of similarity, referring to the NCBI database. The best amylase, cellulase and pectinase activities were obtained with Bas 66 (3.80 U/mg), Bas 13 (0.45 U/mg) and Bas 58 (1.09 U/mg) at different optimum temperatures (25, 40 and 50°C) and pHs (4.5, 5.0 and 5.5), respectively.
Conclusion: These enzyme activities are important for cassava dough fermentation, using these Bacillus strains, contributing to the softening of the mash thus improving texture and allowing the digestibility of attiéké
Detection and Analysis of the Random Mutagenesis Site(s) of Xylanase Gene from Mutants of Bacillus subtilis subsp. spizizenii ATCC 6633
No full textAims: A total of five mutant strains of Bacillus subtilis subsp. spizizenii ATCC 6633 designated as the MXB 1, MXB 2, MXB 3, MXB 4 and MXB 5 were developed using random mutagenesis of ethyl methane sulfonate (EMS) and acridine orange (AO) in our previous study. Based on our present investigation, we identified, verified and sequenced xylanase gene of mutant strains of B. subtilis ATCC 6633 as the potent bacterial xylanase producers under submerged fermentation. Furthermore, amino acid analysis and comparison between the xylanases of the mutants and other xylanolytic bacteria were also elucidated. Overall, this study would provide gene and protein molecular information correlating nucleotide and amino acid structure related to the increased xylanase production by random mutagenesis. In respect to the objectives of this study, we compared the endoxylanase sequence of wild type B. subtilis ATCC 6633 with its mutants in order to determine their possible site(s) of mutagenesis and to analyse amino acid xylanase sequence of the mutants of B. subtilis ATCC 6633.
Methodology: After the verification of xylanase production by all mutants of B. subtilis ATCC 6633 on the xylan agar using Congo-red staining in the previous study, xylanase gene of the mutants was amplified from the genomic DNA to detect the mutagenesis site(s) by synthesizing primers directed against the sequence of xylanase gene obtained from the wild type of Bacillus subtilis subsp. spizizenii ATCC 6633.
Results: The comparison of xylanase genes from different mutants of B. subtilis and the wild type revealed the site(s) of mutagenesis. Interestingly, the mutations of the mutants of B. subtilis ATCC 6633 in this study were significantly reflected at the 5’ end of the mutants xylanase genes. The open reading frames (ORF) of the mutant xylanase genes ranged from 644 bp to 684 bp with translated encoding protein between 214 and 228 amino acid residues were obtained. On the other hand, predicted molecular mass from 23.94 kDa to 25.40 kDa and theoretical pI which ranged from 8.63 to 9.16 were attained from all of the mutant strains in this study. Based on the characteristics obtained, the mutant xylanases were suggested to belong to Glycosyl Hydrolase (GH) Family 11 with 98% homology to endo-1,4-beta-xylanase of B. subtilis subsp. spizizenii of W23. In fact, conserved regions, signal peptide, a cleavage site between the Ala28 and Ala29 residues and four Tyr residues specific to GH Family 11 xylanase were also observed and detected in all mutants. Indeed, two conserved glutamate residues of E94 and E183 that directly involved in the enzyme catalytic mechanism were also detected in the amino acid sequences of the mutants. The analysis of the deduced amino acid sequences revealed that the mutations in the signal peptide regions fostered increased hydrophobic core of xylanase residue. We suggested that these changes would probably be responsible for the increased extracellular xylanase yield in the mutants of B. subtilis. On the other hand, all the mutants of B. subtilis ATCC 6633 exhibited the tendency to be thermostable based on the Val, Ser and Thr frequency ratios which were almost identical to those of thermophiles. Furthermore, the increase of Thr to Ser ratio and presence of Arg residue found in the mutant strain of MXB 5 would enhance the polar interactions and hence improve the secondary structure stabilization that was usually one of the determining factors in the thermophilic proteins.
Conclusion: In a nutshell, the properties of B. subtilis mutant xylanases particularly mutant MXB 5 revealed its relevant potential in the biotechnology applications in bio-bleaching, textile, paper and pulp industries that commonly require high temperature usage in xylanase applications
Evaluation of in vitro Antifungal Activity of Silver and Selenium Nanoparticles against Alternaria solani Caused Early Blight Disease on Potato
No full textAim: This study investigated the effect of silver and selenium nanoparticles on Alternaria solani, the pathogenic fungus causing early blight disease of potato.
Place and Duration of Study: Drug Radiation Research Department, National Centre for Radiation Research & Technology (NCRRT), Atomic Energy Authority, Cairo, Egypt, 2013.
Methodology: The fungus was isolated from infected potato leaves that showed brown circular spots as early blight disease symptoms. Silver nanoparticles (AgNPs) were prepared biologically using gamma irradiated Trichoderma viride cell free supernatant. Selenium nanoparticles (SeNPs) were prepared by glutathione method. The synthesized AgNPs were characterized by UV-Vis spectroscopy, Dynamic Light Scattering (DLS) and Transmission Electron Microscope (TEM). Kocide® fungicide was used as reference.
Results: The fungus isolated of leaf spot was identified both microscopically and genetically as Alternaria solani causing early blight disease of potato. AgNPs were spherical in shape with average size of 12.7 nm. Selenium nanoparticles were prepared by glutathione as reducing agents. Under laboratory conditions, 25 µg/ml concentration of silver nanoparticles completely inhibited A. solani as compared to Kocide®, fungicide that gave maximum inhibition at 600 µg/ml. The selenium nanoparticles completely inhibited the fungal growth at 800 µg/ml.
Conclusion: AgNPs completely inhibited the growth of A. solani at low concentrations. Silver nanoparticles might be suitable alternative to chemical fungicides. While, SeNPs can be used as antioxidant for enhancing plant immunity
Ethnobotanical, Nutritional, Phytochemical and Antimicrobial Studies of Garcinia xanthochymus Fruit Extracts
No full textAim: An attempt has been made to document the ethnobotanical values of the fruits of G. xanthochymus in and around the Similipal Biosphere Reserve (SBR) forest and evaluate the nutritional, phytochemical and antimicrobial potential of fruit extracts of G. xanthochymus.
Study Design: Ethnomedicinal collection were made, followed by nutritional evaluation of the fruits. Phytochemical analysis was carried out by qualitative method and antimicrobial activity was evaluated.
Place and Duration of Study: Samples were collected from Similipal Biosphere Reserve and its adjoining areas during 2013 to 2014. Laboratory work was done at Ravenshaw University, Cuttack.
Methodology: Ethnobotanical data collections were made mainly through semi-structured questionnaires focusing on local name(s), present knowledge on uses as food, degree of uses, medicinal and economic values. Nutritional and qualitative phytochemical analysis was done by standard methods. Antimicrobial activity was evaluated against two Gram-positive bacterial strains, three Gram-negative bacterial strains and one fungal strain by disc diffusion method.
Results: Ethnobotanical survey suggested that the plant is widely used as a food or food supplement by the tribal people as well as in treatment of various ailments like fever, stomach problems, skin diseases and sexual disorders. Nutritional evaluation showed that the fruit was rich in carbohydrate and starch. The qualitative phytochemical analysis revealed that the fruits are rich in different types of bioactive compounds like saponin, tannins, alkaloid, terpenoid and phenolic compounds. The pathogenic strains showed concentration dependent susceptibility to crude extracts.
Conclusion: The results support the traditional use of the plant parts for the treatment of bacterial and fungal infections. However, further work on this fruit will open avenues for discovery of new lead molecules which might be used in pharmaceutical industries
Production and Characterization of Polyhydroxyalkanoate (PHA) Using Mango Seed Kernel as an Alternative to Glucose
No full textAim: To explore the possibility of using Mango (Mangifera indica) seed kernel as an alternative to glucose in the production of biodegradable plastic (polyhydroxyalkanoate) using soil isolated Bacillus megaterium.
Study Design: Experimental design.
Place and Duration of Study: This research was carried out at Biotechnology and Genetic Engineering Advanced Laboratory, Sheda Science and Technology Complex and Biochemistry Department, Federal University of Technology Minna, Nigeria, between the periods of June 2014 to October 2015.
Methodology: Using fed-batch fermentation technology, polyhydroxyalkanoate (PHA) pellets were produced, extracted and purified via solvent extraction. Polyhydroxyalkanoate produced was quantified using the crotonic acid assay at 235 nm by UV-VIS spectrophotometry. Fourier transform infrared spectrometry (FTIR) was used to characterize the PHA.
Results: Mango seed kernel produced a significantly greater (p<0.05) dry cell weight (DCW) and polyhydroxyalkanoate (PHA) (10.60±0.90 g/L; 64.22±0.55%) compared to glucose (10.19±0.01 g/L; 54.40±5.60%). Optimum conditions for maximum production of polyhydroxyalkanoate (PHA) were pH 7.5; temperature 35°C; substrate concentration of 2 g/100 ml; Bacilli inoculums volume of 2 ml and an incubation period of 72 hours. Fourier transform infrared spectrometry (FTIR) of extracted PHA revealed C-O, C=O, C-H, O-H functional groups at respective wave bands of 1125.5, 1644.37, 2938.65 and 3399.65 cm-1 for extracted mango seed kernel PHA; and 1086.92, 1646, 2937 and 3403.51 cm-1 for extracted glucose PHA.
Conclusion: Mango (Mangifera indica) seed kernel produced higher dry cell weight and polyhydroxyalkanoate than glucose hence, could be a substitute to glucose in PHA production
In vitro Cytological Studies of Leaf Callus Cultures of Orthosiphon aristatus (Blume) Miq
No full textThe study was carried out to unravel the in vitro cytological behaviour of leaf callus cultures of Orthosiphon ariststus (Blume) Miq. The cultures were raised using leaf explants on Murashige and Skoog (MS) medium supplemented with different growth regulators like auxins (2,4-D, IAA, NAA) and cytokinin (BAP) alone and in different concentrations and combinations. Best callusing from the leaf explants was achieved in 2,4-D supplemented MS medium at 5 mg/l concentration. Cytogenetic analysis of the chromosomal variations was carried out for primary cultures as well as 3-month-old callus cultures. The chromosome number did not show variations and the cells studied were observed to be diploid (2n= 28) in nature. Among the numerical observations induced, polyploid cells were the most frequent. The frequency of bridges was observed among the other structural changes. The composition and concentration of the growth regulators was found to affect chromosomal instability with multinucleate, multinucleolate and embryogenic masses. These observations indicate that genetic variations may arise during cell cultures
Effect of Stocking Density on the Performance, Carcass Yield and Meat Composition of Broiler Chickens
No full textBroiler chickens require adequate feed intake and uninterrupted emission of heat for intensive growth. An eight-week feeding trial was conducted to investigate the effects of stocking density on performance, carcass yield and meat composition of broilers. A total of 240 one-day old Arbor acre broiler chicks were randomly allocated to three stocking densities: Lower stocking density (LSD) of 10 birds/m2; Recommended stocking density (RSD) of 12 birds/m2; and Higher stocking density (HSD) of 14 birds/m2 respectively, with 4 replicates each having 20 birds per replicate. Feed and water were supplied ad libitum. Final live weight(FLW); Feed intake(FI); Feed conversion ratio(FCR); Mortality(%); Dressed weight(%); Prima cuts(%); Abdominal fat(%) and meat composition were assessed. Data were subjected to descriptive statistics and ANOVA ∞=0.05. Birds on HSD had numerically higher FLW (2262.80 g), better FCR (2.08) that was similar to that of LSD (2.07) and significantly lower mortality (0.00%). The values of the prime cuts (thigh, drumstick and breast), abdominal fat and meat protein were similar between birds on LSD and HSD. Up to stocking density 14birds/m2 broiler chickens’ performance and carcass characteristics were not negatively affected
Characterization and Antimicrobial Susceptibility Patterns of Isolates from Ward Fomites
No full textAim: The study was conducted to determine antimicrobial susceptibility patterns among isolates from ward fomites at Kiwoko Hospital and to detect resistances in the form of Macrolide Lincosamide StreptograminB (MLSB), Methicillin Resistant Staphylococcus aureus (MRSA), Extended Spectrum β Lactamases (ESBLs), AmpC, and Multi Drug Resistant (MDR) pathogens.
Study Design: Laboratory based cross-sectional study.
Place and Duration of Study: The study was conducted in various wards and sections at Kiwoko Hospital, a rural setting in the central region of Uganda, between January and June 2015.
Methodology: We recruited 290 samples from the Surgical, Medical, Maternity and Pediatric wards as well as the Out Patient Department (OPD) at Kiwoko Hospital for the study. Samples were taken by swabbing the different surfaces and instruments which included; sphygmomanometers, stethoscopes, beds, nurses’ stations, staff/visitors’ chairs, door handles, patients’ crepe bandages, curtains, switches, and sink handles among others. Susceptibility testing was done using the disc diffusion methods by Kirby Bauer for phenotypic expression of MLSB resistances, MRSA, MSSA, ESBL, MDR and AmpC. Co-resistances exhibited by isolated ESBL producers were also phenotypically tested.
Results: Of the 290 surfaces and instruments swabbed, 57.59% (CI= 49.18 - 67.01) carried bacterial pathogens and by using standard surface agar plating methods, Staphylococcus aureus was the mostly isolated pathogen 43 (25.75%), followed by Klebsiella pneumoniae 35 (20.96%), Escherichia coli 31 (18.55%), Pseudomonas aeruginosa 20 (11.98%), Enterococcus faecalis 12 (7.19%), Staphylococcus epidermidis 10 (5.98%), Proteus mirabilis 9 (5.39%), Bacillus spp. 4 (2.40%), and Staphylococcus saprophyticus 3 (1.80%). Among enterobacteriaceae, 5 (6.67%, CI= 2.16 – 15.56) were identified as AmpC producers and 16 (21.33%, CI= 12.19 - 34.64) as ESBL producers out of which 4/16 (25.00%, CI = 6.81 – 64.01) showed ESBL co-resistance. Of the 43 Staphylococcus aureus isolates, 9.30% were MRSA (CI = 2.53 - 23.82) and 90.70% MSSA (CI = 64.49 - 100). In MLSB resistance patterns, 23.26% of the total S. aureus isolates were constitutive MLSB while 6.98% showed inducible MLSB as 27.91% exhibited an MS phenotype. Out of all the isolates recovered from fomites, 27/167 (16.17%, CI = 10.65 – 23.52) were identified as Multi Drug Resistant (MDR).
Conclusion: Hospital fomites harbored resistant pathogens that could well persist for a long period of time thereby predisposing patients to Hospital acquired infections. Therefore, routine screening of clinical samples for MLSB, ESBL, AmpC, MRSA and MDR could significantly monitor potential treatment failures in the management of resistant bacterial infections spread by pathogens on ward items and surfaces at Kiwoko Hospital, Uganda