Exogenous application of salicylic acid ameliorates salinity stress in barley (Hordeum vulgare L.).

Barley (Hordeum vulgare L.) is a significant cereal crop belonging to Poaceae that is essential for human food and animal feeding. The production of barley grains was around 142.37 million tons in 2017/2018. However, the growth of barley was influenced by salinity which was enhanced by applying a foliar spray of salicylic acid. The current study investigated to evaluated the potential effect of SA on the barley (Hordeum vulgare L.) plants under salinity stress and its possible effects on physiological, biochemical, and growth responses. The experiment was conducted at Postgraduate Research Station (PARS), University of Agriculture; Faisalabad to assess the influence of salicylic acid on barley (Hordeum vulgare L.) under highly saline conditions. The experiment was conducted in a Completely Randomized Design (CRD) with 3 replicates. In plastic pots containing 8 kg of properly cleaned sand, two different types of barley (Sultan and Jau-17) were planted. The plants were then watered with a half-strength solution of Hoagland's nutritional solution. After the establishment of seedlings, two salt treatments (0 mM and 120 mM NaCl) were applied in combining three levels of exogenously applied salicylic acid (SA) (0, 0.5, and 1 mg L-1). Data about morphological, physiological, and biochemical attributes was recorded using standard procedure after three weeks of treatment. The morpho-physiological fresh weight of the shoot and root (48%), the dry mass of the shoot and root (66%), the plant height (18%), the chlorophyll a (30%), the chlorophyll b (22%), and the carotenoids (22%), all showed significant decreases. Salinity also decreased yield parameters and the chl. ratio (both at 29% and 26% of the total chl. leaf area index). Compared to the control parameters, the following data was recorded under salt stress: spike length, number of spikes, number of spikelets, number of tillers, biological yield, and harvest index. Salicylic acid was used as a foliar spray to lessen the effects of salinity stress, and 1 mg L-1 of salicylic acid proved more effective than 0.5 mg L-1. Both varieties show better growth by applying salicylic acid (0 mg L-1) as a control, showing normal growth. By increasing its level to (0.5 mg L-1), it shows better growth but maximized growth occurred at a higher level (1 mg L-1). Barley sultan (Hordeum vulgare L.) is the best variety as compared to Jau-17 performs more growth to mitigate salt stress (0mM and 120mM NaCl) by improving morpho-physiological parameters by enhancing plan height, Root and shoot fresh and dry weights, as well as root and shoot lengths, photosynthetic pigments, area of the leaves and their index, and yield attributes and reduce sodium ions.

Melatonin as a master regulatory hormone for genetic responses to biotic and abiotic stresses in model plant Arabidopsis thaliana: a comprehensive review.

Melatonin is a naturally occurring biologically active amine produced by plants, animals and microbes. This review explores the biosynthesis of melatonin in plants, with a particular focus on its diverse roles in Arabidopsis thaliana , a model species. Melatonin affects abiotic and biotic stress resistance in A. thaliana . Exogenous and endogenous melatonin is addressed in association with various conditions, including cold stress, high light stress, intense heat and infection with Botrytis cinerea or Pseudomonas , as well as in seed germination and lateral root formation. Furthermore, melatonin confers stress resistance in Arabidopsis by initiating the antioxidant system, remedying photosynthesis suppression, regulating transcription factors involved with stress resistance (CBF, DREB, ZAT, CAMTA, WRKY33, MYC2, TGA) and other stress-related hormones (abscisic acid, auxin, ethylene, jasmonic acid and salicylic acid). This article additionally addresses other precursors, metabolic components, expression of genes (COR , CBF , SNAT , ASMT , PIN , PR1 , PDF1.2 and HSFA ) and proteins (JAZ, NPR1) associated with melatonin and reducing both biological and environmental stressors. Furthermore, the future perspective of melatonin rich agri-crops is explored to enhance plant tolerance to abiotic and biotic stresses, maximise crop productivity and enhance nutritional worth, which may help improve food security.

Cellular Prion Protein Role in Cancer Biology: Is It A Potential Therapeutic Target?

Cancers are worldwide health concerns, whether they are sporadic or hereditary. The fundamental mechanism that causes somatic or oncogenic mutations and ultimately aids cancer development is still unknown. However, mammalian cells with protein-only somatic inheritance may also contribute to cancerous malignancies. Emerging data from a recent study show that prion-like proteins and prions (PrPC) are crucial entities that have a functional role in developing neurological disorders and cancer. Furthermore, excessive PrPC expression profiling has also been detected in non-neuronal tissues, such as the lymphoid cells, kidney, GIT, lung, muscle, and mammary glands. PrPC expression is strongly linked with the proliferation and metastasis of pancreatic, prostate, colorectal, and breast malignancies. Similarly, experimental investigation presented that the PrPC expression, including the prion protein-coding gene (PRNP) and p53 ag are directly associated with tumorigenicity and metastasis (tumor suppressor gene). The ERK2 (extracellular signal-regulated kinase) pathway also confers a robust metastatic capability for PrPC-induced epithelial to mesenchymal transition. Additionally, prions could alter the epigenetic regulation of genes and overactive the mitogen-activated protein kinase (MAPK) signaling pathway, which promotes the development of cancer in humans. Protein overexpression or suppression caused by a prion and prion-like proteins has also been linked to oncogenesis and metastasis. Meanwhile, additional studies have discovered resistance to therapeutic targets, highlighting the significance of protein expression levels as potential diagnostic indicators and therapeutic targets.

Physiological, Biochemical, and Yield Responses of Linseed (Linum usitatissimum L.) in α-Tocopherol-Mediated Alleviation of Salinity Stress.

Exogenous application of antioxidants can be helpful for plants to resist salinity, which can be a potentially simple, economical, and culturally feasible approach, compared with introgression and genetic engineering. Foliar spraying of alpha-tocopherol (α-tocopherol) is an approach to improve plant growth under salinity stress. Alpha-tocopherol acts as an antioxidant preventing salinity-induced cellular oxidation. This study was designed to investigate the negative effects of salinity (0 and 120mM NaCl) on linseed (Linum usitatissimum L.) and their alleviation by foliar spraying of α-tocopherol (0, 100, and 200mg L-1). Seeds of varieties "Chandni and Roshni" were grown in sand-filled plastic pots, laid in a completely randomized design in a factorial arrangement, and each treatment was replicated three times. Salinity significantly affected linseed morphology and yield by reducing shoot and root dry weights, photosynthetic pigment (Chl. a, Chl. b, total Chl., and carotenoids) contents, mineral ion (Ca2+, K+) uptake, and 100-seed weight. Concomitantly, salinity increased Na+, proline, soluble protein, peroxidase, catalase, and superoxide dismutase activities in both varieties. Conversely, the growth and yield of linseed varieties were significantly restored by foliar spraying of α-tocopherol under saline conditions, improving shoot and root dry matter accumulation, photosynthetic pigment, mineral ion, proline, soluble protein contents, peroxidase, catalase, superoxide dismutase activities, and 100-seed weight. Moreover, foliar spray of α-tocopherol alleviated the effects of salinity stress by reducing the Na+ concentration and enhancing K+ and Ca2+ uptake. The Chandni variety performed better than the Roshni, for all growth and physiological parameters studied. Foliar spray of α-tocopherol (200mg L-1) alleviated salinity effects by improving the antioxidant potential of linseed varieties, which ultimately restored growth and yield. Therefore, the use of α-tocopherol may enhance the productivity of linseed and other crops under saline soils.

Arsenic concentrations in soil, water, and rice grains of rice-growing areas of Punjab, Pakistan: multivariate statistical analysis.

Arsenic (As), a class-A human carcinogen, is ubiquitously present in the earth's crust and soil and may enter the air, water, and surface environments through different natural and anthropogenic sources. In this experiment, soil, irrigation water, and rice grains were sampled from conventional rice-growing areas of Punjab, Pakistan. Soil samples were collected from 0 to 15 cm surface soil of rice growing fields, and rice grains were collected from the same field at crop maturity. Irrigation water samples were collected from the source used to irrigate the respective rice fields. Coordinates of sampling locations were noted using a global positioning system, and a locations map was made using ArcGIS. Soil samples were digested in a microwave digester using aqua regia, and plant samples were block digested using nitric acid. Arsenic concentration was determined using an inductively coupled plasma mass spectrometer coupled with an auto-sampler and integrated samples introduction system. The mean concentration of As in rice grains, soil, and water samples was found within the safe limit set by WHO except for a sample from Narowal (148.54 µg l-1) that exceeded the irrigation water standard limit, i.e., 100 µg l-1 for irrigation water. Principal component analysis was performed to reduce the multidimensional space of variables and samples. Through the calculations of estimated daily intake, it has been revealed that the As levels measured in this study would only contribute a small amount (less than 5%) of the total recommended daily intake allowance.

Evaluation and optimization of nanosuspensions of Chrysanthemum coronarium and Azadirachta indica using Response Surface Methodology for pest management.

The rapidly emerging field of nanotechnology is considered an important achievement in the agriculture sector to increase the pest mortality rate and improve the crop production. The present study evaluates the novel pesticidal and anti-microbial activities of Chrysanthemum coronarium and Azadirachta indica in the nano-suspensions form. The anti-solvent precipitation method was used to formulate nano-suspensions proposed by Response Surface Methodology (RSM). Physicochemical nature of plant extracts and nano-suspensions was characterized through analysis of Zeta-sizer, FT-IR, and HPLC. Characterization results revealed a minimum particle size of 121.1 and 170.1 nm for Chrysanthemum coronarium and Azadirachta indica, respectively. The pesticidal activity of nano-suspension was performed against red flour beetle (RFB) and lesser grain borer (LGB) pests, which showed the maximum mortality rate of 100% with 100% concentration of plant extracts and nano-suspensions of Chrysanthemum coronarium and Azadirachta indica against both insects. In comparison, the combination of these both plant extracts revealed the maximum 100% mortality with a 50% concentration of nano-suspensions (mixing ratio 1:1) after 72 h. The antibacterial activity showed the maximum zone inhibition of 9.96 ± 0.17 and 14.17 ± 0.50 mm against S.aureus and E. coli with nano-suspension of Chrysanthemum coronarium, and 12.09 ± 0.11 and 14.10 ± 0.49 mm with nano-suspension of Azadirachta indica, respectively. It is concluded that individual nano-suspensions showed better pesticidal as well as antimicrobial activities than combinations. However, the constructed nanosuspension can be applied to control the plant pests and diseases simultaneously.

An Ignored Contributing Factor of Vitamin-D Deficiency, despite the Strong Association with Breast Carcinoma among Women in Punjab, Pakistan.

Pakistani females are at elevated risk of breast cancer, hence there is a need to explore every possible potential contributing factor. The present study was performed to analyze the perception of women about their Vit-D levels in both urban and rural areas. Women included 154 clinically diagnosed breast cancer patients from different hospitals and 248 randomly selected females as control group were recruited from Punjab, Pakistan. Statistical analysis was done using SPSS to find the association of Vit-D deficiency with breast carcinoma in different age groups. Out of a total of 402 respondent, 51.5% were completely ignorant of their Vit-D level. Pearson's Chi- square test for those who had perception about Vit-D deficiency among the breast cancer patients and control group revealed asymptotic 2-sided significance of 0.004 while among different age groups, 41 to 50 years were most prone to deficiency with P = 0.003. Urban women were found to had 12% greater association of Vit-D deficiency as compared to women living in rural areas. It was concluded that Vit-D deficiency is a highly contributing factor for breast cancer so every female must be aware of the importance of Vit-D and should maintain a sufficient level of this crucially important vitamin.

Structural and Functional Annotation of Napin-Like Protein from Momordica charantia to Explore its Medicinal Importance.

Seed storage proteins not just provide essential nutritional ingredients for growth of seedlings but also have their potential role in defense mechanisms of plants. Napin is a seed storage protein and belongs to 2S albumin family. Napin and napin-like protein have many biological defensive activities including antifungal, antimicrobial, trypsin inhibitor, and also act as antagonist of calmodulin. Napin protein possesses various isoforms with different biological activities. In this study, the protein sequence of napin from Momordica charantia was retrieved from GenPept database for characterization. A complete annotation of napin including its physicochemical properties was done. Three dimensional (3D) modeling and interactions of napin-like protein with other proteins were also predicted using various bioinformatics tools. A phylogram of napin-like protein from M. charantia with its homologs was also reconstructed to reveal its evolutionary relationships with napins and other 2S albumin proteins from various plants. The study has revealed the structural characterization, biological interactions, and evolutionary background which will play crucial role in exploring the medicinal and biological potentials of napin-like protein from M. charantia as well as worth of napin and napin-like protein has been disclosed.

Therapeutic Attributes of Endocannabinoid System against Neuro-Inflammatory Autoimmune Disorders.

In humans, various sites like cannabinoid receptors (CBR) having a binding affinity with cannabinoids are distributed on the surface of different cell types, where endocannabinoids (ECs) and derivatives of fatty acid can bind. The binding of these substance(s) triggers the activation of specific receptors required for various physiological functions, including pain sensation, memory, and appetite. The ECs and CBR perform multiple functions via the cannabinoid receptor 1 (CB1); cannabinoid receptor 2 (CB2), having a key effect in restraining neurotransmitters and the arrangement of cytokines. The role of cannabinoids in the immune system is illustrated because of their immunosuppressive characteristics. These characteristics include inhibition of leucocyte proliferation, T cells apoptosis, and induction of macrophages along with reduced pro-inflammatory cytokines secretion. The review seeks to discuss the functional relationship between the endocannabinoid system (ECS) and anti-tumor characteristics of cannabinoids in various cancers. The therapeutic potential of cannabinoids for cancer-both in vivo and in vitro clinical trials-has also been highlighted and reported to be effective in mice models in arthritis for the inflammation reduction, neuropathic pain, positive effect in multiple sclerosis and type-1 diabetes mellitus, and found beneficial for treating in various cancers. In human models, such studies are limited; thereby, further research is indispensable in this field to get a conclusive outcome. Therefore, in autoimmune disorders, therapeutic cannabinoids can serve as promising immunosuppressive and anti-fibrotic agents.

Immunogenic profiling and designing of a novel vaccine from capsid proteins of FMDV serotype Asia-1 through reverse vaccinology.

Foot-and-mouth disease is one of the devastating transboundary animal diseases causing heavy losses to the livestock industry. Different vaccines based on the inactivated FMD virus are used against this disease, but lack of immunological memory and the need for high biocontainment are the major drawbacks of these vaccines. A novel vaccine comprising recombinant antigenic regions is effective, as they lack viruses for production. Considering the fact, capsid proteins vp4, vp2, vp3, and vp1 with 3C protease of FMDV serotype Asia-1 were analyzed through reverse vaccinology approaches in this study. The sequence and structural analysis of the proteins is carried out through various bioinformatic tools and the sequence analysis has figured out the acidic nature and thermal stability of the proteins, likewise, the phylogenetic analysis helped us to trace the FMDV isolates, elucidating that selected proteins belong to the strain (Group VII), which is currently circulating in Pakistan. Next, the B-cell and MHC Class-I epitopes are identified from the antigenic proteins by immunoinformatic tools. The highly conserved, antigenic, and non-allergenic epitopes are used to design the vaccine. Accordingly, the codon adaptation and in silico cloning of the corresponding genes is performed. Thus, the bacterial expression vector could be used for efficient expression and large-scale production of the vaccine.

Isolation and characterization of antihypertensive peptides from soy bean protein

Proteins and peptides are the most diverse biomolecules found in nature and make our interest due to their wide applications in food and pharmaceutical industry. Angiotensin Converting Enzyme (ACE) plays a major role in controlling blood pressure. The inhibition of ACE with peptides is a main target in the regulation of hypertension. The objective of the present study was to investigate the therapeutic potential of soy bean. This was accomplished by isolation of ACE inhibitory peptides using response surface methodology (RSM) and characterization of these bioactive peptides by mass spectrometry. 31 hydrolyzed fractions were isolated and evaluated for their ACE inhibition potential. Hydrolyzed fraction having highest ACE inhibitory activity was characterized by liquid chromatography-mass spectrometry (LC-MS) technique. RSM results showed maximum ACE inhibition potential (64%) by hydrolyzate was obtained at 45 ºC temperature, pH 8.0, E/S 0.2 in 2 hours hydrolysis time. Results of LC-MS analysis revealed Ser-Gly, Ser-Pro, Met-Ala, His-Ala, Lys-Pro, Phe-Thr, Met-Leu, Pro-Arg, Ala-Pro-Val, Pro-Ala-Leu, Val-Met-Gly, Pro-Leu-Val, Pro-Pro-Gln, His-Arg-Gly, Ser-Phe-Val-Leu, Ala-Val-His-Try, Arg-Thr-Val-Arg, His-His-Tyr-Leu-Val, Asp-Gly-Ala-Cys-Ser-Ala-Asn and MetVal-Thr-Gly-Pro-Gly-Cys-His bioactive peptides in hydrolyzed fraction of soy bean. Our data provide evidence that response surface methodology is a good approach for isolation of antihypertensive bioactive peptides with more potent activity as nutraceuticals or pharmaceuticals. Therefore soy bean can be use for industrial production of pharmaceutical grade natural medicines for handling high blood pressure.

Genome-Wide Association Mapping for Stripe Rust Resistance in Pakistani Spring Wheat Genotypes.

Stripe rust caused by the pathogen Puccinia striiformis f. sp. tritici (Pst) is a major threat for wheat, resulting in low yield and grain quality loss in many countries. Genetic resistance is a prevalent method to combat the disease. Mapping the resistant loci and their association with traits is highly exploited in this era. A panel of 465 Pakistani spring wheat genotypes were evaluated for their phenotypic response to stripe rust at the seedling and adult plant stages. A total of 765 single nucleotide polymorphism (SNP) markers were applied on 465 wheat genotypes to evaluate their stripe rust response against nine races during the seedling test and in three locations for the field test. Currently, twenty SNPs dispersed on twelve chromosomal regions (1A, 1B, 1D, 2A, 2B, 4A, 4B, 5B, 6A, 6B, 6D and 7B) have been identified that were associated with rust race-specific resistance at the seedling stage. Thirty SNPs dispersed on eighteen chromosomal regions (1A, 1B, 1D, 2A, 2B, 2D, 3A, 3B, 3D, 4B, 5A, 5B, 6A, 6B, 6D, 7A, 7B and 7D) are associated with adult plant resistance. SNP loci IWB3662 was linked with all three Pakistani races, and likewise IWA2344 and IWA4096 were found to be linked with three different USA races. The present research findings can be applied by wheat breeders to increase their resistant capability and yield potential of their cultivars, through marker-assisted selection.

Development of Whey Protein Concentrate-Pectin-Alginate Based Delivery System to Improve Survival of B. longum BL-05 in Simulated Gastrointestinal Conditions.

Bifidobacterium longum BL-05 encapsulated beads were developed by using whey protein concentrate (WPC) and pectin (PE) as encapsulating material through extrusion/ionic gelation technique with the objective to improve survival of probiotics in harsh gastrointestinal conditions. B. longum BL-05 was grown in MRS (de man rogosa and sharpe) broth, centrifuged and mixed with polymeric gel solution. Bead formulations E4 (2.5% WPC + 1.5% PE) and E5 (2% PE) showed the highest value for encapsulation efficiency, size, and textural properties (hardness, cohesiveness, springiness) due to increasing PE concentration. The survivability and viability of free and encapsulated B. longum BL-05 was assessed through their resistance to simulated gastric juice (SGJ), tolerance to bile salt, release profile in simulated intestinal fluid (SIF), and storage stability during 28 days at 4 °C. The microencapsulation provided protection to B. longum BL-05 and encapsulated cells were exhibited significant (p < 0.05) resistance to SGJ and SIF as compared to free cells. Bead formulations E3 (5.0% WPC + 1.0% PE) and E4 (2.5% WPC + 1.5% PE) exhibited more resistance to SGJ (at pH 2 for 2 h) and at 2% bile salt solution but comparatively slow release as compared to other bead formulations. Free cells lost their viability when stored at 4 °C after 28 days but microencapsulated cells demonstrated promising results during storage and viable cell count was > 107 CFU/g. This study revealed that extrusion using WPC and PE as encapsulating material could be considered as one of the novel technologies for protection and effective delivery of probiotics.

Enhanced Production Of Streptokinase By Chemical Mutagenesis Of Streptococcus agalactiae EBL-20

Abstract Streptokinase (SK) is an enzyme that is used for the treatment of cardiovascular diseases. The current study focused on the enhanced production of SK by inducing mutation in Streptococcus agalactiae EBL-20 and optimization of medium components and culture conditions for the maximum growth of mutant derived strain. S. agalactiae EBL-32 was selected as a potent mutant after exposure of S. agalactiae EBL-20 to EMS for 180 minutes. SK activity obtained from mutant derived strain was found to be 1.6 fold higher as compared to the activity achieved by wild strain. Nutritional requirements of the mutated strain were optimized by single factor analysis method suggesting glucose as the optimum carbon source; yeast extract and peptone as a suitable nitrogen sources and corn steep liquor (CSL) as an appropriate substrate for the maximum SK production. The culture conditions determined by response surface methodology (RSM) suggested that a temperature value of 37.5⁰C and pH 7 of the fermentation medium with 2.50 mL inoculum size for 36 hours of incubation was optimum for maximum yield of SK. Hence the optimization studies resulted into 1.92 fold increase in the yield of SK suggesting the new isolate suitable for commercial scale production of SK.

Enhanced production of streptokinase from Streptococcus agalactiae EBL-31 by response surface methodology.

Streptokinase (SK) is a fibrinolytic protein used for the treatment of cardiovascular disorders. In the present study, enhanced production of SK was achieved by determining the optimum fermentation conditions for the maximum growth of Streptococcus agalactiae EBL-31 using response surface methodology (RSM). Four process variables (pH, temperature, incubation time and inoculum size) with five levels were evaluated in 30 experimental runs. Central composite rotatable design (CCRD) was employed to predict the effect of independent variables on SK activity. The statistical evaluation by ANOVA showed that the model was fit as the effect of single factors, quadratic effects and most of the interactions among variables. The value ofR2 (0.9988) indicated the satisfactory interaction between the experimental and predicted responses. Furthermore, the model F value (902.67) and coefficient of variation (1.92) clearly showed that the model is significant (p =>0.0001). The functional activity of SK was determined by spectrophotometric analysis and maximum SK production was obtained at pH-7.0, temperature- 37.5oC, an incubation time of 36 hours and 2.5 mL inoculum size. Hence it was concluded that the optimization of culture conditions through RSM increases the production of SK by 2.01-fold. Production of SK by fermentation is an economical choice to be used for the treatment of cardiovascular diseases.

Identification and lead-in characterization of novel B3 metallo-ß-lactamases.

Metallo-ß-lactamases (MBLs) are zinc ion dependent enzymes that are responsible for the emergence and spread of ß-lactam resistance among bacterial pathogens. There are uncharacterized putative MBLs in the environment and their emergence is major interference in the generation of universal MBL inhibitors so it is important to identify and characterize novel MBLs. In this study two novel MBLs from Luteimonas sp. J29 and Pseudoxanthomonas mexicana were identified using B3 MBLs as query in BLAST database search. 3D models of putative MBLs generated by SWISS-MODEL server taking AIM-1 as a structural template were verified using web based structure assessment and validation programs. Multiple sequence alignment revealed that residues important for substrate binding were conserved and loop region residues (156-162 and 223-230) important for catalysis are variable in these novel MBLs. Homology models showed typical MBL α/ß/ß/α sandwich fold containing six α helices, twelve ß strands and metal interacting residues are conserved in similar way as with other B3 MBLs. We report promising putative B3 MBLs with some variations and substrate docking studies revealed that novel MBLs have attributes close to acquired B3 MBLs.

Kinetics and thermodynamics of ethanol production by Saccharomyces cerevisiae MLD10 using molasses.

In this study, we have used ultraviolet (UV) and γ-ray induction to get a catabolite repression resistant and thermotolerant mutant with enhanced ethanol production along with optimization of sugar concentration and temperature of fermentation. Classical mutagenesis in two consecutive cycles of UV- and γ-ray-induced mutations evolved one best catabolite-resistant and thermotolerant mutant Saccharomyces cerevisiae MLD10 which showed improved ethanol yield (0.48 ± 0.02 g g(-1)), theoretical yield (93 ± 3%), and extracellular invertase productivity (1,430 ± 50 IU l(-1) h(-1)), respectively, when fermenting 180 g sugars l(-1) in molasses medium at 43 °C in 300 m(3) working volume fermenter. Ethanol production was highly dependent on invertase production. Enthalpy (ΔH*) (32.27 kJ M(-1)) and entropy (ΔS*) (-202.88 J M(-1) K(-1)) values at 43 °C by the mutant MLD10 were significantly lower than those of ß-glucosidase production by a thermophilic mutant derivative of Thermomyces lanuginosus. These results confirmed the enhanced production of ethanol and invertase by this mutant derivative. These studies proved that mutant was significantly improved for ethanol production and was thermostable in nature. Lower fermentation time for ethanol production and maintenance of ethanol production rates (3.1 g l(-1) h(-1)) at higher temperature (43 °C) by this mutant could decrease the overall cost of fermentation process and increase the quality of ethanol production.

Evaluation of Antimicrobial Activity of Glucose Oxidase from Aspergillus niger EBL-A and Penicillium notatum

This work aimed to study the production and purification of glucose oxidase by Aspergillus niger and Penicillium notatum using corn steep liquor as the substrate and evaluate its antimicrobial activity for use in pharmaceutical and food industries. The enzyme was purified by ammonium sulfate precipitation (60-85%), DEAE-cellulose ion exchange and Sephadex G-200 size exclusion chromatography. The crude enzyme extracts of A. niger and P. notatum showed 2.32 and 5.53 U mg-1 specific activities, respectively, which after desalting was 15.52 and 12.05 U mg-1, and after ion exchange and gel filtration chromatography was 29.09 - 62 and 25.72 - 59.37 U mg-1 for A. niger and P. notatum, respectively. The antimicrobial activity was determined by disc diffusion method against selected microbial strains where glucose oxidase from A. niger showed anti-bacterial activity, while no fungicidal effects were shown by both A. niger and P. notatum glucose oxidases.

Kinetic characterization of glucose aerodehydrogenase from Aspergillus niger EMS-150-F after optimizing the dose of mutagen for enhanced production of enzyme

In the present study enhanced production of glucose aerodehydrogenase from Aspergillus niger has been achieved after optimizing the dose of chemical mutagen ethyl methane sulfonate (EMS) that has not been reported earlier. Different doses of mutagen were applied and a strain was developed basing upon the best production. The selected strain Aspergillus niger EMS-150-F was optimized for nutrient requirements in order to produce enzyme through fermentation and the results showed the best yield at 2% corn steep liquor (CSL), 36 hours fermentation time, pH 5, 30°C temperature, 0.3% KH2PO4, 0.3% urea and 0.06% CaCO3. The enzyme was then purified and resulted in 57.88 fold purification with 52.12% recovery. On kinetic characterization, the enzyme showed optimum activity at pH 6 and temperature 30°C. The Michaelis-Menton constants (Km, Vmax, Kcat and Kcat/Km) were 20 mM, 45.87 U mL-1, 1118.81 s-1 and 55.94 s-1 mM-1, respectively. The enzyme was found to be thermaly stable and the enthalpy and free energy showed an increase with increase in temperature and ΔS* was highly negative proving the enzyme from A. niger EMS-150-F resistant to temperature and showing a very little disorderliness.

Kinetic characterization of glucose aerodehydrogenase from Aspergillus niger EMS-150-F after optimizing the dose of mutagen for enhanced production of enzyme.

In the present study enhanced production of glucose aerodehydrogenase from Aspergillus niger has been achieved after optimizing the dose of chemical mutagen ethyl methane sulfonate (EMS) that has not been reported earlier. Different doses of mutagen were applied and a strain was developed basing upon the best production. The selected strain Aspergillus niger EMS-150-F was optimized for nutrient requirements in order to produce enzyme through fermentation and the results showed the best yield at 2% corn steep liquor (CSL), 36 hours fermentation time, pH 5, 30 °C temperature, 0.3% KH2PO4, 0.3% urea and 0.06% CaCO3. The enzyme was then purified and resulted in 57.88 fold purification with 52.12% recovery. On kinetic characterization, the enzyme showed optimum activity at pH 6 and temperature 30 °C. The Michaelis-Menton constants (K(m), Vmax, Kcat and Kcat/K(m)) were 20 mM, 45.87 U mL(-1), 1118.81 s(-1) and 55.94 s(-1) mM(-1), respectively. The enzyme was found to be thermally stable and the enthalpy and free energy showed an increase with increase in temperature and ΔS* was highly negative proving the enzyme from A. niger EMS-150-F resistant to temperature and showing a very little disorderliness.