The current study was undertaken to immobilize Kluveromyces lactis β-galactosidase on alginate coated magnesium oxide nanoparticles (ACMONPs). Transmission electron microscopy showed that MgO-NPs synthesized by wet chemical approach were of 27 nm size and spherical in shape. Excellent biocompatibility of sodium alginate resulted in 90% immobilization yield for β-galactosidase due to covalent attachment. Soluble and immobilized β-galactosidase exhibited its pH and temperature-optima at pH 7.0 and at 40 °C, respectively. However, the enzyme attached to ACMONPs enhanced its activity at higher and lower pH and temperature ranges, in contrast to enzyme in solution. ACMONPs bound enzyme displayed greater enzyme activity under the effect of galactose mediated product inhibition as well as in reusability experiment. Our study indicated that 88% lactose was hydrolyzed by β-galactosidase immobilized on ACMONPs at 40 °C as compared to 71% by soluble enzyme under identical temperature in controlled batch reactors. Hence, this nanomatrix can find its potential application in continuous packed-bed reactors for obtaining lactose-free dairy products.

Abstract Objective: To evaluate the effect of different preparations of fluoride gels on the salivary pH of albino rats. Material and Methods: This experimental study consisted of 40 Albino rats randomly divided into four equal groups. Group A was the control group and received no intervention. Experimental group B received a topical application of 0.2% sodium fluoride gel. Experimental group C received topical application of stannous fluoride gel 0.4%. Experimental group D received topical application of APF gel (1.23% acidulated phosphate fluoride gel). The different preparations of the gels were applied once daily for 4 minutes on the occlusal surface of the right maxillary molars for 14 days. Salivary pH values were recorded immediately after the application of gels with the help of pH paper on day 1 and day 14. Results: There was a significant difference in the pH level of groups B, C and D after 14 days of fluoride application (p < 0.05). The non-parametric Kruskal Wallis test was applied for the comparison between the groups. Conclusion: This study concluded that all the fluoride gels after administration caused the acidic pH of saliva with the most acidic effect produced by APF gel.

Resumo Sabe-se que os inibidores de serina protease (serpinas), uma superfamília de inibidores de protease, estão envolvidos em vários processos fisiológicos, como desenvolvimento, metamorfose e imunidade inata. Neste estudo, um cDNA de serpina de comprimento total, denominado Haserpin1, foi isolado da lagarta Helicoverpa armigera na cultura de algodão. A sequência de ADNc de Haserpin1 tem 1.176 nt de comprimento, com uma grelha de leitura aberta que codifica 391 aminoácidos; existe um éxon, mas nenhum íntron. O peso molecular previsto de Haserpin1 é de 43,53 kDa, com um ponto isoelétrico de 4,98. O InterProScan foi empregado para a caracterização funcional do Haserpin1, que revelou que o Haserpin1 contém motivos de assinatura altamente conservados, incluindo um loop central reativo (RCL) com uma região de dobradiça (E341-N350), a assinatura da serpina (F367-F375) e um local de clivagem previsto de P1-P1' (L357-S358), que são úteis para identificar serpinas. As transcrições de Haserpin1 foram expressas constitutivamente no corpo gordo, sugerindo que é o principal local para a síntese de serpinas. Durante os estágios de desenvolvimento, observou-se uma flutuação no nível de expressão de Haserpin1, com baixa expressão detectada no estágio larval do 5º ínstar. Por outro lado, detectou-se uma expressão relativamente alta no estágio pré-pupal, sugerindo que o Haserpin1 pode desempenhar um papel crítico no estágio errante de H. armigera. Embora a função detalhada dessa serpina (Haserpin1) precise ser elucidada, este estudo fornece uma perspectiva para a investigação funcional dos genes inibidores da serina protease.

Abstract Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341–N350), the serpin signature, (F367–F375) and a predicted P1–P1′ cleavage site (L357–S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.

ABSTRACT Salinity is a serious problem that limits crop growth and yield. The present study used plotting to evaluate 25 tomato (Solanum lycopersicum L.) accessions for various morpho-physiological parameters at the seedling stage and identify significantly salt-tolerant tomato lines at three different salinity levels. The pot experiment had a completely randomized design with three replicates in a factorial arrangement under greenhouse conditions. Tomato plants were exposed to 0 (control), 8, and 12 dS m-1 as NaCl stress at the seedling stage. The morpho-physiological traits, such as root and shoot length, root/shoot ratio, number of leaves, fresh and dry shoot weight, fresh and dry root weight, leaf area, Na+ and K+ concentrations, K+/Na+ ratio, and tolerance index, were recorded to examine salt tolerance. According to principal component analysis (PCA), there were six principal components (PCs) with Eigen values > 1 and 77.2% of total cumulative variability. The PC1 (24.3%) revealed the highest variability followed by PC2 (16.2%). Meanwhile, the PCA biplot and cluster heat map analyses indicated that Subarctic, Raad-Red, Naqeeb, Pakit, Tommy-Toe, and BL-1076 were salt-tolerant, whereas PBLA-1401, PB-017902, CLN-2413, BL-1078, BL-1174, and BL-1079 were the most susceptible accessions based on their performance under stress.

The present study investigates the synthesis of agarose nanoparticles (ANPs) and its surface modification by galactose for the immobilization of β-galactosidase. Galactose modified ANPs retained 91% enzyme activity upon immobilization. Optimum pH (4.5) and temperature (50 ºC) remains unchanged after immobilization. However, immobilized enzyme retained greater catalytic activity against lower and higher, pH and temperature ranges. Immobilized β-galactosidase retained 89% biocatalytic activity even at 4% galactose concentration as compared to enzyme in solution, and exhibited 81% activity even after seventh repeated uses. Immobilized enzyme hydrolyzed greater amount of lactose at higher temperatures as compared to β-galactosidase in solution, thereby suggesting its potential application in obtaining lactose-free dairy products at large scale.

ABSTRACT The presence of geneticdiversity is of great importance in improving wheat traits and developing strategies for optimal conservation of germplasm. Genetic diversity was assessed among common wheat cultivars using RAPD (Random Amplified Polymorphic DNA) markers at the Center of Agriculture, Biochemistry and Biotechnology (CABB), University of Agriculture, Faisalabad. RAPD primers were used among 14 Pakistani wheat cultivars, to screen the progenies and for the identification of the genes of interest. The polymorphic information content (PIC), was measured as the percentage of polymorphic fragments for all primers. A total of 583 bands(84% polymorphic) in all 14 wheat cultivars was amplified and discriminated all the wheat genotypes. The number of fragments amplified per primer ranged from 35 to 69 with an average of 48.52 fragments per primer averagely was observed. Population structure analysis anddandrogram showed distinct clustering among different wheat genotypes. Millat-11, Punjab-11, PBW-222 generated the maximum level of polymorphism, standing alone in the cluster while others are scatteredin different group.As a result, genetically numerous progenies are known, increasing the quality of sorts collections by broadening the genetic base of wheat cultivars. This study additionally indicates that RAPD markers allow quicker response and provide high throughputprocedure of accessions from a variety assortment to assess genetic diversity among wheat genotypes.

Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE), in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase) using a novel substrate, Banana Peels (BP) for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes.

BACKGROUND: In recent years, the desire to adopt a healthy diet has drawn attention to legume seeds and food products derived from them. Mash bean is an important legume crop used in Pakistan however a systematic mapping of the chemical composition of mash bean seeds is lacking. Therefore seeds of four mash bean (Vigna mungo (L.) Hepper, family Leguminoseae) cultivars (NARC-Mash-1, NARC-Mash-2, NARC-Mash-3, NARC-Mash-97) commonly consumed in Pakistan have been analyzed for their chemical composition, antioxidant potential and biological activities like inhibition of formation of advanced glycation end products (AGE) activity and tyrosinase inhibition activity. RESULTS: The investigated cultivars varied in terms of biochemical composition to various extents. Mineral composition indicated potassium and zinc in highest and lowest amounts respectively, in all cultivars. The amino acid profile in protein of these cultivars suggested cysteine is present in lowest quantity in all cultivars while fatty acid distribution pattern indicated unsaturated fatty acids as major fatty acids in all cultivars. All cultivars were found to be rich source of tocopherols and sterols. Fourier transform infrared spectroscopy (FTIR) fingerprints of seed flour and extracts indicated major functional groups such as polysaccharides, lipids, amides, amines and amino acids. Results indicated that all investigated cultivars possessed appreciable antioxidant potential. CONCLUSIONS: All cultivars are rich source of protein and possess sufficient content of dietary fiber, a balanced amino acid profile, low saturated fatty acids and antioxidant capacity that rationalizes many traditional uses of seeds of this crop besides its nutritional importance. The collected data will be useful for academic and corporate researchers, nutritionists and clinical dieticians as well as consumers. If proper attention is paid, it may become an important export commodity and may fetch considerable foreign exchange for Pakistan.

Adopting new technology always involves advantages and risks; Bt cotton (Gossypium hirsutum L.) is a new technology well known in developed countries for its many advantages, such as reduced pesticide application, better insect pest control, and higher lint yield. However, its success in developing countries is still a question mark. Global adoption of Bt cotton has risen dramatically from 0.76 million ha when introduced in 1996 to 7.85 million ha in the 2005 cotton-growing season where 54% of the cotton crops in the USA, 76% in China, and 80% in Australia were grown with single or multiple Bt genes. Bollworms are serious cotton pests causing 30-40% yield reduction in Pakistan and 20-66% potential crop losses in India. The major advances shown in this review include: (1) Evolution of Bt cotton may prove to be a green revolution to enhance cotton yield; (2) adoption of Bt cotton by farmers is increasing due to its beneficial environmental effects by reducing pesticide application: however, a high seed price has compelled farmers to use illegal non-approved Bt causing huge damage to crops because of low tolerance to insect pests; and (3) some factors responsible for changes in the efficiency of the Bt gene and Bt cotton yield include internal phenology (genetics), atmospheric changes (CO2 concentration), nutrition, insect pests, boll distribution pattern, disease and nematodes, removal of fruiting branch and/or floral bud, introduction of Bt gene, and terpenoids and tannin production in the plant body.

La adopción de nueva tecnología siempre involucra ventajas y riesgos; algodón Bt (Gossypium hirsutum L.) es una nueva tecnología bien conocida en países desarrollados por muchas ventajas como reducida aplicación de pesticidas, mejor control de insectos plaga, y mayor producción de fibra, pero su éxito en países en desarrollo aún conlleva dudas. La adopción global de algodón Bt ha aumentado dramáticamente de 0,76 millones de hectáreas en su introducción en 1996 a 7,85 millones de hectáreas en la estación de cultivo de algodón 2005, 54% de cultivos de algodón en EE.UU., 76% en China, y 80% en Australia se cultivaron con genes Bt únicos o múltiples. Los gusanos del algodonero son plagas graves del algodón que causan 30-40% de reducción en rendimiento en Paquistán, y 20-60% pérdidas potenciales en India. Las principales ventajas mostradas en esta revisión incluyen: (1) Evolución de algodón Bt puede probar revolución verde en el aumento de rendimiento de algodón; (2) adopción de algodón Bt por agricultores está aumentando debido a sus efectos beneficiosos en el medioambiente al reducir aplicación de pesticidas, pero el alto precio de la semilla ha obligado a los agricultores a usar semilla Bt ilegal no aprobada que causó gran dano al cultivo debido a baja tolerancia a insectos plaga; y (3) algunos factores responsables de cambios en eficiencia de genes Bt y por lo tanto de rendimiento del algodón Bt incluyen fenología interna (genética), cambios atmosféricos (concentración de CO2), nutrición, insectos plaga, patrón de distribución de gusanos, enfermedades y nematodos, remoción de ramas fructificantes y/o yemas florales, introducción de genes Bt y producción de terpenoides, taninos, etc., dentro del cuerpo de la planta.