A bacterial endo-polygalacturonase (endo-PGase) gene from the plant pathogen Pectobacterium carotovorum was cloned into pGAPZαA vector and constitutively expressed in Pichia pastoris. The recombinant endoPGase secreted by the Pichia clone showed a 1.7 fold increase when the culture medium included glycerol in replacement of glucose as the carbon source. The enzyme had optimum activity at pH 5.5 and 40 °C with stability between pH 5.0 and 8.0 and at temperatures up to 50 °C. The enzyme activity was enhanced by 41% with the addition of 1 mM Co ++ , and inhibited by Fe ++ with a 63% reduction. The mode of the enzyme action showed internal cleavage of α-1,4 glycoside bonds of polygalacturonic acid and citrus peel pectin. Trigalacturonate and hexagalacturonate were the main hydrolysis products, with a yield of 0.44±0.01 and 0.21±0.01 mg released per mg polygalacturonic acid substrate, respectively. This represents the first report of a microbial endo-PGase that produced trimer and hexamer uniquely as the end products of hydrolysis, in contrast to mixtures of mono-, di-, and trigalacturonates commonly observed for the action of fungal enzymes. Pectic oligosaccharides generated from native carbohydrate polymers offer the potential application as building blocks for value-added products.

Pectinolytic enzymes or pectinases are synthesized naturally by numerous microbes and plants. These enzymes degrade various kinds of pectin which exist as the major component of the cell wall in plants. A pectinase gene encoding endo-polygalacturonase (endo-PGase) enzyme was isolated from Pectobacterium carotovorum a plant pathogenic strain of bacteria and successfully cloned into a secretion vector pHT43 having σA-dependent promoter for heterologous expression in Bacillus subtilis (WB800N).The desired PCR product was 1209bp which encoded an open reading frame of 402 amino acids. Recombinant proteins showed an estimated molecular weight of 48 kDa confirmed by sodium dodecyl sulphate–polyacrylamide-gel electrophoresis. Transformed B. subtilis competent cells harbouring the engineered pHT43 vector with the foreign endo-PGase gene were cultured in 2X-yeast extract tryptone medium and subsequently screened for enzyme activity at various temperatures and pH ranges. Optimal activity of recombinant endo-PGase was found at 40°C and pH 5.0. To assay the catalytic effect of metal ions, the recombinant enzyme was incubated with 1 mM concentration of various metal ions. Potassium chloride increased the enzyme activity while EDTA, Zn++ and Ca++, strongly inhibited the activity. The chromatographic analysis of enzymatic hydrolysates of polygalacturonic acid (PGA) and pectin substrates using HPLC and TLC revealed tri and tetra-galacturonates as the end products of recombinant endo-PGase hydrolysis. Conclusively, endo-PGase gene from the plant pathogenic strain was successfully expressed in Bacillus subtilis for the first time using pHT43 expression vector and could be assessed for enzyme production using a very simple medium with IPTG induction. These findings proposed that the Bacillus expression system might be safer to escape endotoxins for commercial enzyme production as compared to yeast and fungi. Additionally, the hydrolysis products generated by the recombinant endo-PGase activity offer their useful applications in food and beverage industry for quality products.

Purpose Recent advances in nanotechnology have given rise to the potential utilization of nanoparticles as food, nano-medicine/biomedicines. Patient The study aimed to investigate the effects of nano-zinc oxide (nano-zinc) on the bio-assimilation of mineral (Zn) in mice, aged 3–6 weeks. Methods ZnO nanoparticles were added to the basal diet as a supplement at amounts of 0.07, 0.14 and 0.21 mg/kg. The synthesized material was characterized by Fourier transform infrared spectrophotometer, particle size, scanning electron microscope, Thermogravimetric Analysis Thermal, X-ray diffraction spectrophotometer and Zeta potential. Results In-vitro bioavailability of synthesized group ZnO (120 nm) was 43%, whereas for standard group ZnO (50 nm) was reported as 55%. In-vivo bioavailability of zinc oxide illustrated the maximum absorption level compared with the control. In-vivo toxicity was characterized as damage done to the liver and spleen tissues with a high dose of 0.21 mg/kg, while smaller doses indicated no toxic effects. Conclusion The study provided important insights on the toxicological effects of ZnO nanoparticles, depending on dose rate and bio-assimilation, as well as particles, under various conditions (in-vitro and in-vivo). These findings will motivate further detailed research on nano-based medicine for alleviating malnutrition conditions.

Globally, over-reliance on major food crops (wheat, rice and maize) has led to food basket’s shrinking, while climate change, environmental pollution and deteriorating soil fertility demand the cultivation of less exhaustive but nutritious grasses. Unlike neglected grasses (grass species restricted to their centres of origin and only grown at the subsistence level), many underutilized grasses (grass species whose yield or usability potential remains unrealized) are resistant and resilient to abiotic stresses and have multiple uses including food (Coix lacryma-jobi), feed (Eragrostis amabilis and Cynodon dactylon), esthetic value (Miscanthus sinensis and Imperata cylindrica), renewable energy production (Spartina pectinata and Andropogon gerardii Vitman) and contribution to ecosystem services (Saccharum spontaneum). Lack of agricultural market globalization, urbanization and prevalence of large commercial enterprises that favor major grasses trade, improved communication means that promoted specialization in favor of established crops, scant planting material of underutilized grasses and fewer research on their production technology and products development are the prime challenges posed to underutilized grasses promotion. Integration of agronomic research with novel plant protection measures and plant breeding and molecular genetics approaches for developing biotic and abiotic stresses tolerant cultivars along with the development of commercially attractive food products hold the future key for promoting underutilized grasses for supplanting food security and sustainably multiplying economic outcomes.

Red kidney bean protein isolate was prepared and incorporated into wheat flour at levels of 0, 5, 10, 15, and 20% to prepare protein-enriched cookies which were then evaluated for various quality attributes. Nutritional evaluation of cookies showed that the supplementation of bean protein isolate increased the protein content of the cookies from 7.87% in T₁ (control or wheat flour cookies) to 16.92% in T₆(cookies supplemented with 20% bean protein isolate), which indicated a 115% increase in protein content. The physical characteristics of cookies such as width, thickness and spread factors differed significantly. Sensory attributes of all the supplemented cookies were within the acceptable range but cookies supplemented with 20% level of protein isolate were less preferred due to slightly darker colour and harder texture. Conclusively, the red kidney bean protein isolate can be successfully utilised for the production of high-protein cookies. Such high-protein products can be advantageously utilised to cope with the problem of protein-energy malnutrition. Moreover, such nutritious products can also be used as emergency foods during natural disasters as well as during war times.

Lactobacillus is an important class of Gram-positive, non-spore-forming bacteria for food industrial applications. The genus Lactobacillus is a potential candidate in fermentation technology for the production of fermented food, feed, and pharmaceutical products. The diverse features of Lactobacilli based on their capability to produce acids, enzymes, bacteriocins by fermenting carbohydrates. Lactobacilli have probiotic potential and therefore applied in dairy [cheese, yoghurt, fermented milk] and nondairy products such as sausages, juices as well as in animal feed in the form of starter culture. Among Lactobacilli, lactic acid-producing bacteria are applied as starter cultures in a variety of fermented foods. Lactobacilli are the natural microflora of the gastrointestinal tract and play a beneficial role against infections. The ability of Lactobacilli to produce bacteriocins and other antifungal compound lead to the development of bioprotective cultures for use in different foods. Bacteriocins has wide applications in food industries for preventing the attack of foodborne pathogens and for manufacturing active packaging materials. This chapter aimed to review significant industrial applications of Lactobacilli with specified strains and also starter cultures with their potential beneficial effects are engrossed. The chapter highlights the commercial applications of Lactobacilli in the food, feed, wine and pharmaceutical industries.

Endopolygalacturonase (EndoPGase) is one of the crucial pectinases belonging to the class of carbohydrase. The catalytic action of EndoPGase captivate the attention for production of this extremely valuable catalyst of industrial sector. The main focus was to ascertain a potential bacterial candidate for endoPGase production. The isolated bacterial strain was further identified by 16S rDNA gene sequencing. A genomic library was constructed by using Lambda ZAP II vector system to investigate the pectinolytic potential of the expressed genes. The parameters for enzyme biosynthesis were optimized by single as well as multiple factor approach at a time. The results of our investigation led to the identification of a potent strain of Bacillus subtilis NR2. The strain was found active for pectic enzyme activity under shaking- flask fermentation at pH 5.0 and 50 °C temperature of incubation. Among all monomeric and polymeric substrates, citrus pectin followed by wheat bran was considered the best enzyme inducers at 1 % concentration. Moreover, an increasing trend in enzyme activity was observed with the increasing inducer concentration. The combined effect of three variables (pH, substrates, and substrate concentration) was explored by response surface methodology involving Box Benken Design (BBD). The study concluded that the soil isolated B. subtilis can be utilized for commercial scale pectinase enzyme production.

Pectinolytic enzymes [pectinases] produced by microbes are highly important for their biotechnological use in processing of vegetables and fruits beverages and use in pulp and paper industry. A pectinases, namely endo-polygalacturonase [endo-PGase], encoding gene isolated from Pectobacterium carotovorum, a plant pathogenic strain of bacteria was successfully cloned into a secretion vector pHT43 having σ?-dependent promoter P grac . For enhanced expression analysis, competent cells of Bacillus subtilis (WB800N) were prepared at stationary phase using high salt medium. The recombinant B. subtilis competent cells, harboring the engineered pHT43 with the endo-PGase gene were cultured in 2X-yeast extract tryptone medium. The recombinant endo-PGase enzyme was secreted directly into the medium after 72 hours of the first IPTG induction. The recombinant endo-PGase was screened for its activity at various temperatures and pH ranges. Optimal activity was found at pH 5.0 and a temperature of 40°C with a stability ranging from pH 5.0-9.0. For detection of metal ion effect, recombinant enzyme was incubated with 1mM concentration of; Ca ++ , Mg ++ , Zn ++ , EDTA, K ++ for 45 minutes. Resultantly, Ca ++ , EDTA and Zn ++ strongly inhibited the enzyme activity. The chromatographic analysis of enzymatic hydrolysate of polygalacturonic acid [PGA] and pectin substrates using HPLC and TLC revealed that tri and tetra-galacturonates were the end products of hydrolysis. The study led to the conclusion that endo-PGase gene from the plant pathogenic strain was successfully expressed in Bacillus subtilis and assessed for enzyme production using a very simple medium with IPTG induction. These findings proposed that the Bacillus expression system might be safe for commercial enzyme production as compared to yeast and fungi to escape endotoxins.