Indigenous Ethiopian okra (Abelmoschus esculentus) mucilage: A novel ingredient with functional and antioxidant properties.

Functional and antioxidant properties of mucilage extracted from the pods of eight okra accessions grown in Benishangul-Gumuz region, Western Ethiopia, were evaluated. This study had shown that the mucilage contents of the pods of eight okra accessions ranged from 1.25 to 3.45 g/100 g. Functional properties of the mucilage of okra pods varied significantly (p < .05) and had respective ranges of bulk density of 0.58-0.64 g/ml; water absorption capacity of 2.45-4.60 ml/g; oil absorption capacity of 0.02-3.64 ml/g; emulsifying capacity of 42.22%-74.45%; emulsion stability of 42.22%-74.45%; foaming capacity of 50.51%-62.50%, and foam stability of 36.04%-54.35%. Total phenolic and flavonoid contents of the mucilage of the pods of okra accessions ranged from 4.66 to 49.93 mg GAE/g and 8.18-18.72 mg CE/g, respectively. The effective concentration (EC 50) values (mg/ml) of mucilage of okra pods varied from 3.15 to 6.60 and 1.10 to 1.85 for DPPH scavenging and metal-chelating activity, respectively. The study revealed that the mucilage of the pods of okra accessions was found to exhibit good functional properties and can offer a great potential in various food systems. Particularly, mucilage of the pods from OPA#5 and OPA#7 had desirable water and oil absorption capacities, whereas the mucilage of accession OPA#1 and OPA#6 had high emulsifying and foaming properties. The results also demonstrated that okra pod mucilage had potential sources of natural antioxidant.

Microbiological, biochemical, and functional aspects of sugary kefir fermentation - A review.

Sugary kefir beverage is produce by fermenting raw sugar solution with kefir grains, the latter consisting of polysaccharide and microorganisms. This beverage, with great consumption in countries such as USA, Japan, France, and Brazil, represents a promising market to functional cultured drinks. This paper reviews the microbial diversity and interaction, kinetics, safety, and bioactivities of sugary kefir fermentation. The literature reviewed here demonstrates that sugary kefir possesses a similar microbial association relative to traditional milk kefir fermentation, especially among lactic acid bacteria and yeast species, such as Lactobacillus, Leuconostoc, Kluyveromyces, Pichia, and Saccharomyces. However, a selective pressure at species level is generally observed, as, for example, the stimulation of Saccharomyces species metabolism, leading to a high content of alcohol in the final product. This also seems to stimulate the growth of acetic acid bacteria that benefit of increased ethanol production to acetic acid metabolism. Existing reports have suggested important bioactivities associated with sugary kefir beverage consumption, such as antimicrobial, antiedematogenic, anti-inflammatory, antioxidant, cicatrizing, and healing activities. Other alternative non-dairy substrates, such as fruits, vegetables, and molasses, have also been tested for adaptation of kefir grains and production of functional beverages with distinct sensory characteristics. This diversification is of crucial importance for the production of new probiotic products to provide people with special needs (lactose intolerance) and vegan consumers.

Proximate, mineral, and antinutrient compositions of indigenous Okra (Abelmoschus esculentus) pod accessions: implications for mineral bioavailability.

The promotion and consumption of indigenous vegetables could help to mitigate food insecurity and alleviate malnutrition in developing countries. Nutrient and antinutrient compositions of eight accessions of Okra Pods were investigated. Molar ratios and mineral bioavailability of Okra pod accessions were also calculated and compared to the critical values to predict the implications for mineral bioavailability. Proximate and mineral composition of Okra pod accessions were determined using standard methods of Association of Official Analytical Chemists. The result of the study revealed that the proximate composition (g/100 g) in dry weight basis was significantly (P < 0.05) varied and ranged: moisture/dry matter 9.69-13.33, crude protein 10.25-26.16, crude fat 0.56-2.49, crude fiber 11.97-29.93, crude ash 5.37-11.30, utilizable carbohydrate 36.66-50.97, and gross energy 197.26-245.55 kcal/100 g. The mineral concentrations (mg/100 g) were also significantly (P < 0.05) varied and ranged: calcium (111.11-311.95), Iron (18.30-36.68), potassium (122.59-318.20), zinc (3.83-6.31), phosphorus (25.62-59.72), and sodium (3.33-8.31) on dry weight bases. The Okra Pods of "OPA#6" accession contained significantly higher amounts of crude protein, total ash, crude fat, calcium, iron, and zinc than all other accessions evaluated in this study. The results of antinutrients analysis showed that, except phytate, tannin, and oxalate contents of all the accessions were significantly (P < 0.05) varied. The range of phytate, tannin, and oxalate contents (mg/100 g) for Okra pod accessions studied were as follows: 0.83-0.87, 4.93-9.90, and 0.04-0.53, respectively. The calculated molar ratios of phytate: calcium, phytate: iron, phytate: zinc, oxalate: calcium and [Phytate][Calcium]/[Zinc] were below the critical value and this indicate that the bioavailability of calcium, iron, and zinc in these accessions could be high. The results of the study revealed that Okra pod contain appreciable amount of vital nutrients like protein, fiber, calcium, iron, and zinc and low in antinutrient contents with high mineral bioavailability. Therefore, increase in the production and consumption of these nutrient-rich indigenous Okra pods will help to supplement/formulate the diets and alleviate the problems associated with malnutrition in the country.

Effect of different pretreatments on delignification pattern and enzymatic hydrolysability of miscanthus, oil palm biomass and typha grass.

Cumulative pretreatments methods were evaluated for delignification ability and enzymatic digestibility using miscanthus (M×G), empty palm fruit bunch (EFB) and typha grass as feedstocks. Despite their close chemical composition, the three feedstocks unveiled quite different behavior under the same condition of pretreatment. Characterization of ethanol organosol lignins extracted from the three feedstocks by (13)C NMR and FTIR revealed information concerning S/G/H ratios which was important to rationalize the differences among the feedstock behavior. The S/G/H ratios for MxG, EFB and typha, were established to levels of ~52/44/4, ~68/30/2 and ~46/27/27 respectively. The xylans hydrolytic susceptibility were a major cause of difference in behavior of feedstock during the pretreatment process. The influence of the presence of naphthol during autohydrolysis on the delignification ability was studied. A good relationship was observed between S+H/G ratio and the scavenging effect of naphthol.

Comparison of some new pretreatment methods for second generation bioethanol production from wheat straw and water hyacinth.

Pretreatment of lignocellulosic residues like water hyacinth (WH) and wheat straw (WS) using crude glycerol (CG) and ionic liquids (IL) pretreatment was evaluated and compared with conventional dilute acid pretreatment (DAT) in terms of enzymatic hydrolysis yield and fermentation yield of pretreated samples. In the case of WS, 1-butyl-3-methylimidazolium acetate pretreatment was found to be the best method. The hydrolysis yields of glucose and total reducing sugars were 2.1 and 3.3 times respectively higher by IL pretreatment than DAT, while it was 1.4 and 1.9 times respectively higher with CG pretreatment. For WH sample, CG pretreatment was as effective as DAT and more effective than IL pretreatment regarding hydrolysis yield. The fermentation inhibition was not noticeable with both types of pretreatment methods and feedstocks. Besides, CG pretreatment was found as effective as pure glycerol pretreatment for both feedstocks. This opens up an attractive economic route for the utilization of CG.

The Stability of rRNA in Heat-killed Salmonella enterica Cells and Its Detection by Fluorescent In Situ Hybridisation (FISH).

Differentiation of viable cells from non-viable cells is a major concern in the detection of foodborne microbial pathogens. Fluorescent in situ hybridisation (FISH) has been utilised as a promising method in this regard. The ability of FISH to differentiate viable cells from non-viable cells depends on the rapid degradation of rRNA in non-viable cells. In our work, Salmonella enterica that were heat-killed at 80°C, 100°C and 121°C were examined for the presence of rRNA using FISH at various times ranging from 5 minutes to 48 hours after heat treatment. rRNA was detected by FISH in heat-killed bacteria for 12 hours, 3 hours and 1 hour after treatment at 80°C, 100°C and 121°C, respectively. These results indicate that there is a correlation between temperature and stability of rRNA in heat-killed bacteria. In conclusion, FISH was determined to be a suitable method for differentiation of viable cells from non-viable cells, especially for samples subjected to extreme heat.

Effect of alpha-ketoglutarate on monoclonal antibody production of hybridoma cell lines in serum-free and serum-containing medium.

Process development and optimization for increase population growth and protein productivity in mammalian cell culture have been studied for many years. In this study, the behavior of hybridoma cells was investigated using six-well micro-titer plate systems with a working volume of 4 ml. Mouse hybridoma cell lines D2 and 2C83G2 were seeded in serum-free and serum-containing media and cultured for 8 days. alpha-Ketoglutarate is an integral component of the tricarboxylic acid (TCA) cycle and is produced from glutamine via glutamate. To study its effect on cell growth, metabolism, and monoclonal antibody (mAb) production, 2 mM alpha-ketoglutarate (pH 7.2) was added in both media at the beginning of the cultivation and in another set after 72 h. High cell density was observed in D2 cell culturing in serum-free medium, while 2C83G2 cell line showed high cell density in serum-containing medium. However, both cell lines cultured in serum-free medium gave viability above 70% when grown for 8 days. The supplement of 2 mM alpha-ketoglutarate supported cell growth and mAb production of both hybridoma cell lines in serum-free and serum-containing medium. The addition of alpha-ketoglutarate at the beginning of the batch cultivation gave better result in cell growth and mAb production as compared to alpha-ketoglutarate supplementation after 72 h. However, addition after 72 h was better than no addition at all. This indicates that alpha-ketoglutarate have a positive effect on production and release of antibody.

Ultrasound pretreatment of cassava chip slurry to enhance sugar release for subsequent ethanol production.

The use of ultrasound pretreatment to enhance liquefaction and saccharification of cassava chips was investigated. Cassava chip slurry samples were subjected to sonication for 10-40 s at three power levels of low (2 W/mL), medium (5 W/mL), and high (8 W/mL). The samples were simultaneously exposed to enzymes to convert starch into glucose. The cassava particle size declined nearly 40-fold following ultrasonic pretreatment at high power input. Scanning electron micrographs of both unsonicated (control) and sonicated samples showed disruption of fibrous material in cassava chips but did not affect the granular structure of starch. Reducing sugar release improved in direct proportion to the power input and sonication time. The reducing sugar increase was as much as 180% with respect to the control groups. The slurry samples with enzyme addition during sonication resulted in better reducing sugar release than the samples with enzyme addition after sonication. The heat generated during sonication below starch gelatinization temperature apparently had no effect on the reducing sugar release. The reducing sugar yield and energy efficiency of ultrasound pretreated samples increased with total solids (TS) contents. The highest reducing sugar yield of 22 g/100 g of sample and efficiency of 323% were obtained for cassava slurry with 25% TS at high power. The reducing sugar yield at the completion of reaction (R(infinity)) were over twofold higher compared to the control groups. The integration of ultrasound into a cassava-based ethanol plant may significantly improve the overall ethanol yield.

Three-dimensional culture for monoclonal antibody production by hybridoma cells immobilized in macroporous gel particles.

Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on multiwell plate surfaces was also investigated. The effect of three different medium such as basal medium Dulbecco's modified Eagle's medium (D-MEM) containing L-glutamine or L-glutamine + 2 mM alpha-ketoglutarate or L-alanyl-glutamine (GlutaMAXtrade mark) was studied prior to its use in 3D cultivation. The kinetics of cell growth in basal medium containing L-glutamine + alpha-ketoglutarate was similar to cells grown on GlutaMAX containing medium, whereas D-MEM containing L-glutamine showed lower productivity. With the maximal viable cell density (6.85 x 10(6) cells mL(-1)) and highest specific mAb production rate (3.9 mug mL(-1) 10(-4) viable cell day(-1)), D-MEM-GlutaMAX was further selected for 3D cultivation. Cells in MGPs were able to grow and secrete antibody for 30 days in packed-bed batch reactor, before a fresh medium reservoir was replaced. After being supplied with fresh medium, cells again showed continuous growth for another 30 days with mAb production efficiency of 50%. These results demonstrate that MGPs can be used efficiently as supporting carrier for long-term monoclonal antibody production.

Monoclonal antibody production using a new supermacroporous cryogel bioreactor.

A supermacroporous cryogel bioreactor has been developed to culture hybridoma cells for long-term continuous production of monoclonal antibodies (mAb). Hybridoma clone M2139, secreting antibodies against J1 epitope (GERGAAGIAGPK; amino acids, 551-564) of collagen type II, are immobilized in the porous bed matrix of a cryogel column (10 mL bed volume). The cells got attached to the matrix within 48 h after inoculation and grew as a confluent sheet inside the cryogel matrix. Cells were in the lag phase for 15 days and secreted mAb into the circulation medium. Glucose consumption and lactic acid production were also monitored, and during the exponential phase (approximately 20 days), the hybridoma cell line consumed 0.75 mM day-1 glucose, produced 2.48 mM day-1 lactic acid, and produced 6.5 microg mL-1 day-1 mAb during the exponential phase. The mAb concentration reached 130 microg mL-1 after continuous run of the cryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, which was three times greater than the mAb yield obtained from T-flask batch cultivation. Even after the exchange of medium reservoir, cells in the cryogel column were still active and had relatively stable mAb production for an extended period of time. The bioreactor was operated continuously for 55 days without any contamination. The results from ELISA as well as arthritis experiments demonstrate that the antibodies secreted by cells grown on the cryogel column did not differ from antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus, supermacroporous cryogels can be useful as a supporting material for productive hybridoma cell culture. Cells were found to be viable inside the porous matrix of the cryogel during the study period and secreted antibodies continuously. The antibodies thus obtained from the cryogel reactor were found to be functionally active in vivo, as demonstrated by their capacity to induce arthritis in mice.