Medium composition effects on growth kinetic of cordyceps militaris cells using agar plate method

Mushroom Cordyceps, as one of the most well known funguses with numerous bioactive compounds possess therapeutic actions; many years used as medicinal food particularly in China and Japan. Interestingly, the long history of mushroom as a therapeutic agent is not far from its role as a food component among Asian people specifically in Traditional Chinese Medicine (TCM). It has been cultivated naturally or in artificial media. Fungal mycelia contain adenosine, cordycepin, and polysaccharides, which are responsible for its biological activities. Cordycepin is the best-known and most potent mushroom-derived substances possessing anticancer, antitumor, antiviral, antihypertensive, immune response stimulating effects, blood lipid lowering effects and several other immunomodulating activities. Therefore, it can be a potential alternative or supplement to chemotherapy in order to treat or accelerate the treatment efficiency on the different types of human related cancer diseases. However, high quality and large-scale production of bioactive products from mushroom Cordyceps are the issues need to be investigated. Thus, optimization of cultural conditions such as medium composition and also the type of components used in medium are two essential factors, which are so effective on the acceleration of the product formation by the cells. In this study, optimized solid state cultivation of fungal mycelia cells using modified potato dextrose agar (PDA) medium culture supplemented with specific amount of malt extract (ME) together with yeast extract (YE) was investigated. The mycelial growth diameter was monitored during 21 days of cultivation time using two series of experiments including 2, 4, 6 and 8 g of ME and 6 g of ME with 0, 2, 4 and 6 g/L of YE in the PDA medium culture, respectively. Results illustrated the highest mycelial growth diameter around 7.5 cm using medium composed of PDA supplemented with 6 g/L ME and 4 g/L. Further investigations are now undertaken in our laboratories to clarify the effects of the new medium presented here on the bioactive metabolite formation (e.g. cordycepin)

Bioprocess development for anaerobic cultivation of probiotic bacteria bifidobacterium longum for high cell mass production

Bifidobacteria are used as probiotics mainly in the dairy industry as cell suspensions or as freeze-dried additives. Bifidobacterium longum (B. longum) are important in maintaining general health. The potential health benefits of B. longumto the human have led to their wide application in dairy products and food additives. Fastidious anaerobic growth of B. longum plus organic acids production as their byproductsgive some restriction in their growth as high cell mass become major concern. Therefore, the goal of this research is to select suitable medium as production media and optimization of the medium component for high cell mass production of B. longum by using one-factor-at-a-time (OFAT). Among nine growth media were evaluated to determine their suitability for high cell mass of B. longum, the best medium was yielded a cell mass of 4.5 g L-1 in shake study with the main components are glucose (20 g L-1), yeast extract (5 g L-1), meat extract (10 g L-1) and peptone (10 g L-1).Then, five different carbon sources which were glucose, lactose, sucrose, mannitol, and glycerol were screened in shake flasks and the best carbon source that contribute to the highest cell mass was glucose. Moreover, application of highly nutritious and costly nitrogen sources was incomplete application for industrial scale. Therefore, in this study peptone was found to be the best nitrogen source after screened with two others different nitrogen sources which were yeast extract and meat extract. Optimization by classical approaches achieving the maximum cell mass of 5.8 gL-1increased up to 28.88 % when compared with un-optimized media. Finally, batch cultivation was conducted in 16-L semi-scale bioreactor using the new formulated optimized medium under controlled and uncontrolled pH conditions at 37°C for 72 hours under anaerobic cultivation. It showed that under controlled pH, the maximal cell mass obtained in batch cultures was 13.8 gL-1 when compared with uncontrolled pH which only 6.8 gL-1with the percentage difference of 67.96 %. Thus the batch cultivation under controlled pH is the most suitable cultivation strategy for high cell mass production for industrialization of this bioprocess

Antibacterial activity of zingiber officinale and zingiber zerumbet by using Turbo Extractor Distillator (TED)

Zingiber officinale (ginger) and Zingiber zerumbet (lempoyang) are belong to Zingiberaceae family. These plants have been traditionally used as a treatment for stomach problems, nausea, vomiting, epilepsy, sore throat, muscular pains and several other disorders. Essential oils from both plants were investigated for their efficacy on antibacterial activity against two Gram positive (Staphylococcus aureus, ATCC 25923 and Bacillus cereus, ATCC 11778) and two Gram negative (Pseudomonas aeruginosa, ATCC 27853 and Escherichia coli, ATCC 35218) bacteria species using the disc diffusion assay. A zone of inhibition was compared with the standard antibiotic chloramphenicol (10 µg/disc), whilst a blank disc impregnated with the methanol was used as negative control. At concentration 20µl/disc, Zingiber officinale essential oils produced zone of inhibition ranging from 16mm to 36mm, while Zingiber zerumbet essential oils produced zone inhibition ranging from 11mm to 14mm. From these findings, Zingiber officinale essential oil inhibited the growth of all tested bacteria with large zone of inhibition. The most susceptible bacteria was Bacillus cereus while the lowest was Pseudomonas aeruginosa. It can be concluded that, Zingiber officinale and Zingiber zerumbet essential oils might provide potential therapeutic agents against bacterial infection

New formulation of production media for submerged cultivation of aspergillus niger for production of pectinase

Pectinase is a generic term that used from derivation of the pectin. Pectin is a complex class of carbohydrates polymer which composed of members galacturonic acid that linked through the a-1-4 glycosidic linkage and it is widely found in the primary cell walls or at the middle lamella of higher plants . Furthermore, Among different biofactories of pectinases, the filamentous fungi provide a potentially high yielding and relatively cheap option and the genus of Aspergillus has been used with a success as a production host . Therefore, the objective of this research is to develop industrial production media and a cultivation strategy for the production and secretion of pectinases in a semi-industrial scale by A. niger. In this study, the submerged cultivation was chose as a cultivation strategy for the production and secretion of pectinase in a semi-industrial scale by A. niger

Probiotication of Punica granatum (pomegranate) juice by lactobacillus plantarum

Fruit juice enriched with probiotics is increasingly accepted nowadays, mainly due to its health benefit for digestive system. In particular, probioticated fruit juice is the good choice for those who are having lactose intolerant problem from milk based drinks. In the present study, the whole fruit of Punica granatum (pomegranate) has been probioticated with Lactobacillus plantarum at different fermentation temperatures (22ºC, 30ºC and 35ºC). The growth rate of L. plantarum has been monitored based on the optical density and acidity of the broth culture at 24 hours of time interval for 72 hours. The bacterial growth in the pomegranate juice was predicted by measuring absorbance at 600 nm spectrophotometrically and pH value by a pH meter. There was an increasing trend in the bacterial growth of L. plantarumincubated at 35ºC compared to other temperatures at 22ºC and 30ºC. The results also indicated there was no significant changes on pH during the fermentation as the bacterial strain was in adaptation process with the new medium and conditions. Meanwhile, the antioxidant assay showed that probiotication of pomegranate juice by L. plantarum significantly increased the radical scavenging activity. The pomegranate juice was shown to be a suitable substrate for L. plantarum cultivation at 35ºC

Lactobacillus acidophilus and non-digestible carbohydrates: a review

In the recent years, lactic acid bacteria species such as Lactobacillus are considering one of the important species of probiotics used in the food processing sector to produce fermented products and play a significant role for the transformation and preservation of food products. Besides, there is a huge exploration of new molecules that promote health and exhibit potential for technological applications such as non-digestible carbohydrates. The non-digestible carbohydrates provide various health benefits such as balancing and sustaining the microbiota in the intestine and increasing the production of short chain fatty acids (SCFA). The aim of this review is to review some types of non-digestible carbohydrates as an enhancer for the growth of probiotics. These compounds can help in improving many characteristics of food such as sensory and textural properties

Optimization of polysaccharide productio by Lactobacillus kefiranofaciens using response surface methdology

Kefiran is an exopolysaccharides produced by Lactobacillus kefiranofaciens which was isolated from kefir grains. Kefiran has wide applications mainly in food and pharmaceutical industries. Growth and kefiran production of L. kefiranofaciens can be significantly enhanced by using mixed culture technique. Saccharomyces cerevisiae in this study was used to enhance the kefiran production by reducing lactic acid accumulation in the cultivation medium. The statistical analysis was used for optimization study by using response surface methodology based on Box-Behnken design. The interaction was studied between three different levels of variables that were lactose, yeast extract and phosphate. A second order polynomial model was used to correlate the factors. The model was found to be significant. The optimum concentration of lactose, yeast extract and phosphate obtained using statistical media optimization is 59.09 g L-1, 8.69 g L-1and 0.5 g L-1 respectively. Kefiran production in optimized medium was 0.97 g L-1 which gave an increase in kefiran production up to 42.65 % compared to the un-optimized medium which was only 0.68 g L-1 of kefiran

Production of extracellular thermostable recombinant phytase by Escherichia Coli B121 (DE3) when glycerol as carbon source and induced with lactose

Phosphorus content in plants in the form of phytic acid (myo-inositol hexakisphosphate) is also known as phytate. It serves as a primary depository form of high-energy hosphoryl groups and several divalent cations, Phytase is a type of phosphatase enzyme that catalyzes the degradation of phytic acid and undigested organic phosphorus to be released as useable form of inorganic phosphorus. In this study, a recombinant Escherichia colt BL21 (DE3) which harbouringthermostablephytase gene from Bacillus sp.MD2. It's known that IPTG as inducer is toxic to the cells and costly.Therefore, lactose has been used as an alternativeinducer in fermentation of recombinant E.coli. Unfortunately, lactose can be metabolized as carbon source and contribute to increase the metabolic overflow due to excess carbon sources during induction period which lead s to acetic acid accumulation an d lost of plasmid stability .To overcome this problem, the new synthetic glycerol minimal medium was optimized and formulated which enchanced leakage of phytase from periplasmic space to the extracellular medium. The extracellular phytase activity increased almost BB22 % approximately 1.8-fold in optimized medium when compared to un-optimized medium with the total phytase productivity o f 670j0 and 340.0 U L1 h r1, respectively after 10 hours of post-induction phase.The used of glycerol as the carbon source for the cell growth increased excretion of phytase outside the cell membrane and expression of the phytase is the highest among other types of carbon source.The yield coefficient of total phytase activity was in creased up to 102.92% which was 13,076.92 U g 'w hen statistically optimized induction strategy supplemented with glycine when compared with only lactose and CaCl3. Inconclusion, the increment of extracellular an d total phytase activity was not showing drastically improved but the productivity of the total phytase production and extra cellular phytase activity was increased up to 146.15 % and 119.9 %, respectively within 6 hours of post-induction phase

Production of erythromycin antibiotic by saccharoplyspora erythraea fermentation in shake flasks and bioreactor

Recently success of erythromycin in antibiotic market over the other antibiotics was due to that erythromycin has high quality and it is cheap in price. Erythromycin received much attention because of the increasing applications of its semi-synthetic modified derivatives to infection diseases, such as azithromycin, roxithromycin and clarithromycin. It is produced by the strain Saccharoplyspora erythraea (formerly known as Streptomyces erythraea). In this research, the aims were to optimize medium components for high erythromycin antibiotic production by the strain S. erythae via submerged fermentation using statistical technique known as response surface methodology. Glucose and yeast extract were found to have significant effect to erythromycin production using Placket-Burman experimental design for media screening. The Box-Benkhen experimental design was adopted for optimization studied. Finally, the optimal concentration of glucose, yeast extract, sodium nitrate, dipotasium hydrogen phosphate, sodium chloride and magnesium sulphate obtained using statistical media optimization is approximately 45;8; 4; 2.5;1.0; 0.5 (g L-1), respectively. Result showed that the maximal erythromycin concentration and CDW obtained in shake flasks of optimize medium were 412.5 mg L-1 and 4.9 g L-1, respectively. Production of erythromycin antibiotic reached 30.43% under the optimize medium. Furthermore, the batch culture using new medium formulation for erythromycin production was implemented using controlled and un-controlled pH conditions. Compared with the un-controlled pH bioreactor, the controlled bioreactor was increased erythromycin concentration by 12.9 % up to 567.5 mg L-1. This present work demonstrated that great potential production of erythromycin antibiotic at industrial scale

Optimization of exopolysaccharide production by pleurotus ostreatus using diffrent cultivation strategies

Pleurotus ostreatus or known as oyster mushroom was regarded as one of the most cultivated mushroom around the world. One of the qualities it has is it able to produced exopolysaccharide called pleuran which secreted into the medium during submerged fermentation. The polysaccharide composed mainly of ß-(1/3)-D glucose and ß-(1/6)-D glucose linked by glycosidic bond. It has molecular weight of 2.4 X 104 Da with molecular formula of (C6H10O5)x The importance of pleuran is that it has the immunomodulatory properties that associated in triggering our immune system response. Nowadays, submerged fermentation is considered as the best method in cultivation this kind of mushroom. However, the production process of this kind of mushroom and its exopolysaccharide production especially in term of medium component is still unclear. In this research, the objectives were to optimize the medium composition and to find the optimum carbon to nitrogen (C: N) ratio for high exopolysaccharide production. Eight different media was screened and followed by factor by factor optimization of the medium component. The factors that been studied were ideal concentration of glucose, yeast extract, ammonium sulfate and dipotassium phosphate. Media number six which contain glucose 60.0 g L-1, yeast extract 2.0 g L-1, (NH4)2SO4 5.0 g L-1, MgSO4.7H2O 0.2 g L-1, K2HPO4 1.0 g L-1 was selected as best media production for P. ostreatus cultivation . The experiment then was further with different concentration of each component in the medium six excluding magnesium sulfate heptahydrate which maintained at 0.2 g L-1 throughout all the experiment stage. The range concentration for glucose, yeast extract, ammonium sulfate and dipotassium phosphate was setup between 0 – 120 g L-1, 0 – 4 g L-1, 0 – 5 g L-1 and 0 – 2 g L-1 respectively. In order to get the best C: N ratio for highest exopolysaccharide production, eleven ratio of carbon to nitrogen was experimented ranging from 15:1 to 65:1.Result shown that the optimum concentration for glucose, yeast extract, ammonium sulfate and dipotassium phosphate was 80.0, 4.0, 2.5 and 1.0 g L-1 respectively whiles the optimal C: N ratio recorded was 40: 1. The optimized medium also produced 2.83 g L-1 of exopolysaccharide increasingly up to 49 % when compared with un-optimized medium which only produced 1.9 g L-1 of exopolysaccharide