β-Glucan is one of the most abundant polysaccharides in yeast Saccharomyces cerevisiae cell wall. The aim of this research is to explore an alternative nitrogen sources for β-glucan production. S. cerevisiae were grown in fermentation medium with different nitrogen sources. Peptone 2%, glutamic acid 0,5%, urea 0,2%, and diammonium hydrogen phosphate (DAHP) 0,02% were used for nitrogen source in the medium. A two liter air-lift fermentor was used in the fermentation process for 84 hours (T = 300C, pH 7, and 1.5 vvm for the aeration). During the fermentation, optical density, extraction of β-glucan, glucose and protein in hydrolisate cultured were determined. β-glucan production level is similar with the growth rate of yeast and followed by decreasing glucose and protein content in hydrolysis cultured. The highest and lowest β-glucan content were obtained from peptone (933.33 mg/L) and glutamic acid (633.33 mg/L) as a nitrogen source in cells cultured after fermentation completed respectively. Yeast cells cultured with urea and DAHP as a nitrogen source give the same content of β-glucan about 733.33 mg/L. β-glucan concentration produced in medium with urea was a higher than that produced usingglutamic acid and DAHP as a nitrogen source. The result indicated that urea can be used as an alternative nitrogen source for theproduction of β-glucan. Urea is easily available and cheaper than peptone, glutamic acid and DAHP.© 2007 Jurusan Biologi FMIPA UNS SurakartaKey words: β-glucan, Saccharomyces cerevisiae, air-lift fermentor
Indonesia is a rich biodiversity country where various medicinal plants have existed. One species of medicinal plants is Globe Amaranth (Gomphrena globosa, Amaranthaceae). This species is native to Central America and has been widely spread to the tropics. To date, the species can be easily found in the home gardens as an ornamental plant. Medicinal plants have been used for generations by traditional people. It was empirically proven that medicinal plants have the ability to cure certain diseases such as dysentery. All parts of this plant can be used as medicine. However, only the flower of the species was used in this study. The objective of the study was to identify the highest antimicrobial activity of Gomphrena globosa flower extract using ethanol, petroleum ether, ethyl acetate and n-butanol solvents. Gomphrena globosa flower was extracted using 96% ethanol and then was by partitioned using petroleum ether, ethyl acetate, and n-butanol respectively. The extracts were then evaporated using a rotapavor until condensed extract was obtained. Phytochemical screening was done on both of the flower powder and extract. The result of Pharmacognosy evaluation of the Globe Amaranth flower as follows: water content 8.17%, total ash content 9.11%, acid-insoluble ash 1.50%, acid-soluble ash 6.43%, watersoluble extract 10.79%, ethanol-soluble extract 3.51% and dry content 10.19%. The condensed extracts were tested for antimicrobial activity against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Shigella dysenteriae. Result of antibacterial activity test by agar diffusion method showed that the higher concentration of the extract led to higher bacterial inhibition zone. The highest antimicrobial activity was obtained from n-butanol extract as indicated by a significant inhibition zone around the paper disk.
Production of β-glucan by Agrobacterium sp is influenced by the composition of nutrition in the fermentation media. Molases has been used successfully by others in the fermentation media of S. cerevisiae to increase the yield of-glucan, and similarly, uracil has been used in the fermentation media of Agrobacterium sp to increase the yield of-glucan. Investigations to increase the yield of-glucan by two strains of Agrobacterium sp, i.e. A1.5 (reference) and B4.4 (local strain), have been carried out by addition of various combination of molases and uracil into fermentation media, i.e. 5%(v/v) molase-0,05%(b/v) uracil; 5% molase-0,025% uracil; 10% molase-0,05% uracil; and 10% molase-0,025% uracil. The β-1,3-glucan and β-1,2-glucan fractions were separated by extraction method. Beta-glucan concentration was determined as the glucose monomer using the phenol-sulphate spectrophotometric method at 490 nm. The protein content was determined by a modified Lowry-spectrophotometric method at 750 nm. The results showed that all combination of molases and uracil in the fermentation media of Agrobacterium sp A1.5 and B4.4 strains have increased both the dry-weight yield of β-glucan (crude) and the βglucan content, with the highest was in a medium containing 10% molases-0,025% uracil combination. In the above medium, the A1.5 strain produced the highest β-glucan (7,5%) with the lowest protein content (8,4%) in the β-1,3-glucan fraction, while the β-glucan content in the β-1,2-glucan fraction were all lower than in the control media, while the protein content were all higher than in the control media. In the above media, the B4.4 strain produced the highest β-glucan, 7,2% in the β-1,3-glucan fraction, and 13,1% in β-1,2-glucan fraction, while the lowest protein content (8,4%) was in the β-1,3-glucan fraction. In conclusion, fermentation media of Agrobacterium sp A1.5 strain or B4.4 strain containing molase and uracil combination have increased both the dry-weight yield of total β-glucan (crude) and the β-glucan content, while reduced the protein content. There is no clear FTIR spectrum difference between supposedly β-1,2-glucan fraction and β-1,3-glucan fraction.
ABSTRAKThe need of â-glucan is increasing in food, medicine and cosmetic industry, because it becomes anticancer, antitumor and antiaging, increases immunosystem, and decreases cholesterol content in blood. The cell walls of S. cerevisiae contain 80-90% polysaccharides that posses â-glucan. This research was aimed to obtain appropriate carbon sources to increase the production of â-glucan. The carbon sources used were glucose, glucose commercial, sucrose and molases. The fermentation process was done by using air lift fermentor. The steps of fermentaton included regeneration of S. cerevisiae strain, preculture, fermentor preparation and running fermentor for 84 hours. Sampling of S. cerevisiae culture was determined the cell growth by optical density (OD) using spectrophotometer UV/VIS at ë 550 nm. The protein content was determined by Lowry method at ë 755 nm and the total glucose was measured by phenol sulphate method at ë 490 nm. The measurement result of cell growth showed that the high intensity of S. cerevisiae in medium contain molases, but it did not show significant effect when compare to other carbon sources. The protein and carbohydrate contain in medium tended to decrease. The result of â-glucan on glucose, sucrose, glucose commercial and molases were 933,3, 1100, 1000, and 966,7 mg/l. It can be concluded that sucrose and glucose commercial can replace the glucose to produce of âglucan, because they are cheaper and easier to get. Beside that, molases can be used as an alternative carbon source because it can produce of â-glucan as well as glucose.
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