Do, Thi Huyen;Dao, Trong Khoa;Nguyen, Khanh Hoang Viet;Le, Ngoc Giang;Nguyen, Thi Mai Phuong;Le, Tung Lam;Phung, Thu Nguyet;Straalen, Nico M. van;Roelofs, Dick;Truong, Nam Hai
Asian-Australasian Journal of Animal Sciences
/
v.31
no.5
/
pp.738-747
/
2018
Objective: In a previous study, analysis of Illumina sequenced metagenomic DNA data of bacteria in Vietnamese goats' rumen showed a high diversity of putative lignocellulolytic genes. In this study, taxonomy speculation of microbial community and lignocellulolytic bacteria population in the rumen was conducted to elucidate a role of bacterial structure for effective degradation of plant materials. Methods: The metagenomic data had been subjected into Basic Local Alignment Search Tool (BLASTX) algorithm and the National Center for Biotechnology Information non-redundant sequence database. Here the BLASTX hits were further processed by the Metagenome Analyzer program to statistically analyze the abundance of taxa. Results: Microbial community in the rumen is defined by dominance of Bacteroidetes compared to Firmicutes. The ratio of Firmicutes versus Bacteroidetes was 0.36:1. An abundance of Synergistetes was uniquely identified in the goat microbiome may be formed by host genotype. With regard to bacterial lignocellulose degraders, the ratio of lignocellulolytic genes affiliated with Firmicutes compared to the genes linked to Bacteroidetes was 0.11:1, in which the genes encoding putative hemicellulases, carbohydrate esterases, polysaccharide lyases originated from Bacteroidetes were 14 to 20 times higher than from Firmicutes. Firmicutes seem to possess more cellulose hydrolysis capacity showing a Firmicutes/Bacteroidetes ratio of 0.35:1. Analysis of lignocellulolytic potential degraders shows that four species belonged to Bacteroidetes phylum, while two species belonged to Firmicutes phylum harbouring at least 12 different catalytic domains for all lignocellulose pretreatment, cellulose, as well as hemicellulose saccharification. Conclusion: Based on these findings, we speculate that increasing the members of Bacteroidetes to keep a low ratio of Firmicutes versus Bacteroidetes in goat rumen has resulted most likely in an increased lignocellulose digestion.
Many factors exert an influence on enzyme activity and thus on the rate of reactions that they catalyse. The most important of these factors are pH, temperature, substrate concentration, and the concentration of some inhibitors present. A solution of the enzyme diastase, which breaks down molecules of the polysaccharide starch to the disaccharide maltose by hydrolysis, was provided. Activity of this enzyme was measured by the rate at which starch was removed from the reaction mixture. These experiments were designed to study this reaction rate under varying conditions and the following results were obtained. 1. The range of optimum pH for this enzyme at room temperature was 4.0-7.0 and the optimum pH was 5.0. 2. The range of optimum temperatures for this enzyme at pH 7.0 was $30^{\circ}C-50^{\circ}C$ and the optimum temperature was $40^{\circ}C$. 3. The relationship between the enzyme activity and substrate concentration could be expressed by the Michaelis-Menten equation. The limiting velocity of this enzyme at room temperature and pH 7.0 was $415{\mu}g$ starch removed/ml of reaction mixture/min and $K_m$, Michaelis constant, was $343{\mu}g/ml$. 4. Inhibitors NaCl and $HgCl_2$ blocked this enzyme activity completely at 1% and 0.01% respectively.
Journal of the Korean Society of Food Science and Nutrition
/
v.10
no.1
/
pp.85-91
/
1981
The growing cells of S. aureus were fractionated along the Schmidt-Thannhauser-Schneider's technique into several fractions such as TCA(trichloroacetic acid)-soluble, lipid, nucleic acid, protein and residue fraction. They were also fractionated according to their cellular structure into Sonic-supernatant, SDS(sodium lauryl sulfate)-soluble, Formamide-soluble and Residue fraction. Fractionation was carried out by orderly treatment of the Sonic pellet with 1.0% SDS and hot$(150^{\circ}C)$ formamide, and the pellet was prepared by centrifugation of the cells sonic osillated for 20 minutes at 150 watt. Sonic-supernatant fraction contained a 91.3% of total DNA while other fractions contained less than 9.5%. SDS-soluble fraction showed a high activity of malate dehydrogenase(13.67 unit/mg protein) and which was higher 22.3 times than the activity found from unsoluble fraction. Formamide-soluble fraction prepared from SDS-undoluble pellet by using the hot formamide exhibited a clear action of reducing sugars against the Anthronesulfate, while it exhibited no clear action against the ninhydrin. However, contrastly, the residue remainnning after extraction with formamide exhibited a clear action against ninhydrin and glucosamine was detected form the hydrolysate of residue by paper chromatography. From these results it is considered that the Sonic-supernatant fraction is mainly consisted of plasmic component of the cells. Other fractions, SDS-soluble, Formamide-soluble and Residue, should be consisted of plasma membrane, lipoplysaccharide and peptidoglycan of the cell, respectively.
Chitosan is abundant polysaccharide polymer in nature Chitosan has been the subject of much research regarding its potential as a nutritional adjunct and pharmaceutical ingredient. In the present study, we examined fracture-healing process with chitosan administration and low calcium diet in rats. Left femur was fractured and fixed with intramedullary pin. The rats were fed normal diet or low calcium diet and administered chitosan with the doses of 0, 50, 100 and 150ng/ml orally 5 times a week for 10 weeks. Soft X-ray and mechanical testing of all fractured femora was taken. Radiographical finding showed that the callus formation and process of fracture healing was present in all the fractured femora. Mechanical testing indicated that the maximum load and stiffness of femur in rat fed low calcium diet was lower than those of that in rat fed normal diet. No difference in maximum load and stiffness of fractured femora in chitosan treated rat were observed as compared to vehicle treated rat. Chitosan or/and low calcium diet did not affect the ratio of fractured/unfractured femur about maximum load and stiffness. The results suggest that chitosan dose not affect the bone mechanical strength and the process of fracture healing. Low calcium diet does decrease the bone mechanical strength.
The rationale for the use of fungi in treating waste streams from food processing plants I~as been that of incorporating the dissolved and suspending nutrients into a macroscopic organism which can be filtered out readily. In order for a process using fungi to meet these objectives we examined a strain of fungi, Aspergillus fumigatus, which grew well on a variety of polysaccharide-containing materials and showed both efficient BOD removal and high quality protein recovery. In this experiment the fungal choice was based on the laboratory screening studies where the criteria used was BOD and COD reduction, growth response, mycelial yield, and the ability to compete with the natural flora. In the fermentation system used far the continuous culture of Aspergillus fumigatus the best combination of operating variables, inoculum ratio, temperature, initial pH, fermentation time and agitation rate was 5%(v/v), $35{\sim}40^{circ}C,\;pH\;4.5{\sim}5.0$, 2days and 150rpm, respectively. The fungus reduced BOD and COD to 94.0 and 90.4%, respectively and 3.15g of dry mycelium per liter of alcohol waste was harvested during 48hr of incubation time. The protein efficiency ratios for the control diet and the experimental diet containing the fungal protein were $3.42{\pm}0.15$ and $3.40{\pm}0.43$, respectively.
Park, Ki-Nam;Song, Hyun-Chul;Jee, Yu-Jin;Yoo, Jin-Young
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.31
no.2
/
pp.116-129
/
2005
Distraction osteogenesis is a new bone formation technique. There is a advantage of the environmental adaptation when distraction force is applied to the gap between osteotomy lines. But it has a disadvantage of long-term wearing of the appliance and long consolidation period. Therefore we make an effort to reduce it and repair normal function. Extracellular matrix proteins have a function to control the cellular growth, migration, shape and metabolism. In these, hyaluronic acid is a member of polysaccharide glycosaminoglycans (GAGs) and has a important function as bone formation and osteoinduction property. Purpose : In this experimental study in rabbit mandibular distraction osteogenesis, we investigated the bone enhancing property of hyaluronic acid and the expression of extracellular proteins such as osteocalcin and osteonectin. Materials and Methods : The experimental study was carried out on 24 Korean male white rabbits (both mandibular body, n=48). Distraction group was divided to distraction experimental (A, n=16) and distraction control (B, n=16) by the application of hyaluronic acid (Hyruan, LGCI, Seoul, Korea). Normal control group (C, n=16) was only osteotomized. After 5 days latency, distraction devices were activated at a rate of 1.4 mm per day (0.7 mm every 12hours) for 3.5 days. Animals were sacrificed at postoperative 3, 7, 14, and 28 days. H&E stain and immunohistochemical stain was done on decalcified section. Additionally RT-PCR analysis was done for the identification of the expression of osteocalcin and osteonectin. Results : The bone formation in distraction experimental group was much more than that in distraction and normal control group at postoperative 28 days. In immunohistochemical stain, osteocalcin was enhanced at only postoperative 14 days, but osteonectin was not different at each post-operation days. In RT-PCR analysis, osteocalcin was not different at each post-operation days, but osteonectin was strongly expressed in distraction experimental group at postoperative 7 days. The expression of osteocalcin and osteonectin was elevated during the healing period. Conclusion : We found the good bone formation ability of hyaluronic acid in distraction osteogenesis through the immunohistochemistry and RTPCR analysis to osteocalcin and osteonectin, known as a bone formation marker. The application of hyaluronic acid in distraction osteogenesis is a method to reduce the consolidation period.
Marine bacterium Bacillus cereus ASK202 produced an extracellular agarase (E.C.3.2.1.81) which showed a high level of enzyme activity in the presence of agar and agarose. In the optimal culture conditions, the agarase production increased 7.7 folds compared with the one obtained from the basal medium. Agarase production reached upto 160 units/L after 24hr of cultivation in a modified marine medium at $25^{\circ}C$. The degree of purification increased 31.5 folds with 27.8% yield through freeze drying, DEAE Sepharose CL-6B and Superose 6HR 10/30 column chromatography. The molecular weight of the purified agarase was determined to be 90,000 daltons by gel-permeation filteration. Optimal temperature and pH for the enzyme activity were $40^{\circ}C$ and 7.8, respectively. The enzyme was stable up to $50^{\circ}C$ and at a broad pH range of 5.0-10.0. The $\beta$-agarase was activated by $Zn(NO_3)_2$, and was inhibited by $CuSO_4$ and $SnCl_2$. The Km and Vmax values of this enzyme for agarose as a substrate was $2.4mg/m\ell$ and 13.6 mg/m$\ell$, respectively.
In order to use a polysaccharide, curdlan, as a concrete admixture, we first developed a pilot-scale fermentation process for the mass production of curdlan. We also examined the rheological properties of curdlan, and tested how well the curdlan obtained in this work increased the segregational resistance of the cement slurry. Fermentation was performed in a 300-liter fermenter equipped with 3 disk-turbine impellers. Since curdlan production is stimulated under nitrogen-limiting conditions, the culture pH was shifted from the optimal pH for cell growth (pH 7.0) to the optimal pH for curdlan production (pH 5.5) at the onset of ammonium exhaustion. We obtained a curdlan production of 65 g/L in 120 hr batch cultivation of Agrobacterium species. The insoluble curdlan at the final stage of fermentation was readily harvested by centrifugation together with the cells. The freeze-dried sample contained 78% (w/w) of curdlan. The solubility and viscosity of the curdlan increased with the increase of the solution pH, which enhances the viscosity of concrete since the pH of concrete is extremely high (pH 13.0). Test results of the curdlan as a concrete admixture with cement slurry demonstrated that it prohibits the leakage of water. In conclusion, this work certifies and enlarges curdlan's industrial potential as a concrete admixture.
Journal of the Korean Society of Food Science and Nutrition
/
v.33
no.2
/
pp.278-286
/
2004
The objective of the current study was to determine the effects of arabinoxylane and PSP on mouse splenocytes, T cells, B cells and macrophages in vitro. Arabinoxylane and PSP directly induced the proliferation of spleen cells in a dose-dependent manner and increased IFN-${\gamma}$ synthesis. Especially, PSP induced IL-2, IL-4 and IL-10 production, Both arabinoxylane and PSP increased PFC (plaque forming cell) and RFC (rosette forming cell) formation. Arabinoxylane was not induced the proliferation of T cells, but PSP directly induced the proliferation of T cells in a high dose. Arabinoxylane and PSP increased the proliferation of B cells and the phagocytic effects of macrophage. When arabinoxylane and PSP were used in macrophage cell line stimulation, there was a marked induction of NO synthesis in a dose-dependent and an increased TNF-$\alpha$ and IL-6 synthesis. Especially, PSP also induced IL-1$\beta$ production. When arabinoxylane and PSP treated in macrophage cell line, there was induction of MHC class II expression. These results suggest that the capacity of arabinoxylane andPSP seem to act as a potent immunomodulator causing augmentation of immune cell activity, and with the absence of notable side-effects, arabinoxylane and PSP could be used as a biological response modifier having possible therapeutic effects against immunological disorders.
Journal of the Korean Society of Food Science and Nutrition
/
v.44
no.2
/
pp.191-199
/
2015
To elucidate the immuno-stimulatory activity of traditional fermented vinegar, six kinds of crude polysaccharides were isolated from traditional fermented vinegars manufactured with different raw materials in domestic or foreign countries, after which their chemical properties and immuno-stimulatory activities were evaluated. Of the six samples, three kinds of crude polysaccharides prepared from Korean brown rice vinegar (KBV-0), Japanese brown rice vinegar (JBV-0), and Korean persimmon vinegar (KPV-0) showed higher yields and interleukin (IL)-6 production by macrophages and were thus selected for further study. Anti-complementary activities of KBV-0, KPV-0, and JBV-0 increased dose-dependently. KBV-0 and KPV-0 showed higher anti-complementary activities ($ITCH_{50}$ 62 and 65%) than JBV-0 at $1,000{\mu}g/mL$. KBV-0, KPV-0, and JBV-0 did not affect growth of peritoneal macrophages at a dose of 1.6 to $1,000{\mu}g/mL$, where as they significantly augmented production of IL-6, IL-12, and TNF-${\alpha}$ in a dose-dependent manner. However, immuno-stimulatory activity of KPV-0 was the most potent among the tested polysaccharides. These results suggest that Korean fermented vinegars contain selected polysaccharides that confer immuno-stimulatory activities beneficial to human health.
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