Kim, Ryu-Ryun;Yi, Jeong-Hwan;Nam, Ki-Seok;Ko, Kyung-Won;Lee, Chan-Yong
Korean Journal of Microbiology
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v.47
no.1
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pp.14-21
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2011
Riboflavin synthase catalyzes the formation of one molecule of each riboflavin and 5-amino-6-ribitylamino-2,4-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrates, 6,7-dimetyl-8-ribityllumazine. The most remarkable feature is the sequence similarity between the N-terminal half (1-97) and the C-terminal half domain (99-213). To investigate the structure and fluorescent characteristics of the N-terminal half of riboflavin synthase (N-RS) in Escherichia coli, more than 10 mutant genes coding for the mutated N-terminal domain of riboflavin synthase were generated by polymerase chain reaction. The genes coding for the proteins were inserted into pQE vector designed for easy purification of protein by 6X-His tagging system, expressed, and the proteins were purified. Almost all mutated N-terminal domain of riboflavin synthases bind to 6,7-dimethyl-8-ribityllumazine and riboflavin as fluorescent ligands. However, N-RS C47D and N-RS ET66,67DQ mutant proteins show colorless, indicating that fluorescent ligands were dissociated during purification. In addition, most mutated proteins show low fluorescent intensity comparing to N-RS wild type, whereas N-RS C48S posses stronger fluorescent intensity than that of wild type protein. Based on this result, N-RS C48S can be used as the tool for high throughput screening system for searching for the compound with inhibitory effect for the riboflavin synthase.
1,4-Dioxane-degrading bacterial consortia were enriched from forest soil (FS) and activated sludge (AS) using a defined medium containing 1,4-dioxane as the sole carbon source. These two enrichments cultures appeared to have inducible tetrahydrofuran/dioxane and propane degradation enzymes. According to qPCR results on the 16S rRNA and soluble di-iron monooxygenase genes, the relative abundances of 1,4-dioxane-degrading bacteria to total bacteria in FS and AS were 29.4% and 57.8%, respectively. For FS, the cell growth yields (Y), maximum specific degradation rate ($V_{max}$), and half-saturation concentration ($K_m$) were 0.58 mg-protein/mg-dioxane, $0.037mg-dioxane/mg-protein{\cdot}h$, and 93.9 mg/l, respectively. For AS, Y, $V_{max}$, and $K_m$ were 0.34 mg-protein/mg-dioxane, $0.078mg-dioxane/mg-protein{\cdot}h$, and 181.3 mg/l, respectively. These kinetics data of FS and AS were similar to previously reported values. Based on bacterial community analysis on 16S rRNA gene sequences of the two enrichment cultures, the FS consortium was identified to contain 38.3% of Mycobacterium and 10.6% of Afipia, similar to previously reported literature. Meanwhile, 49.5% of the AS consortium belonged to the candidate division TM7, which has never been reported to be involved in 1,4-dioxane biodegradation. However, recent studies suggested that TM7 bacteria were associated with degradation of non-biodegradable and hazardous materials. Therefore, our results showed that previously unknown 1,4-dioxane-degrading bacteria might play an important role in enriched AS. Although the metabolic capability and ecophysiological significance of the predominant TM7 bacteria in AS enrichment culture remain unclear, our data reveal hidden characteristics of the TM7 phylum and provide a perspective for studying this previously uncultured phylotype.
Abdul Rahman, Nor Aini;Shirai, Yoshihito;Shimizu, Kazuyuki;Hassan, Mohd Ali
Biotechnology and Bioprocess Engineering:BBE
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v.7
no.5
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pp.281-288
/
2002
Recombinant Escherichia coli strain harboring the ${\lambda}$pR-pL promotor and heterologus poly-${\beta}$-hydroxybutyrate (PHB) biosynthesis genes was used to investigate the effect of culture conditions on the efficient PHB production. The expression of phb genes was induced by a temperature upshift from $33^{\circ}C\;to\;38^{\circ}C$. The protein expression levels were measured by using two-dimensional electrophoresis, and the enzyme activities were also measured to understand the effect of culture temperature, carbon sources, and the dissolved oxygen (DO) concentration on the metabolic regulations. AcetylCoA is an important branch point for PHB production. The decrease in DO concentration lowers the citrate synthase activity, thus limit the flux toward the TCA cycle, and increase the flux for PHB production. Since NADPH is required for PHB production, the PHB production does not continue leading the overproduction of acetate and lac-tate. Based on these observations, a new operation was considered where DO concentration was changed periodically, and it was verified its usefulness for the efficient PHB production by experiments.
Pen, B.;Iwama, T.;Ooi, M.;Saitoh, T.;Kida, K.;Iketaki, T.;Takahashi, J.;Hidari, H.
Asian-Australasian Journal of Animal Sciences
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v.19
no.9
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pp.1283-1290
/
2006
The effect of substituting potato by-products based silage (PBS) for concentrates on ruminal fermentation, methane production and nitrogen utilization in Holstein steers was evaluated. Three growing Holstein steers ($490{\pm}19kg$, initial body weight) were used in a $3{\times}3$ Latin square experiment with three experimental diets in which PBS was included at (1) 0, (2) 19 and (3) 27%, on a dry matter basis, replacing concentrates and wheat bran. Increasing replacement levels of PBS slightly increased dry matter (DM), organic matter (OM), NDF and ADF intakes by the steers (p<0.05). Inclusion of PBS at 19% increased crude protein (CP) digestibility of the experimental diets compared with the control (p<0.05). Substitution with PBS increased ADF digestibility and nitrogen retention (p<0.05), but did not affect energy retention. Energy loss as methane ranged between 5.0 and 6.1% of the total gross energy intake. There were no significant differences in carbon dioxide and methane production among all PBS levels, while daily methane production numerically increased with PBS inclusion. Substituting PBS for concentrates did not significantly affect ruminal pH and ammonia N concentration. Total VFA concentration, VFA molar proportions and blood metabolites were also unaffected by PBS replacement. These results suggest that substitution of PBS up to 27% of diet dry matter did not significantly increase methane production and was equal or superior to concentrates in ADF digestibility and nitrogen retention for growing steers.
Characteristics of organic matters (OM) in wastewater and the removal efficiencies were investigated using the influent and the effluent samples collected from 21 wastewater treatment plants. The OM characteristics investigated included biodegradability, humic content, specific UV absorbance (SUVA), the distribution percentage of refractory OM (R-OM), and synchronous fluorescence spectra. The types of wastewater (sewage, livestock waste/night soils, industrial waste) were easily distinguished by comparing the synchronous fluorescence spectra of the influent wastewater. The prominent peak of protein-like fluorescence (PLF) was observed for livestock waste/night soils whereas sewage exhibited a unique fluorescence peak at a wavelength of 370 nm. Irrespective of the wastewater types, the distribution percentage of R-OM increased from the influent to the effluent. Livestock waste/night soils showed the highest removal efficiency among all the three types of wastewater. There was no statistical difference of the removal efficiency between a traditional activated sludge and biological advanced treatment processes. Removal efficiency based on dissolved organic carbon DOC presented good correlations with the distribution percentage of R-OM and fulvic-like fluorescence (FLF) of the influent. The prediction for DOC removal efficiency was improved by using multiple regression analyses based on some selected OM characteristics and mixed liquid suspended solid (MLSS).
In the present study, a yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp. VITGBN2 based on molecular techniques and was found to be a potent producer of acidic diacetate sophorolipid in mineral salt media containing vegetable oil as additional carbon source. The chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. This sophorolipid was used as a stabilizer for synthesis of zinc oxide nanoparticles (ZON). The formation of biofunctionalized ZON was characterized using UV-visible spectroscopy, XRD, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy. The antimicrobial activities of naked ZON and sophorolipid functionalized ZON were tested based on the diameter of inhibition zone in agar well diffusion assay, microbial growth rate determination, protein leakage analysis, and lactate dehydrogenase assay. Bacterial pathogen Salmonella enterica and fungal pathogen Candida albicans showed more sensitivity to sophorolipid biofunctionalized ZON compared with naked ZON. Among the two pathogens, S. enterica showed higher sensitivity towards sophorolipid biofunctionalized ZON. SEM analysis showed that cell damage occurred through cell elongation in the case of S. enterica, whereas cell rupture was found to occur predominantly in the case of C. albicans. This is the first report on the dual role of yeast-mediated sophorolipid used as a biostabilizer for ZON synthesis as well as a novel functionalizing agent showing antimicrobial property.
Our government has performed to support the nation-wide application of customized fertilizer based on soil-testing results and crop nutrient balance in order to promote the environment-friendly agriculture and to respond the global environment guide-line since 2010. This study was performed at the selected local paddy fields (Hwaseong-si, Uiseong-gun and Miryang-si) with different soil chemical properties in 2012. The contents of amino acids measured showed an increasing trend with fertilization, and glutamic acid was the most abundant amino acid followed by aspartic acid, leucine and alanine. However, valine, isoleusine, tyrosine and lysine were not significantly affected by fertilization. The significant differences in grain N, expressed as a crude protein, and amino acids dose was observed between experimental sites (p<0.001), treatments (p<0.01 to 0.001) and interaction of both factors (p<0.01 to 0.001). In our experiment the following order of carbon skeleton backbones to produce amino acids was observed irrespective of experiment sites and fertilization: ${\alpha}$-ketoglutarate > oxalate > pyruvate > 3-phosphoglycerate > phosphoenolpyruvate. In conclusion, customized fertilizer had no difference in amino acids compared to the conventional-NPK practice which was higher than in no fertilization, and also the normal paddy represented slightly higher amino acids compared to the reclaimed. Further study based on the present results is required to investigate what is main factor to amino acids between genetic and environmental factors.
The processing conditions and quality of sardine surimi were examined: Raw sardine meat was separated, washed in 0.2% $NaHCO_3$ and 0.15% NaCl solution, and then dewatered by centrifuge. The dewatered sardine meat was chopped, mixed with 20% emulsion curd (soybean protein : water : refined sardine oil=1:5:2.6), 4% sorbitol, 4% sucrose, 0.2% polyphosphate and 0.1% sodium erythorbate by stone mortar. The mixed sardine meat was frozen with contact freezer, packed in carton box and then stored at $-25{\pm}2^{\circ}C$. The moisture, crude protein and lipid contents of the sardine surimi product was 73.3%, 15.0% and 6.9%, respectively. Fatty acid composition of product consisted of 28.8% of saturates, 24.3% of monoenes and 47.7% of polyenes and the major fatty acids were 16:0, 20:5, 18:1, 22:6 and 16:1. The results of changes in POV, TBA value, fatty acids, texture and sensory score of products during frozen storage showed that lipid oxidation and freeze denaturation of product could be retarded, and flavor enhanced by addition 20% emulsion curd and 0.1% sodium erythorbate. In an attempt to apply sardine surimi in producing surimi-based product, it was concluded that pollack surimi could be substituted with sardine surimi up to 40% without showing any significant changes in texture and taste of surimi-based product.
Young-kyu Lee;Yeong Seo Choi;Myoung cheol Moon;Jae min So;Ohkyung Kwon;Wonsil Choi;Joon weon Choi;In Yang
Korean Chemical Engineering Research
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v.62
no.1
/
pp.87-98
/
2024
This study was conducted to investigate the efficiency of the filtering sets composed of fiberboards, which were fabricated with lignocellulosic fiber and cork oak bark-based activated carbon (COA), as well as traditional Korean paper handmade from mulberry trees (KP) for the filtration of PM, TVOC and HCHO. Three-layers fiberboards (WRF) were fabricated with wood fiber in its surface layers and recycled fiber/COA in its core layer using a protein-based adhesive with the resin content of 8%. Filtering sets were composed of three WRF and one sheet of KP. Concentrations of PM, TVOC and HCHO generated with the combustion of a incense in a sealed laboratory hood were reduced efficiently with the operation of air-purifier installed the filtering sets. Except for the WRF fabricated with 4%/4% resin contents, other WRF were prepared with 5%/3% and 6%/2% resin contents in surface/core layers, and then the WRF were used with KP for the fabrication of filtering sets. Filtration efficiency of the filtering sets was improved as the core-layer resin content applied in the fabrication of WRF decreased. In addition, filtration efficiency of the WRF-based filtering set fabricated with KP of 25 g/m2 basis weight was higher than that with KP of 45 g/m2 basis weight. Filtering sets composed of three-layers fiberboards (RWF) that recycled fiber and wood fiber/COA were used in its surface and core layers, respectively, and KP-25g showed higher filtration efficiency than those of WRF-based filtering sets. Air-inhalation equipment installed the RWF-based, WRF-based filtering sets and without filtering set were operated in small indoor and large outdoor spaces. Efficiency for filtering PM and TVOC of the RWF-based filtering sets was higher than that of other filtering sets. It is concluded that fiberboard-based filtering sets composed of RWF and KP-25g can be used as a filter for reducing the concentrations of PM and TVOC existed in indoor and outdoor spaces.
A repeated $4{\times}4$ Latin square design was conducted with eight ileal cannulated castrates to examine the effect of source of starch and fiber on nutrient balance and energy metabolism. Pigs were fed on one of the four experimental diets: Control diet (C) mainly based on cooked rice; and diets P, S and W with the inclusion of either raw potato starch, sugar beet pulp or wheat bran supplementation, respectively. With the exception of an increased (p<0.05) energy loss from methane production with diet S observed, no significant differences (p>0.05) in the ratio of metabolizable energy (ME)/digestible energy, the utilization of ME for fat deposition and for protein deposition, energy loss as hydrogen and urinary energy were found between diets. The efficiency of utilization of ME for maintenance was lower (p<0.05) with diets P and S than with diet C. The inclusion of fiber sources (sugar beet pulp or wheat bran) or potato starch reduced the maintenance energy requirement. The fecal energy excretion was increased (p<0.05) with either sugar beet pulp or wheat bran supplementation, while it was unaffected (p>0.05) by addition of potato starch. In comparison with diets C and P, a lowered ileal or fecal digestibility of energy with diets S and W was observed (p<0.05). Feeding sugar beet pulp caused increased (p<0.05) daily production of methane and carbon dioxide and consequently increased energy losses from methane and carbon dioxide production, while it did not influence the daily hydrogen production (p>0.05). An increased (p<0.05) proportion of NSP excreted in feces was seen by the supplementation of wheat bran. Higher NSP intake caused an increased daily amount of NSP in the ileum, but the ileal NSP proportion as a percentage of NSP intake was unaffected by diets. Feeding potato starch resulted in increased daily amount of starch measured in the ileum and the proportion of ileal starch as a percentage of starch intake, while no significant influence on fecal starch was found. Higher (p<0.05) daily amount of fecal starch and the proportion of fecal starch as a percentage of starch intake were found with fiber sources supplementation compared with diets C and P. By increasing the dietary NSP content the fecal amount of starch increased (p<0.01).
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