This study was conducted to assess the relation between threonine (Thr) oxidation rate and threonine efficiency on rat and chicken fed with graded levels of protein and threonine. The increase in threonine content from 0.28 to 0.72% in a diet containing 12.0% crude protein (CP) caused a gradual increase in threonine dehydrogenase (TDG) activity in rat liver. Similar, but more pronounced results were observed after 18.0% CP in the diet. Both protein levels in combination with the highest level of threonine supplementation increased liver TDG activity significantly, indicating enhanced threonine catabolism. Parameters of efficiency of threonine utilization calculated from parallel nitrogen balance studies decreased significantly and indicated threonine oversupply after a maximum of threonine supplementation. At the lower levels of threonine addition the efficiency of threonine utilization was not significantly changed. In the chicken liver up to 0.60% true digestible threonine (dThr) in the 18.5% CP diet produced no effect on the TDG activity. However, TDG activity in the liver was elevated by the diet containing 22.5% CP (0.60% dThr) and the efficiency of threonine utilization decreased, indicating the end of threonine limiting range. In conclusion, the in vitro TDG activity in the liver of rat and growing chicken has an indicator function for the dietary supply of threonine.
The aim of this investigation was to determine the influence of different protein and threonine (Thr) levels on the liver threonine dehydroaenase (TDG) activity b\ulcorner rats. In rats fed on CP (crude Protein) - diets, TDG activity was increased during an CP rise to 12.0% CP, decreased slightly down to 18.0% CP and showed .a trend to increase from 18.0 to 24.0% CP. In rats the feeding with graded Protein supply gavee no indication for additional stimulation of threonine-oxidation by TDG over a wide range of CP-content in the diets. The increase in threonine content from 0.28 to 0.72% in the presence of 12.0% CP caused a gradual increase in TDG activity in rat liver. This similarly applied to the feed admixed with 18.0% CP, but at a higher level.
This study was carried out to evaluate the relationship between threonine (Thr) efficiency and Thr dehydrogenase (TDG) activity as an indicator of Thr oxidation on chicks fed with levels of diets (CP [17.5% and 21.5%] and Thr [3.8 and 4.7 g/100 g CP]; glycine [Gly][0.64% and 0.98%] and true digestible Thr [dThr] [0.45% and 0.60%]). Calculation of the Thr efficiency was based on N-balance data and an exponential N-utilization model, and TDG activity was determined as accumulation of aminoacetone and Gly during incubation of hepatic mitochondria. This study found that in the liver of chicks who received a diet containing up to 0.79% Thr (4.7 g Thr/100 g of CP) in the 17.5% CP diet, no significant (p>0.05) effect on TDG activity was observed. However, significantly (p = 0.014) increased TDG activity was observed with a diet containing 21.5% CP (4.7 g Thr/100 g of CP) and the efficiency of Thr utilization showed a significant (p = 0.001) decrease, indicating the end of the Thr limiting range. No significant (p>0.05) effect on the total TDG activity and accumulation of Gly was observed with addition of Gly to a diet containing 0.45% dThr. In addition, addition of Gly to a diet containing 0.60% dThr also did not result in a change in accumulation of Gly. Due to an increase in accumulation of aminoacetone, an elevated effect on total TDG activity was also observed. No significant (p>0.05) reduction in the efficiency of Thr utilization was observed after addition of Gly at the level of 0.45% dThr. However, significantly (p<0.001) reduced efficiency of Thr utilization was observed after addition of Gly at the level of 0.60% dThr. Collectively, we found that TDG was stimulated not only by addition of Thr and protein to the diet, but also by addition of Gly, and efficiency of Thr utilization was favorably affected by addition of Gly at the level near to the optimal Thr concentration. In addition, no metabolic requirement of Gly through the TDG pathway was observed with almost the same accumulation of Gly and a slight increase in TDG activity by addition of Gly. Thus, our findings suggest that determination of TDG activity and parameter of efficiency of Thr utilization may be useful for evaluation of dietary Thr level.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.20
no.1
s.32
/
pp.1-15
/
2007
Objectives : RVC has long been used for a useful natural agent ameliorating inflammation related symptoms in the folk medicine recipe. This study was performed to investigate effects of RVC on the inflammation and oxidation in RAW 264.7 cells. Methods : The RVC was extracted with 80% ethanol and sequentially partitioned with solvents in order to increase polarity. With the various fractions, we determined the activities on the inflammation and oxidation in RAW 264.7 cells. Results : 1. Among the various solvent extracts of RVC, the butanol fraction showed the most powerful inhibitory ability against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells without affecting cell viability. 2. Butanol fraction showed a oxidation inhibition effect by decreasing the DPPH and OH radicals. 3. Butanol fraction exhibited the inhibitory avilities against iNOS and COX-2. 4. Reverse transcriptase polymerase chain reaction (RT-PCR) and Westem blotting analysis revealed that the BuOH fraction provided a primary inhibitor of the iNOS protein and mRNA expression in LPS-induced RAW 264.7 cells. Among the up-regulater molecules of iNOS and COX-2, the BuOh fraction of RVC was shown the inhibitory activity of phoshporylation of c-Jun N-terminal kinase (JNK) 1/2 and threonine protein kinase (AKT), the one of the MAPKs pathway. Conclusion : Thus, the present study suggests that the response of a component of the BuOH fraction to NO generation via iNOS expression provide a important clue to elucidate anti-inflammatory and anti-oxidation mechanism of RVC.
This study was designed to describe the effect of the protein quality at different intake level of protein on the protein metabolism in the whole body of growing pigs with a simulation model. Varying to the protein level in feeds, four simulations were conducted. The feed protein level, represented as proportions of digestible protein to the metabolic energy (DP/ME, g/MJ), were 6-8, 11-13, 17-19, and 23-25 DP/ME, respectively. Two protein quality and six weeks of growth time were used at each simulation. The objective function for the simulations was protein deposition in the whole body, which was calculated from the experimental results. The parameters in the simulation were determined by the parameter estimation technique. The results obtained from the simulation were as follows: The protein synthesis and breakdown rates(g/day) in the whole body was increased with the increase of protein quality only at lower or required level of protein intake. They showed a parallel behavior in the course of growth, irrespective of quality and level of feed protein intake. The simulated protein deposition and protein synthesis showed a linear relationship between them at different protein quality and level. The affinity parameter showed a linear relationship between them at different protein quality and level. The affinity parameter showed that arginine, tryptophan and isoleucine were more efficient in the stimulation ofbody protein synthesis. Lysine and phenylalanine+tyrosine were less efficient. The oxidation parameter showed that histidine, pheyalanine+tyrosine were less efficient. The oxidation parameter showed that histidine, phenyalanine+tyrosine, and methionine+cystine were oxidized in larger magnitude than lysine and threonine. The oxidation parameter of most amino acids increased with the increase of protein intake beyond the requirement level, but not any more at highest protein intake level. Finally it was found that the improvement of feed protein quality at the lower or required level of protein intake increase protein deposition through a parallel increase of protein synthesis and breakdown.
Oriental pear (Pyrus communis L. cv 'Niitaka') was stored at $0^{\circ}C$ for 5 months and major metabolites involved in blackening of the peel were analyzed by untargeted GC-MS and targeted HPLC methods. In this study, peels of sound and skin-blackened pears were analyzed and compared. Skin-blackened fruit was clearly characterized by a distinctive pattern in changes which included a decrease of malic acid, succinic acid, and ascorbic acid, while an increase of fumaric acid, threonine, and gluconic acid, which indicated both reduced metabolic activity and anti-oxidative capacity of the cells. Chlorogenic acid was a major phenolic compound and the peel of sound fruit showed high levels of free phenolic compounds compared than the peel of skin-blackened fruit which are believed to be related to oxidation of phenolics in skin-blackened tissue. The changes or profiling of major metabolites by targeted or untargeted analysis method could become a useful tool for understanding physiology, disorder mechanism, and identifying metabolic networks connecting primary and secondary metabolism in postharvest research.
Seok, Yun-Jeong;Her, Jae-Young;Kim, Yong-Gun;Kim, Min Yeop;Jeong, Soo Young;Kim, Mina K.;Lee, Jee-yeon;Kim, Cho-il;Yoon, Hae-Jung;Lee, Kwang-Geun
Toxicological Research
/
v.31
no.3
/
pp.241-253
/
2015
Furan ($C_4H_4O$) is a volatile compound formed mostly during the thermal processing of foods. The toxicity of furan has been well documented previously, and it was classified as "possible human carcinogen (Group 2B)" by the International Agency for Research on Cancer. Various pathways have been reported for the formation of furan, that is, thermal degradation and/or thermal rearrangement of carbohydrates in the presence of amino acids, thermal degradation of certain amino acids, including aspartic acid, threonine, ${\alpha}$-alanine, serine, and cysteine, oxidation of ascorbic acid at higher temperatures, and oxidation of polyunsaturated fatty acids and carotenoids. Owing to the complexity of the formation mechanism, a vast number of studies have been published on monitoring furan in commercial food products and on the potential strategies for reducing furan. Thus, we present a comprehensive review on the current status of commercial food monitoring databases and the possible furan reduction methods. Additionally, we review analytical methods for furan detection and the toxicity of furan.
In order to find out changes of chemical components related to browning of dried jujube, four varieties were subjected to the proximate analysis. Boeun, one of the major varieties in Korea, was sun-dried and stored for 12 months and analyzed periodically for one year. Browning, titrable acidity, pH, contents of ascorbic acid, organic acid, total amino acid, free amino acid, free sugar, hydroxymethylfurfural (HMF) and phenolic compounds were determined and compared with those of fresh jujube. The results obtained are summarized as follows; 1. In comparison with other similar fruits, jujube was high contents of ascorbic acid (62-79mg%) and carbohydrate (22-28%) excluding fiber. 2. Browning was increased in both steam-treated and nontreated plot together as storage period was prolonged. 3. Ascorbic acid content of fresh jujube was as high as 297.4mg% DB, but it was decreased to 20.2mg% DB, after 12 months storage. Therefore, loss of ascorbic acid was very great up to 93% of its original content. 4. Five kinds of organic acid, oxalic, succinic, fumaric, malic and citric acid were identified as major organic acids. It was interesting that only fumaric acid content was increased while others decreased during storage. 5. Seventeen kinds of amino acid were identified in the analysis of total amino acid content. Major amino acids were found to be proline, threonine, glutamic acid and lysine. During 12 months storage, 30% of original total amino acid was decreased and this was mostly accounted for free amino acids. 6. Threonine, proline, alanine and valine were identified as free amino acids which showed 85% decrease after 12 months storage. 7. Free sugars of jujube were composed of fructose, glucose and sucrose. They showed 24% decrease after 12 months storage, Especially sucrose was not detected after 4 months storage. 8. HMF of dried jujube was isolated and identified. Its content was increased up to great extent as storage period prolonged. 9. Caffeic, ferulic and P-coumaric acid were identified as jujube phenolic compounds which were significantly decreased during storage. In conclusion, non-enzymatic browning was thought to be more important than enzymatic reaction in the dried jujube. Amino-carbonyl reaction, ascorbic acid oxidation and reaction between phenolic compounds and sugar or nitrogen compounds were observed to be related to this browning.
Kim, Sun-Lim;Chi, Hee-Youn;Kim, Jung-Tae;Hur, On-Sook;Kim, Deog-Su;Suh, Sae-Jung;Kim, Hyun-Bok;Cheong, Ill-Min
KOREAN JOURNAL OF CROP SCIENCE
/
v.56
no.4
/
pp.349-360
/
2011
The objectives of present study were to characterize the peptides which were isolated from Korean fermented soybean paste, chungkukjang, and to determine their antioxidant activities. Four fractions were collected from the methanol extract of chungkukjang by using a recycling preparative HPLC. Among fractions, Fr-2 was identified to be highly potent free radical scavenging activity in the assay of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and nitroblue tetrazolium(NBT)-reduction inhibition. Base on antioxidant effects, fraction Fr-2 was employed for the refraction with a prep-column and separated into five fractions of which two fractions were identified to have higher antioxidant activity. To confirm the amino acid constituents of antioxidant fractions Fr-2-2 and Fr-2-3 were analyzed, and eight kinds of amino acids such as aspartic acid, threonine, serine, glutamic acid, glycine, lysine, histidine, and arginine were identified as the constituent amino acids. Antioxidant activities of the separated peptides were further assessed cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT), and fluorescence-activated cell sorting (FACS) analysis of H4IIE cells treated with hydrogen peroxide (H2O2). Chungkukjang peptides have shown their ability to protect H4IIE rat hepatoma cells against H2O2- induced oxidative stress by concentration and time-dependent manner. Therefore, These results indicated that fermented soybean paste chungkukjang will be promoted the antioxidant and radical scavenging activities, and beneficial for health. The antioxidant peptide fractions Fr-2-2 and Fr-2-3 were denominated as P-NICS-1 and P-NICS-2, respectively. However, further studies were required to clarify their amino acid sequences and molecular properties, and physiological significances.
Kim, Do Hyeon;Nguyen, Quyet Thang;Ko, Gyeong Soo;Yang, Jin Kuk
Journal of Microbiology and Biotechnology
/
v.30
no.12
/
pp.1905-1911
/
2020
Homoserine dehydrogenase (HSD) catalyzes the reversible conversion of ʟ-aspartate-4-semialdehyde to ʟ-homoserine in the aspartate pathway for the biosynthesis of lysine, methionine, threonine, and isoleucine. HSD has attracted great attention for medical and industrial purposes due to its recognized application in the development of pesticides and is being utilized in the large scale production of ʟ-lysine. In this study, HSD from Bacillus subtilis (BsHSD) was overexpressed in Escherichia coli and purified to homogeneity for biochemical characterization. We examined the enzymatic activity of BsHSD for ʟ-homoserine oxidation and found that BsHSD exclusively prefers NADP+ to NAD+ and that its activity was maximal at pH 9.0 and in the presence of 0.4 M NaCl. By kinetic analysis, Km values for ʟ-homoserine and NADP+ were found to be 35.08 ± 2.91 mM and 0.39 ± 0.05 mM, respectively, and the Vmax values were 2.72 ± 0.06 μmol/min-1 mg-1 and 2.79 ± 0.11 μmol/min-1 mg-1, respectively. The apparent molecular mass determined with size-exclusion chromatography indicated that BsHSD forms a tetramer, in contrast to the previously reported dimeric HSDs from other organisms. This novel oligomeric assembly can be attributed to the additional C-terminal ACT domain of BsHSD. Thermal denaturation monitoring by circular dichroism spectroscopy was used to determine its melting temperature, which was 54.8℃. The molecular and biochemical features of BsHSD revealed in this study may lay the foundation for future studies on amino acid metabolism and its application for industrial and medical purposes.
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