• Title/Summary/Keyword: Metabolic Engineering

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Analysis of soyasaponin content and biosynthesis-related gene expression in young pea (Pisum sativum L.) sprouts

  • Gang Deok Han;HanGyeol Lee;Jae-Hyeok Park;Young Jae Yun;Gee Woo Kim;Sangyun Jeong;So-Yeon Moon;Hye-Young Seo;Young-Cheon, Kim;Woo Duck Seo;Jeong Hwan Lee
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.70-75
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    • 2023
  • In legumes, soyasaponins, one of triterpenoid saponins, are major components of secondary metabolites with a more diverse array of bioactive chemicals. Although the biosynthetic pathway of soyasaponins has been largely studied in soybean, the study on the soyasaponin contents and biosynthesis-related gene expression in pea (Pisum sativum L.) is poorly understood. Here, we found the accumulation of only soyasaponin Bb component in the sprouts of two Korean domestic pea cultivars (Dachung and Sachul). This pattern was consistent with our observation that increased expression of PsUGT73P2 and PsUGT91H4 genes, but not PsCYP72A69, could be responsible for biosynthesis of only soyasaponin Bb in pea by examining their gene expression. However, gradual accumulation of soyasaponin Bb at developmental stages was not consistent with the expression of PsUGT73P2 and PsUGT91H4, suggesting that the changes of their protein activities may affect the accumulation patterns of soyasaponin Bb. We also revealed that the increased expression levels of PsUGT73P2 and PsUGT91H4 during light to dark transition led to increase of soyasaponin Bb contents. Collectively, our results provided a molecular basis of metabolic engineering for enhancing useful soyasaponin Bb metabolites in Korean domestic pea cultivars.

Constipation anti-aging effects by dairy-based lactic acid bacteria

  • Mohamad Hafis Jaafar;Pei Xu;Uma-Mageswary Mageswaran;Shandra-Devi Balasubramaniam;Maheswaran Solayappan;Jia-Jie Woon;Cindy Shuan-Ju Teh;Svetoslav Dimitrov Todorov;Yong-Ha Park;Guoxia Liu;Min-Tze Liong
    • Journal of Animal Science and Technology
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    • v.66 no.1
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    • pp.178-203
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    • 2024
  • Constipation, which refers to difficulties in defecation and infrequent bowel movement in emptying the gastrointestinal system that ultimately produces hardened fecal matters, is a health concern in livestock and aging animals. The present study aimed to evaluate the potential effects of dairy-isolated lactic acid bacteria (LAB) strains to alleviate constipation as an alternative therapeutic intervention for constipation treatment in the aging model. Rats were aged via daily subcutaneous injection of D-galactose (600 mg/body weight [kg]), prior to induction of constipation via oral administration of loperamide hydrochloride (5 mg/body weight [kg]). LAB strains (L. fermentum USM 4189 or L. plantarum USM 4187) were administered daily via oral gavage (1 × 10 Log CFU/day) while the control group received sterile saline. Aged rats as shown with shorter telomere lengths exhibited increased fecal bulk and soften fecal upon administration of LAB strains amid constipation as observed using the Bristol Stool Chart, accompanied by a higher fecal moisture content as compared to the control (p < 0.05). Fecal water-soluble metabolite profiles showed a reduced concentration of threonine upon administration of LAB strains compared to the control (p < 0.05). Histopathological analysis also showed that the administration of LAB strains contributed to a higher colonic goblet cell count as compared to the control (p < 0.05). The present study illustrates the potential of dairy-sourced LAB strains as probiotics to ameliorate the adverse effect of constipation amid aging, and as a potential dietary intervention strategy for dairy foods including yogurt and cheese.

A Study of Synthesis and Biological Function on DL-1-Aminoethylphosphonic Acid (DL-1-Aminoethylphosphonic acid의 생물학적(生物學的) 기능연구(機能硏究))

  • Kim, Sook-He;Cho, Jung-Nam;Kim, Yong-Joon
    • Journal of Nutrition and Health
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    • v.2 no.4
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    • pp.173-181
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    • 1969
  • Since 1959 ${\beta}-aminoethylphosphonic$ acid was discovered in the living organism, the biosynthesis and biological functions of aminophosphonic acids have been extensively studied. The author designed and carried out this study for 14 weeks to find out the metabolic function of Ethylaminophosphonic acid (AEP) and it's utilization in the living body. Sixty rats, thirty males and thirty females aged $40{\pm}5$ days were divided into two parts, one for alanine supplemented as control group and the other for AEP as experimental group to compare metabolic pathway of ordinary amino acid with that of AEP. Both alamine and AEP group were divived into two subgroups according to the level of supplements, 0.1% and 0.2% of the diet. The major components of the diet in this study were composed of 20% casein, 72% Sugar, 4% fat, 4% salt Mixture, and all kind of Uitamins in adeguate amount. For comparision of biological values between experimental and control group in terms of body weight, uninary nitrogen, creatinine excretion and final orgam weight, there were no statically significant difference in these respects. This meant AEP could be utilized in the body as much as alanine could. Urinary phosphorus excretion was determined by developing the blue color to read on the Spectronic 20. Statistically insignificance in the urinary phosphorus excretion between experimental and control group was observed in spite of the supplementation of phosphorus of AEP for experimental group in the diet. The level of blood phosphorus was higher in experimental group than that in control group this result supported above result. In the analysis of fat and nitrogen contents in the liver, AEP group showed slightly higher than control in both respects. But it was noteworthy 0.2% AEP group in both sex were higher than 0.1% AEP in liver fat content. Histological examinal of internal organs liiver, lung, spleen, heart, kindey, adrenal and sex organs showed no changes in all groups included in this study. The group supplemented higher level of diet. by alanine 0.2% and AEP 0.2% stayed on less body weight gain and lower liver weight. This result could be interpreted that amino acid imbalanced condition was arose in the body.

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Automatic Interpretation of Epileptogenic Zones in F-18-FDG Brain PET using Artificial Neural Network (인공신경회로망을 이용한 F-18-FDG 뇌 PET의 간질원인병소 자동해석)

  • 이재성;김석기;이명철;박광석;이동수
    • Journal of Biomedical Engineering Research
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    • v.19 no.5
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    • pp.455-468
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    • 1998
  • For the objective interpretation of cerebral metabolic patterns in epilepsy patients, we developed computer-aided classifier using artificial neural network. We studied interictal brain FDG PET scans of 257 epilepsy patients who were diagnosed as normal(n=64), L TLE (n=112), or R TLE (n=81) by visual interpretation. Automatically segmented volume of interest (VOI) was used to reliably extract the features representing patterns of cerebral metabolism. All images were spatially normalized to MNI standard PET template and smoothed with 16mm FWHM Gaussian kernel using SPM96. Mean count in cerebral region was normalized. The VOls for 34 cerebral regions were previously defined on the standard template and 17 different counts of mirrored regions to hemispheric midline were extracted from spatially normalized images. A three-layer feed-forward error back-propagation neural network classifier with 7 input nodes and 3 output nodes was used. The network was trained to interpret metabolic patterns and produce identical diagnoses with those of expert viewers. The performance of the neural network was optimized by testing with 5~40 nodes in hidden layer. Randomly selected 40 images from each group were used to train the network and the remainders were used to test the learned network. The optimized neural network gave a maximum agreement rate of 80.3% with expert viewers. It used 20 hidden nodes and was trained for 1508 epochs. Also, neural network gave agreement rates of 75~80% with 10 or 30 nodes in hidden layer. We conclude that artificial neural network performed as well as human experts and could be potentially useful as clinical decision support tool for the localization of epileptogenic zones.

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Metabolic impairment pattern analysis of the Alzheimer's disease (Alzheimer's Disease의 대사영상패턴 분석)

  • Juh, Ra-Hyeong;Lee, Chang-Uk;Chung, Yong-An;Choe, Bo-Young;Suh, Tae-Suk
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2004.11a
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    • pp.91-95
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    • 2004
  • Several MRI studies have reported reductions in temporal lobe volumes in Alzheimer's disease (AD). Measures have been usually obtained with regions-of-interest (ROI) drawn manually on selected medial and lateral portions of the temporal lobes, with variable choices of anatomical borders across different studies. We used the automated voxel-based morphometry (VBM) approach to investigate gray matter abnormalities over the entire extension of the temporal lobe in 10AD patients (MM5E 22)and 22 healthy controls. Foci of significantly reduced gray matter volume in AD patients were detected in both medial and lateral temporal regions, most significantly in the right and left posterior parahippocarmpal gyri. At a more flexible statistical threshold (P<0.01, uncorrected for multiple comparisons), circumscribed foci of significant gray matter reduction were also detected in the right amygdala/enthorinal cortex, the anterior and posterior borders of the superior temporal gyrus bilaterally, and the anterior portion of the left middle temporal gyrus. These VBM results confirm previous findings of temporal lobe atrophic changes in AD, and suggest that these abnormalities may be confined to specific sites within that lobe, rather than showing a widespread distribution.

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Estimation of Human Lower-Extremity Muscle Force Under Uncertainty While Rising from a Chair (의자에서 일어서는 동작 시 불확실성을 고려한 인체 하지부 근력 해석)

  • Jo, Young Nam;Kang, Moon Jeong;Chae, Je Wook;Yoo, Hong Hee
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.38 no.10
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    • pp.1147-1155
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    • 2014
  • Biomechanical models are often used to predict muscle and joint forces in the human body. For estimation of muscle forces, the body and muscle properties have to be known. However, these properties are difficult to measure and differ from person to person. Therefore, it is necessary to predict the change in muscle forces depending on the body and muscle properties. The objective of the present study is to develop a numerical procedure for estimating the muscle forces in the human lower extremity under uncertainty of body and muscle properties during rising motion from a seated position. The human lower extremity is idealized as a multibody system in which eight Hill-type muscle force models are employed. Each model has four degrees of freedom and is constrained in the sagittal plane. The eight muscle forces are determined by minimizing the metabolic energy consumption during the rising motion. Uncertainty analysis is performed using a first-order reliability method. The one-standard-deviation range of agonistic muscle forces is calculated to be about 150-300 N.

Biological Functions of the COOH-Terminal Amino Acids of the $\alpha$-Subunit of Tethered Equine Chorionic Gonadotropin

  • Jeoung, Youn-Hee;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.47-53
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    • 2010
  • Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

Effect of Methionine on Cephalosporin C Production in a Fluidized- bed Bioreactor (유동층 생물반응기에서 세파로스포린 C 생산에 대한 메치오닌의 영향)

  • Kim, Eui-Yong;Yoo, Young-Je;Park, Young-Hoon
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.611-618
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    • 1989
  • Effects of methionine on cephalosporin C(CPC) production in a fluidized-bed bioreactor were investigated using bioparticles of Cephalosporium acremonium. Since methionine was found to be an important metabolic regulator on the synthesis of cephalosporin C, the effects of its concentration in the cuture broth and feeding mode to the bioreactor were studied. It was observed that the presence of initial methionine was essential for higher cephalosporin C production and there existed an optimal content of methionine. Carbon consumption rate also increased significantly under the presence of methionine. Production of cephalosporin C was most active when methionine was exhausted in the broth; however its additional feeding did not enhance the antibiotic production in the fluidized-bed bioreactor as much as expected. It was therfore considered important to feed an optimal content of methionine at the early operating stage for a higher cephalosporin C production in a fluidized-bed bioreactor. An interesting thing to note was that titre of the antibiotic with reused bioparticles was about 2 times higher in the methionine containing medium than that without methionine. Therefore repeated use of bioparticles, with an optimal content of methionine, was believed to be very useful to enhance to process productivity.

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Production of Fructose 6-Phoschate from Starch Using Thermostable Enzymes (내열성 효소를 이용한 전분으로부터 6-인산과당의 제조)

  • Kwun, Kyu-Hyuk;Cha, Wol-Suk;Kim, Bok-Hee;Shin, Hyun-Jae
    • KSBB Journal
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    • v.22 no.5
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    • pp.345-350
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    • 2007
  • Phosphosugars are found in all living organisms and are commercially valuable compounds with possible applications in the development of a wide range of specialty chemicals and medicines. In carbohydrate metabolism, fructose 6-phosphate (F6P) is an essential intermediate formed by phosphorylation of 6' position of fructose in glycolysis, gluconeogenesis, pentose phosphate pathway and Calvin cycle. In glycolysis, F6P lies within the glycolysis metabolic pathway and is produced by isomerisation of glucose 6-phosphate. For large-scale production, F6P could be produced from starch using many enzymes such as pullulanase, starch phosphorylase, isomerase and mutase. In enzymatic reactions carried out at high temperatures, the solubility of starch is increased and microbial contamination is minimized. Thus, thermophile-derived enzymes are preferred over mesophile-derived enzymes for industrial applications using starch. Recently, we reported the production of glucose 1-phosphate (G1P) from starch by Thermus caldophilus GK24 enzymes. Here we report the production of F6P from starch through three steps; from starch to glucose 1-phosphate (glucan phosphorylase, GP), then glucose 6-phosphate (phosphoglucomutase, GM) and then F6P (phosphoglucoisomerase, GI). Using 200 L of 1.2% soluble starch solution in potassium phosphate buffer, 1,253 g of G1P were produced. Then, 30% yields of F6P were attained at the optimum reaction conditions of GM : G1 (1 : 2.3), 63.5$^{\circ}C$, and pH 6.85. The optimum conditions were found by response surface methodology and the theoretical values were confirmed by the experiments. The optimum starch concentrations were 20 g/L under the given conditions.

Thermophilic Biohydrogen Production from Glucose with a Long-term Operation of CSTR (CSTR의 장기운전을 통한 포도당으로부터의 고온 수소생산)

  • Ahn, Yeong-Hee;Oh, You-Kwan;Park, Sung-Hoon
    • KSBB Journal
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    • v.20 no.6
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    • pp.425-430
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    • 2005
  • Thermophilic $H_2$ was produced for 1 year using a bench-scale continuous stirred tank reactor(CSTR). The CSTR was inoculated with anaerobically digested sludge after heat treatment and fed with a glucose-based medium. The reactor showed relatively short start-up period(30 days) and high maximal $H_2$ yield(2.4 mol $H_2/mol$ glucose). Keeping pH 5.0 or less suppressed methanogenic activity. Bacteria affiliated with Thermoanaerobacterium thermosaccharolyticum kept being dominant from approximately 40 days as determined by DGGE. Environmental perturbation(pH or temperature) caused the decrease of biomass concentration in the reactor and the instability of reactor performance, $H_2$ production rate and $H_2$ yield. The unstable performance was accompanied with high concentration of lactate in the effluent. Taken together, the poor recovery of CSTR after perturbations could be partly explained by low biomass concentration and/or metabolic shift of the major population in the CSTR.