• 제목/요약/키워드: biochemical conversion

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Characterization of Gel16 as a Cytochrome P450 in Geldanamycin Biosynthesis and in-silico Analysis for an Endogenous Electron Transport System

  • Rimal, Hemraj;Yu, Sang-Cheol;Lee, Byeongsan;Hong, Young-Soo;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • 제29권1호
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    • pp.44-54
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    • 2019
  • Geldanamycin and its derivatives, inhibitors of heat shock protein 90, are considered potent anticancer drugs, although their biosynthetic pathways have not yet been fully elucidated. The key step of conversion of 4,5-dihydrogeldanamycin to geldanamycin was expected to catalyze by a P450 monooxygenase, Gel16. The adequate bioconversions by cytochrome P450 mostly rely upon its interaction with redox partners. Several ferredoxin and ferredoxin reductases are available in the genome of certain organisms, but only a few suitable partners can operate in full efficiency. In this study, we have expressed cytochrome P450 gel16 in Escherichia coli and performed an in vitro assay using 4,5-dihydrogeldanamycin as a substrate. We demonstrated that the in silico method can be applicable for the efficient mining of convenient endogenous redox partners (9 ferredoxins and 6 ferredoxin reductases) against CYP Gel16 from Streptomyces hygroscopicus. The distances for ligand FDX4-FDR6 were found to be $9.384{\AA}$. Similarly, the binding energy between Gel16-FDX4 and FDX4-FDR6 were -611.88 kcal/mol and -834.48 kcal/mol, respectively, suggesting the lowest distance and binding energy rather than other redox partners. These findings suggest that the best redox partners of Gel16 could be NADPH ${\rightarrow}$ FDR6 ${\rightarrow}$ FDX4 ${\rightarrow}$ Gel16.

Enzymatic Characterization and Comparison of Two Steroid Hydroxylases CYP154C3-1 and CYP154C3-2 from Streptomyces Species

  • Subedi, Pradeep;Kim, Ki-Hwa;Hong, Young-Soo;Lee, Joo-Ho;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • 제31권3호
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    • pp.464-474
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    • 2021
  • Bacterial cytochrome P450 (CYP) enzymes are responsible for the hydroxylation of diverse endogenous substances with a heme molecule used as a cofactor. This study characterized two CYP154C3 proteins from Streptomyces sp. W2061 (CYP154C3-1) and Streptomyces sp. KCCM40643 (CYP154C3-2). The enzymatic activity assays of both CYPs conducted using heterologous redox partners' putidaredoxin and putidaredoxin reductase showed substrate flexibility with different steroids and exhibited interesting product formation patterns. The enzymatic characterization revealed good activity over a pH range of 7.0 to 7.8 and the optimal temperature range for activity was 30 to 37℃. The major product was the C16-hydroxylated product and the kinetic profiles and patterns of the generated hydroxylated products differed between the two enzymes. Both enzymes showed a higher affinity toward progesterone, with CYP154C3-1 demonstrating slightly higher activity than CYP154C3-2 for most of the substrates. Oxidizing agents (diacetoxyiodo) benzene (PIDA) and hydrogen peroxide (H2O2) were also utilized to actively support the redox reactions, with optimum conversion achieved at concentrations of 3 mM and 65 mM, respectively. The oxidizing agents affected the product distribution, influencing the type and selectivity of the CYP-catalyzed reaction. Additionally, CYP154C3s also catalyzed the C-C bond cleavage of steroids. Therefore, CYP154C3s may be a good candidate for the production of modified steroids for various biological uses.

뇌척수액과 말초혈액 내 알츠하이머병의 생화학적 생체표지자 (Biochemical Biomarkers for Alzheimer's Disease in Cerebrospinal Fluid and Peripheral Blood)

  • 이영민;최원정;박민선;김어수
    • 노인정신의학
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    • 제16권1호
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    • pp.17-23
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    • 2012
  • The diagnosis of Alzheimer's disease (AD) is still obscure even to specialists. To improve the diagnostic accuracy, to find at-risk people as early as possible, to predict the efficacy or adverse reactions of pharmacotherapy on an individual basis, to attain more reliable results of clinical trials by recruiting better defined participants, to prove the disease-modifying ability of new candidate drugs, to establish prognosis-based therapeutic plans, and to do more, is now increasing the need for biomarkers for AD. Among AD-related biochemical markers, cerebrospinal beta-amyloid and tau have been paid the most attention since they are materials directly interfacing the brain interstitium and can be obtained through the lumbar puncture. Level of beta-amyloid is reduced whereas tau is increased in cerebrospinal fluid of AD patients relative to cognitively normal elderly people. Remarkably, such information has been found to help predict AD conversion of mild cognitive impairment. Despite inconsistent findings from previous studies, plasma beta-amyloid is thought to be increased before the disease onset, but show decreasing change as the disease progress. Regarding other peripheral biochemical markers, omics tools are being widely used not only to find useful biomarkers but also to generate novel hypotheses for AD pathogenesis and to lead new personalized future medicine.

Ionic Liquid Pretreatment of Lignocellulosic Biomass

  • Han, Song-Yi;Park, Chan-Woo;Kwon, Gu-Joong;Kim, Nam-Hun;Kim, Jin-Chul;Lee, Seung-Hwan
    • Journal of Forest and Environmental Science
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    • 제36권2호
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    • pp.69-77
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    • 2020
  • Lignocellulosic biomass has recalcitrant characteristics against chemical and biological conversion due to its structural heterogeneity and complexity. The pretreatment process to overcome these recalcitrant properties is essential, especially for the biochemical conversion of lignocellulosic biomass. In recent years, pretreatment methods using ionic liquids (ILs) and deep eutectic solvents (DESs) as the green solvent has attracted great attention because of their advantages such as easy recovery, chemical stability, temperature stability, nonflammability, low vapor pressure, and wide liquids range. However, there are some limitations such as high viscosity, poor economical feasibility, etc. to be solved for practical use. This paper reviewed the research activities on the pretreatment effect of various ILs including DESs and their co-solvents with organic solvents on the enzymatic saccharification efficiency of lignocellulosic biomass and the nanocellulose preparation from the pretreated products.

Acinetobacter sp. A54에 의한 Arabian Light 원유의 분해

  • 이창호;김희식;서현효;최성훈;오희목;윤병대
    • 한국미생물·생명공학회지
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    • 제25권5호
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    • pp.520-526
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    • 1997
  • Bacterial strains which degrade Arabian Light crude oil were isolated by enrichment culture from oil-spilled soil. The strain A54 was finally selected after testing emulsifying activity and oil conversion rate. Strain A54 was identified as a Acinetobacter sp. based on the morphological, biochemical and physiological characteristics. It appears to be highly specialized for growth on Arabian Light crude oil in minimal salts medium since it showed preference for oil or degradation products as substrates for growth. It was found that it could grow on at least fifteen different hydrocarbons. The optimum cultural and environmental conditions were as follows; 25$\circ$C for temperature, 7,5 for pH, 2.0% for NaCl concentration and 2.0% for crude oil concentration. Additionally, the optimal concentration of NH$_{4}$NO$_{3}$, and K$_{2}$HPO$_{4}$, were 12.5 mM and 0.057 mM, respectively. Cell growth and emulsifying activity as a function of time were also determined. Crude oil degradation and the reduction of product peaks were identified by the analysis of remnant oil by gas chromatography. Approximately 63% of crude oil were converted into a form no longer extractable by mixed organic solvents.

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Prions and Prion Diseases: Fundamentals and Mechanistic Details

  • Ryou, Chong-Suk
    • Journal of Microbiology and Biotechnology
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    • 제17권7호
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    • pp.1059-1070
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    • 2007
  • Prion diseases, often called transmissible spongiform encephalopathies (TSEs), are infectious diseases that accompany neurological dysfunctions in many mammalian hosts. Prion diseases include Creutzfeldt-Jakob disease (CJD) in humans, bovine spongiform encephalopathy (BSE, "mad cow disease") in cattle, scrapie in sheep, and chronic wasting disease (CWD) in deer and elks. The cause of these fatal diseases is a proteinaceous pathogen termed prion that lacks functional nucleic acids. As demonstrated in the BSE outbreak and its transmission to humans, the onset of disease is not limited to a certain species but can be transmissible from one host species to another. Such a striking nature of prions has generated huge concerns in public health and attracted serious attention in the scientific communities. To date, the potential transmission of prions to humans via foodborne infection and iatrogenic routes has not been alleviated. Rather, the possible transmission of human to human or cervids to human aggravates the terrifying situation across the globe. In this review, basic features about prion diseases including clinical and pathological characteristics, etiology, and transmission of diseases are described. Based on recently accumulated evidences, the molecular and biochemical aspects of prions, with an emphasis on the molecular interactions involved in prion conversion that is critical during prion replication and pathogenesis, are also addressed.

바이오에너지로의 전환을 위한 캘리포니아 식품가공공장 오.폐수 특성 조사 및 경제성 분석 (Survey and Economic Analysis of Food Industry Residues for Biomass-to-energy Conversion in Merced and Stanislaus Counties, California, USA)

  • 김대현
    • Journal of Biosystems Engineering
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    • 제34권4호
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    • pp.243-253
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    • 2009
  • This research expands investigations into the biomass resource potential associated with California's food processing industry by surveying industries within a two county region in the San Joaquin Valley, California, USA. A previous survey conducted in 2005 for the Sacramento Municipal Utility District (SMUD) quantified residue and waste generation from food processors and food preparation businesses in the Sacramento region. The present survey investigates residue and waste streams from food processors located in Merced and Stanislaus Counties. Sixty food processors were identified to participate in the survey, of which 49 responded (82%) and data were acquired for 38 (63%) (6 facilities closed or moved, 8 decided not to participate). Within the two counties, total annual waste among survey respondents amounted to 24,044 dry tons of high moisture (${\geq}$60%) food residuals, 5,358 dry tons of low moisture (<60%) food residuals; and 23.7 million $m^3$ of wastewater containing 38,814 tons of biochemical oxygen demand ($BOD_5$). The total potential electric power generation from these food residues was estimated at approximately $7\;MW_e$. Total solid waste resource included in the survey response was estimated at about 10% of statewide residue generation for processors falling within the Standard Industrial Classification (SIC) System Major Group 20 (Food and Kindred Products) categories.

Effect of Supplementing Betaine on Performance, Carcass Traits and Immune Responses in Broiler Chicken Fed Diets Containing Different Concentrations of Methionine

  • Rao, S.V. Rama;Raju, M.V.L.N.;Panda, A.K.;Saharia, Poonam;Sunder, G. Shyam
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권5호
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    • pp.662-669
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    • 2011
  • An experiment was conducted with broiler (Cobb 400) male chicks (n = 480) to determine the effect of betaine (Bet) supplementation (0 and 800 mg/kg) to diets containing five concentrations (15, 18, 20, 22 and 24 g/kg crude protein, CP) of methionine (Met) in a $2{\times}5$ factorial study for performance, carcass traits, immune responses, and serum parameters. Each diet was fed ad libitum from 1 to 42 d of age to 8 replicates of 6 chicks. Birds were housed in battery brooders placed in an open-sided poultry shed. Body weight gain, feed intake, feed conversion efficiency and slaughter variables were recorded at 21 and 42 d of age. Serum biochemical profile, antibody production against Newcastle disease (ND) and lymphocyte proliferation ratio (LPR) were analysed at 42 d of age. Supplementing Bet to diets containing sub-optimal concentrations of Met (15 g/kg) improved weight gain and breast yield at 21 d of age (p<0.01), and feed conversion efficiency at 42 d of age (p<0.05). Feed efficiency at 21 d of age, body weight gain at 42 d of age, slaughter variables except breast yield at 21 d of age and ND antibody titres were not affected (p>0.05) by the interaction. LPR increased (p<0.05) with Bet supplementation at 20 g Met/kg CP equal to those broilers fed 24 g Met without Bet. Bet supplementation enhanced the concentrations of protein, globulin and cholesterol in serum of broilers fed sub-optimal concentrations of Met. Results suggested that Bet supplementation (800 mg/kg diet) enhanced growth (21 d), feed conversion efficiency (42 d), breast yield and lymphocyte proliferation in broilers fed a diet containing 15 g Met/kg CP.

The Bioenergy Conversion Characteristics of Feedlot Manure Discharging from Beef Cattle Barn

  • Oh, Seung-Yong;Kim, Chang-Hyun;Yoon, Young-Man
    • 한국토양비료학회지
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    • 제48권6호
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    • pp.697-704
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    • 2015
  • This study was carried out to assess bioenergy conversion efficiency by biogas and solid fuel production in the cattle feedlot manure discharged from beef cattle barn. Feedlot manure was sampled from the cattle farmhouse located in Yong-in, Gyeonggi during the mid-fattening stage, periodically. The chemical characteristics, BMP (Biochemical methane potential) and HV (Heating values) of feedlot cattle manures were analyzed. Total solid contents of cattle feedlot manure were in the range of 29.98~44.28%, and volatile solid contents were in the range of 23.53~24.47%. In the anaerobic digestion of cattle feedlot manure, the methane production potential has increased from 0.141 to $0.187Nm^3kg^{-1}-VS_{added}$. The methane production of fresh cattle feedlot manure showed the range $0.141{\sim}0.187Nm^3kg^{-1}$-Manure (average $0.047Nm^3kg^{-1}$-Manure), the LHVs (lower heating values) of the produced methane were in the range of $316{\sim}560kcalkg^{-1}$-Manure (average $400kcalkg^{-1}$-Manure). In the direct combustion of fresh cattle feedlot manure, the LHVs were measured in the range of $747{\sim}1,271kcalkg^{-1}$-Manure (average $916kcalkg^{-1}$-Manure), and LHVs of solid fuel which have the water content of 20% were in the range of $2,694{\sim}2,876kcalkg^{-1}$-Manure (average $2,791kcalkg^{-1}$-Manure). Then, the drying energy of average $443kcalkg^{-1}$-Manure was consumed in the production of solid fuel which has a water content of 20%. Therefore, the direct combustion of cattle feedlot manure showed about 2.3 times higher LHV than the LHV of methane produced by anaerobic digestion. And LHV of solid fuel was about 6.0 times higher than the LHV of methane produced by anaerobic digestion. Then, the production of solid fuel presented more bioenergy conversion efficiency than the biogas production in the bioenergy use of cattle feedlot manure.

STRUCTURAL ANALYSIS OF THE SOLUBLE FORM OF BOVINE DOPAMINE BETA-HYDROXYLASE

  • Hwang, On-You;Joh, Tong-Hyub
    • Toxicological Research
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    • 제6권1호
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    • pp.99-107
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    • 1990
  • Dopamine beta-hydroxylase (DBH) is a neurotransmitter biosynthetic enzyme which catalyzes the conversion of dopamine to norepinephrine. Although structural studies of the mature form of this enzyme have been extensive, the culmination of these finding had been hightly controversial and contradictory. In this study, biochemical approaches were taken to characterize the structure of mature DBH. Soluble bovine DBH was purified from aderenal medulla. Three bands of 69 kDa, 72 kDa and 75 kDa which were physically separable and similar in structure were observed by SDS-PAGE. Furthermore, gas phase sequence analysis revealed that the 72 kDa band consists of two polypeptides which are present at equimolar concentrations and differed in that one had three extra amino acids at the N-terminus. Taken together, the soluble form of DBH exist in at least four forms, three identified by SDS-PAGE, one of which consists of two polypeptides as identified by N-terminal sequence analysis. The significance of these forms and their possible biosynthetic mechanisms are discussed.

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