• Title/Summary/Keyword: enzyme selectivity

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Highly Efficient Biotransformation of Notoginsenoside R1 into Ginsenoside Rg1 by Dictyoglomus thermophilum β-xylosidase Xln-DT

  • Li, Qi;Wang, Lei;Fang, Xianying;Zhao, Linguo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.447-457
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    • 2022
  • Notoginsenoside R1 and ginsenoside Rg1 are the main active ingredients of Panax notoginseng, exhibiting anti-fatigue, anti-tumor, anti-inflammatory, and other activities. In a previous study, a GH39 β-xylosidase Xln-DT was responsible for the bioconversion of saponin, a natural active substance with a xylose group, with high selectivity for cleaving the outer xylose moiety of notoginsenoside R1 at the C-6 position, producing ginsenoside Rg1 with potent anti-fatigue activity. The optimal bioconversion temperature, pH, and enzyme dosage were obtained by optimizing the transformation conditions. Under optimal conditions (pH 6.0, 75℃, enzyme dosage 1.0 U/ml), 1.0 g/l of notoginsenoside R1 was converted into 0.86 g/l of ginsenoside Rg1 within 30 min, with a molar conversion rate of approximately 100%. Furthermore, the in vivo anti-fatigue activity of notoginsenoside R1 and ginsenoside Rg1 were compared using a suitable rat model. Compared with the control group, the forced swimming time to exhaustion was prolonged in mice by 17.3% in the Rg1 high group (20 mg/kg·d). Additionally, the levels of hepatic glycogen (69.9-83.3% increase) and muscle glycogen (36.9-93.6% increase) were increased. In the Rg1 group, hemoglobin levels were also distinctly increased by treatment concentrations. Our findings indicate that treatment with ginsenoside Rg1 enhances the anti-fatigue effects. In this study, we reveal a GH39 β-xylosidase displaying excellent hydrolytic activity to produce ginsenoside Rg1 in the pharmaceutical and food industries.

A prognosis discovering lethal-related genes in plants for target identification and inhibitor design (식물 치사관련 유전자를 이용하는 신규 제초제 작용점 탐색 및 조절물질 개발동향)

  • Hwang, I.T.;Lee, D.H.;Choi, J.S.;Kim, T.J.;Kim, B.T.;Park, Y.S.;Cho, K.Y.
    • The Korean Journal of Pesticide Science
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    • v.5 no.3
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    • pp.1-11
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    • 2001
  • New technologies will have a large impact on the discovery of new herbicide site of action. Genomics, combinatorial chemistry, and bioinformatics help take advantage of serendipity through tile sequencing of huge numbers of genes or the synthesis of large numbers of chemical compounds. There are approximately $10^{30}\;to\;10^{50}$ possible molecules in molecular space of which only a fraction have been synthesized. Combining this potential with having access to 50,000 plant genes in the future elevates tile probability of discovering flew herbicidal site of actions. If 0.1, 1.0 or 10% of total genes in a typical plant are valid for herbicide target, a plant with 50,000 genes would provide about 50, 500, and 5,000 targets, respectively. However, only 11 herbicide targets have been identified and commercialized. The successful design of novel herbicides depends on careful consideration of a number of factors including target enzyme selections and validations, inhibitor designs, and the metabolic fates. Biochemical information can be used to identify enzymes which produce lethal phenotypes. The identification of a lethal target site is an important step to this approach. An examination of the characteristics of known targets provides of crucial insight as to the definition of a lethal target. Recently, antisense RNA suppression of an enzyme translation has been used to determine the genes required for toxicity and offers a strategy for identifying lethal target sites. After the identification of a lethal target, detailed knowledge such as the enzyme kinetics and the protein structure may be used to design potent inhibitors. Various types of inhibitors may be designed for a given enzyme. Strategies for the selection of new enzyme targets giving the desired physiological response upon partial inhibition include identification of chemical leads, lethal mutants and the use of antisense technology. Enzyme inhibitors having agrochemical utility can be categorized into six major groups: ground-state analogues, group specific reagents, affinity labels, suicide substrates, reaction intermediate analogues, and extraneous site inhibitors. In this review, examples of each category, and their advantages and disadvantages, will be discussed. The target identification and construction of a potent inhibitor, in itself, may not lead to develop an effective herbicide. The desired in vivo activity, uptake and translocation, and metabolism of the inhibitor should be studied in detail to assess the full potential of the target. Strategies for delivery of the compound to the target enzyme and avoidance of premature detoxification may include a proherbicidal approach, especially when inhibitors are highly charged or when selective detoxification or activation can be exploited. Utilization of differences in detoxification or activation between weeds and crops may lead to enhance selectivity. Without a full appreciation of each of these facets of herbicide design, the chances for success with the target or enzyme-driven approach are reduced.

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Evaluation of a Chromogenic Medium Supplemented with Glucose for Detecting Enterobacter sakazakii

  • Song, Kwang-Young;Hyeon, Ji-Yeon;Shin, Ho-Chul;Park, Chan-Kyu;Choi, In-Soo;Seo, Kun-Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.579-584
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    • 2008
  • A commercial chromogenic agar medium (DFI) was supplemented with glucose (mDFI) to enhance the specificity of Enterobacter sakazakii (E. sakazakit) detection. Escherichia vulneris (E. vulneris), a putative false-positive strain on the DFI medium, produces ${\alpha}$-glucosidase. The enzyme ${\alpha}$-glucosidase hydrolyzes a substrate, 5-bromo-4-chloro-3-indolyl-${\alpha}$, D-glucopyranoside $(X{\alpha}Glc)$, producing green colonies. E. sakazakii strains produced green colonies on both DFI and mDFI agar, whereas E. vulneris produced green colonies on DFI agar but small white colonies on mDFI agar. E. sakazakii and E. vulneris were also readily differentiated by colony color when the mixed culture of the two strains was plated on mDFI agar and incubated for 24 h at $37^{\circ}C$. The results indicate that the selectivity of the commercial chromogenic agar medium could be improved by a simple supplementation with glucose.

Improving Catalytic Efficiency and Changing Substrate Spectrum for Asymmetric Biocatalytic Reductive Amination

  • Jiang, Wei;Wang, Yali
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.146-154
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    • 2020
  • With the advantages of biocatalytic method, enzymes have been excavated for the synthesis of chiral amino acids by the reductive amination of ketones, offering a promising way of producing pharmaceutical intermediates. In this work, a robust phenylalanine dehydrogenase (PheDH) with wide substrate spectrum and high catalytic efficiency was constructed through rational design and active-site-targeted, site-specific mutagenesis by using the parent enzyme from Bacillus halodurans. Active sites with bonding substrate and amino acid residues surrounding the substrate binding pocket, 49L-50G-51G, 74M,77K, 122G-123T-124D-125M, 275N, 305L and 308V of the PheDH, were identified. Noticeably, the new mutant PheDH (E113D-N276L) showed approximately 6.06-fold increment of kcat/Km in the oxidative deamination and more than 1.58-fold in the reductive amination compared to that of the wide type. Meanwhile, the PheDHs exhibit high capacity of accepting benzylic and aliphatic ketone substrates. The broad specificity, high catalytic efficiency and selectivity, along with excellent thermal stability, render these broad-spectrum enzymes ideal targets for further development with potential diagnostic reagent and pharmaceutical compounds applications.

Garcinexanthone G, a Selective Butyrylcholinesterase Inhibitor from the Stem Bark of Garcinia atroviridis

  • Khaw, Kooi-Yeong;Murugaiyah, Vikneswaran;Khairuddean, Melati;Tan, Wen-Nee
    • Natural Product Sciences
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    • v.24 no.2
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    • pp.88-92
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    • 2018
  • The present study was undertaken to investigate the isolated compounds from the stem bark of Garcinia atroviridis as potential cholinesterase inhibitors and the ligand-enzyme interactions of selected bioactive compounds in silico. The in vitro cholinesterase results showed that quercetin (3) was the most active AChE inhibitor ($12.65{\pm}1.57{\mu}g/ml$) while garcinexanthone G (6) was the most active BChE inhibitor ($18.86{\pm}2.41{\mu}g/ml$). It is noteworthy to note that compound 6 was a selective inhibitor with the selectivity index of 11.82. Molecular insight from docking interaction further substantiate that orientation of compound 6 in the catalytic site which enhanced its binding affinity as compared to other xanthones. The nature of protein-ligand interactions of compound 6 is mainly hydrogen bonding, and the hydroxyl group of compound 6 at C-10 is vital in BChE inhibition activity. Therefore, compound 6 is a notable lead for further drug design and development of BChE selective inhibitor.

Glucose Sensing Properties of Electrospinning-Synthesized ZnO Nanofibers (전기방사로 합성된 산화아연 나노섬유의 Glucose 감응특성)

  • Choi, Jong-Myoung;Byun, Joon-Hyuk;Kim, Sang Sub
    • Korean Journal of Materials Research
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    • v.25 no.12
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    • pp.655-658
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    • 2015
  • The development of glucose biosensors has been attracting much attention because of their importance in monitoring glucose in the human body; such sensors are used to diagnose diabetes and related human diseases. Thanks to the high selectivity, sensitivity to glucose detection, and relatively low-cost fabrication of enzyme-immobilized electrochemical glucose sensors, these devices are recognized as one of the most intensively investigated glucose sensor types. In this work, ZnO nanofibers were synthesized using an electrospinning method with polyvinyl alcohol zinc acetate as precursor material. Using the synthesized ZnO nanofibers, we fabricated glucose biosensors in which glucose oxidase was immobilized on the ZnO nanofibers. The sensors were used to detect a wide range of glucose from 10 to 700 M with a sensitivity of $10.01nA/cm^2-{\mu}M$, indicating that the ZnO nanofiber-based glucose sensor can be used for the detection of glucose in the human body. The control of nanograins in terms of the size and crystalline quality of the individual nanofibers is required for improving the glucose-sensing abilities of the nanofibers.

Electrochemical Conversion of Carbon Dioxide (이산화탄소의 전기화학적 변환)

  • Song, Ji-Eun;Shin, Woon-Sup
    • Journal of the Korean Electrochemical Society
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    • v.12 no.2
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    • pp.131-141
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    • 2009
  • The conversion of carbon dioxide to value-added compounds has been attracted to solve the environmental problems due to the climate change caused by greenhouse effect in addition to recycle the abundant and renewable carbon source. For utilizing carbon dioxide to useful compounds, the development of catalysts and optimization of experimental conditions are indispensable since carbon dioxide is the most stable one among carbon compounds and the a certain amount of energy is required for the carbon dioxide conversion. The technologies developed for the electrochemical carbon dioxide conversion were reviewed in terms of electrocatalyst which can be electrode material, inorganic complex, and enzyme. This field should be developed further since no good catalyst having selectivity, efficiency, and stability all together.

Organic-Inorganic Hybrid Nanoflowers as Potent Materials for Biosensing and Biocatalytic Applications

  • Tran, Tai Duc;Kim, Moon Il
    • BioChip Journal
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    • v.12 no.4
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    • pp.268-279
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    • 2018
  • Flower-shaped organic-inorganic hybrid nanostructures, termed nanoflowers, have received considerable recent attention as they possess greatly enhanced activity, stability, durability, and even selectivity of entrapped organic biomolecules, which are much better than those from the conventional methods. They can be synthesized simply via co-incubation of organic and inorganic components in aqueous buffer at room temperature and yield hierarchical nanostructures with large surface-to-volume ratios, allowing for low-cost production by easy scale-up, as well as the high loading capacity of biomolecules without severe mass transfer limitations. Since a pioneering study reported on hybrid nanoflowers prepared with protein and copper sulfate, many other organic and inorganic components, which endow nanoflowers with diverse functionalities, have been employed. Thanks to these features, they have been applied in a diverse range of areas, including biosensors and biocatalysis. To highlight the progress of research on organic-inorganic hybrid nanoflowers, this review discusses their synthetic methods and mechanisms, structural and biological characteristics, as well as recent representative applications. Current challenges and future directions toward the design and development of multi-functional nanoflowers for their widespread utilization in biotechnology are also discussed.

Biosynthesis of Compound K, a biologically active saponin of ginseng(Panax ginseng) by bioconversion (인삼(Panax ginseng)으로부터 생물전환을 이용한 생리활성물질인 Compound K의 생합성)

  • Kim, MooSung;Kim, Ja-i;Jung, Kyung-Hwan;Yu, Kwang-Won;Moon, Gi-Seong;Lee, Hyang-Yeol
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.5
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    • pp.1335-1344
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    • 2021
  • Ginsenoside Compound K is a triterpene saponin found in the leafs, stems and roots of Panax ginseng. This study aimed to prepare a valuable ginsenoside Compound K using ginseng extracts with the enzyme(Plantase). Plantase showed very efficient activity to produce Compound K from ginseng extracts. Plantase exhibited the highest activity at pH 5 and 50 ℃, as a result of investigating the yield of Compound K by changing the temperature and pH, while fixing the enzyme concentration to 10% or 15% over 48 hours of reaction time. Under optimium conditions, Plantase produced and accumulated Compound K over 35 wt% of whole ginseng extracts. Antimicrobial activitiy of bioconvertied ginseng extracts showed selectivity against Cutibacterium acnes KCTC 3314. Minimal inhibitory concentration (MIC) of bioconverted ginseng extract (35% of Compound K enriched extract) against Cutibacterium acnes KCTC 3314 strain is 31.25ug/mL. These results suggest that the Compound K enriched extract is potential materials for cosmetic products and Plantase is a very useful enzyme for Compound K production.

Selectivity of Bleaching Herbicides Caused by Physiological Differences between Rice and Barnyardgrass (벼와 피의 생리적(生理的) 차이(差異)에 의한 백화형(白化型) 제초제(除草劑)의 선택성(選擇性))

  • Na, J.Y.;Kim, J.S.;Kim, T.J.;Cho, K.Y.;Pyon, J.Y.
    • Korean Journal of Weed Science
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    • v.12 no.2
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    • pp.89-101
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    • 1992
  • In this study, various physiological and biochemical experiments were conducted to know whether the selectivity between rice and barnyardgrass treated with bleaching herbicides containing diphenyl ether compounds was also partly based on their basic physiological proterties such as peroxidation ability, membrane stability or antioxidant system. It seemed to be partly based on the differences of their physiological characteristics that barnyardgrass was commonly more susceptible to most of the bleaching herbicides than rice. The scenescence of intact leaf segment was more rapid in barnyardgrass than in rice, indicating that barnyardgrass is weak at early stage. Also pigment metabolic ability, antioxidant enzyme activities(peroxidase, catalase, superoxide dismutase, glutathione reductase) and antioxidant content (tocopherol, ascorbic acid, glutathione, carotenoids) were lower in barnyardgrass on the basic of fresh weight. However, lipoxygenase activity and the content of unsaturated fatty acid which is vulnerable to oxygen radicals were higher in barnyardgrass, suggesting that barnyardgrass seedling bave a properties easy to be peroxidized. The differences of PPIX (protoporphyrin IX) or carotenoid content, which are the primary substances inducing herbicide activity, were not related to the selectivity between rice and barnyardgrass.

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