• Title/Summary/Keyword: Module sequence

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Genenation of structural diversity in polyketides by combinatorial biosynthesis of polyketides: Part I. Generation of multiple bioactive macrolides by hybrid modular polyketide synthases in Streptomyces venezuelae, Part II. Production of novel rifamycins by combinatorial biosynthesis

  • Yoon, Yeo-Joon
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2002.10a
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    • pp.18-25
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    • 2002
  • The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

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THE FINITE DIMENSIONAL PRIME RINGS

  • Koh, Kwangil
    • Bulletin of the Korean Mathematical Society
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    • v.20 no.1
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    • pp.45-49
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    • 1983
  • If R is ring and M is a right (or left) R-module, then M is called a faithful R-module if, for some a in R, x.a=0 for all x.mem.M then a=0. In [4], R.E. Johnson defines that M is a prime module if every non-zero submodule of M is faithful. Let us define that M is of prime type provided that M is faithful if and only if every non-zero submodule is faithful. We call a right (left) ideal I of R is of prime type if R/I is of prime type as a R-module. This is equivalent to the condition that if xRy.subeq.I then either x.mem.I ro y.mem.I (see [5:3:1]). It is easy to see that in case R is a commutative ring then a right or left ideal of a prime type is just a prime ideal. We have defined in [5], that a chain of right ideals of prime type in a ring R is a finite strictly increasing sequence I$_{0}$.contnd.I$_{1}$.contnd....contnd.I$_{n}$; the length of the chain is n. By the right dimension of a ring R, which is denoted by dim, R, we mean the supremum of the length of all chains of right ideals of prime type in R. It is an integer .geq.0 or .inf.. The left dimension of R, which is denoted by dim$_{l}$ R is similarly defined. It was shown in [5], that dim$_{r}$R=0 if and only if dim$_{l}$ R=0 if and only if R modulo the prime radical is a strongly regular ring. By "a strongly regular ring", we mean that for every a in R there is x in R such that axa=a=a$^{2}$x. It was also shown that R is a simple ring if and only if every right ideal is of prime type if and only if every left ideal is of prime type. In case, R is a (right or left) primitive ring then dim$_{r}$R=n if and only if dim$_{l}$ R=n if and only if R.iden.D$_{n+1}$ , n+1 by n+1 matrix ring on a division ring D. in this paper, we establish the following results: (1) If R is prime ring and dim$_{r}$R=n then either R is a righe Ore domain such that every non-zero right ideal of a prime type contains a non-zero minimal prime ideal or the classical ring of ritght quotients is isomorphic to m*m matrix ring over a division ring where m.leq.n+1. (b) If R is prime ring and dim$_{r}$R=n then dim$_{l}$ R=n if dim$_{l}$ R=n if dim$_{l}$ R<.inf. (c) Let R be a principal right and left ideal domain. If dim$_{r}$R=1 then R is an unique factorization domain.TEX>R=1 then R is an unique factorization domain.

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Development of a New Automatic Image Quality Optimization System for Mobile TFT-LCD Applications (모바일 TFT-LCD 응용을 위한 새로운 형태의 자동화질 최적화 시스템 개발)

  • Ryu, Jee-Youl;Noh, Seok-Ho
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.47 no.1
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    • pp.17-28
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    • 2010
  • This paper presents a new automatic TFT-LCD image quality optimization system using DSP for the first time. Since conventional manual method depends on experiences of LCD module developers, it is highly labor-intensive and requires several correction steps providing large gamma correction error. The proposed system optimizes automatically gamma adjustment and power setting registers in mobile TFT-LCD driver IC to reduce gamma correction error, adjusting time, and flicker. It contains module-under-test (MUT, TFT-LCD module), PC installed with program, multimedia display tester for measuring luminance and flicker, and control board for interface between PC and TFT-LCD module. We have developed a new algorithm using 6-point programmable matching technique with reference gamma curve and applying automatic power setting sequence. Developed algorithm and program are generally applicable for most of the TFT-LCD modules. It is realized to calibrate gamma values of 1.8, 2.0, 2.2 and 3.0, and reduce flicker level. The control board is designed with DSP and FPGA, and it supports various interfaces such as RGB and CPU. Developed automatic image quality optimization system showed significantly reduced gamma adjusting time, reduced flicker, and much less average gamma error than conventional manual method. We believe that the proposed system is very useful to provide high-quality TFT-LCD and to improve developing process using optimized gamma-curve setting and automatic power setting.

Pickprimer: A Graphic User Interface Program for Primer Design on the Gene Target Region (픽프라이머 : 유전자 목표 구간 탐색 모듈을 포함한 프라이머 제작 그래픽 프로그램)

  • Chung, Hee;Mun, Jeong-Hwan;Lee, Seung-Chan;Yu, Hee-Ju
    • Horticultural Science & Technology
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    • v.29 no.5
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    • pp.461-466
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    • 2011
  • In genetic and molecular breeding studies of plants, researchers need to design various kinds of primers based on their research purposes. So far many kinds of web- or script-based non-commercial programs for primer design are available. Because most of them do not include user interface for multipurpose usage including gene structure prediction and direct target selection on sequences, it has been a laborious work to design primers targeting on the exon or intron regions of interesting genes. Here we report a primer designing graphic user interface program, Pickprimer, that includes gene structure prediction and primer design modules by combining source codes of the Spidey and Primer3 programs. This program provides simple graphic user interface to input sequences and design primers. Genomic sequence and mRNA or coding sequence of genes can be copy and pasted or input as fasta or text files. Based on alignment of the input sequences using the Spidey module, a putative gene structure is graphically visualized along with exon-intron sequences of color codes. Primer design can be easily performed by dragging mouse on the displayed sequences or input primer targeting position with desirable values of primers. The output of designed primers with detailed information is provided by the Primer3 module. PCR evaluation of 24 selected primer sets successfully amplified single amplicons from six Brassica rapa cultivars. The Pickprimer will be a convenient tool for genetic and molecular breeding studies of plants.

New Degenerate Primer for the Cyanobacterial Non-ribosomal Peptides (시아노박테리아 Non-ribosomal Peptides의 효과적인 연구를 위한 New Degenerate Primer의 개발)

  • Kim, Gi-Eun
    • KSBB Journal
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    • v.22 no.5
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    • pp.362-365
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    • 2007
  • Cyanobacteria have been identified as one of the most promising group producing novel biochemically active natural products. Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Large multienzyme complexes which are responsible for the non-ribosomal biosynthesis of peptides are modular for the addition of a single amino acid. An activation of amino acid substrates results in an amino adenylate occuring via an adenylation domain (A-domain). A-domains are responsible for the recognition of amino acids as substrates within NP synthesis. The A-domain contains ten conserved motifs, A1 to A10. In this study, ten conserved motifs from A1 to A10 were checked regarding their amino acid sequence of the NRPS-module of Microcystis aeruginosa PCC 7806. The part of the amino acid sequence chosen was that which contained as many conserved motives as possible, and then these amino sequence were compared between other cyanobacteria to design a new degenerate primer. A new degenerate primer (A3/A7 primer) was designed to detect any putative NP synthetase region in unkwon cyanobacteria by a reverse translation of the conserved amino acid sequence and a search for cyanobacterial DNA bank.

Gene Cloning, Purification and Characterization of Xylanase 10A from Paenibacillus woosongensis in Escherichia coli (Paenibacillus woosongensis로부터 대장균에 Xylanase 10A의 유전자 클로닝과 정제 및 특성분석)

  • Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.158-166
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    • 2020
  • A gene coding for the xylanase was cloned from Paenibacillus woosongensis, followed by determination of its complete nucleotide sequence. This xylanase gene, designated as xyn10A, consists of 1,446 nucleotides encoding a polypeptide of 481 amino acid residues. Based on the deduced amino acid sequence, Xyn10A was identified to be a modular enzyme composed of a catalytic domain highly homologous to the glycosyl hydrolase family 10 xylanase and a putative carbohydrate-binding module (CBM) in the C-terminus. By using DEAE-sepharose and phenyl-sepharose column chromatography, Xyn10A was purified from the cellfree extract of recombinant Escherichia coli carrying a P. woosongensis xyn10A gene. The N-terminal amino acid sequence of the purified Xyn10A was identified to exactly match the sequence immediately following the signal peptide predicted by the Signal5.0 server. The purified Xyn10A was a truncated protein of 33 kDa, suggesting the deletion of CBM in the C-terminus by intracellular hydrolysis. The purified enzyme had an optimum pH and temperature of 6.0 and 55-60℃, respectively, with the kinetic parameters Vmax and Km of 298.8 U/mg and 2.47 mg/ml, respectively, for oat spelt xylan. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchood xylan with low activity for p-nitrophenyl-β-xylopyranoside. Xylanase activity was significantly inhibited by 5 mM Cu2+, Mn2+, and SDS, and was noticeably enhanced by K+, Ni2+, and Ca2+. The enzyme could hydrolyze xylooligosaccharides larger than xylobiose. The predominant products resulting from xylooligosaccharide hydrolysis were xylobiose and xylose.

Development of a Height Adjustment Method of Prefabricated Individual Footing for the Leveling of Unit Modular Structural System (유닛모듈러 건축구조물의 수평 정밀도 확보를 위한 Pre-Fab 독립기초 높이조절 공법 개발)

  • Jun, Young-hun;Kim, Kyoon-Tai;Chae, Myung-Jin
    • Journal of the Korea Institute of Building Construction
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    • v.15 no.6
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    • pp.631-639
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    • 2015
  • In the unit modular construction, the unit module prefabricated in a factory with a high level of accuracy does not fit completely onto the ground-joint junction due to the low accuracy of the ground work. This difference in the level of accuracy can cause diverse problems, such as twisting the upper unit modules and loosening the connection between the module and the footing. On this background, the aim of this study is to develop a technique for adjusting the height of the prefabricated individual footing. To accomplish the aim, a height adjustment method using bolts and nuts is proposed, and a shop drawing and the construction sequence are also presented in this study. The structural safety is verified through a structure simulation. In the future, research will be conducted on a mock-up test of the height adjustment method developed in this study, and an analysis of economic feasibility will be performed in order to verify its constructability and usability.

A Novel pH-Stable, Bifunctional Xylanase Isolated from a Deep-Sea Microorganism, Demequina sp. JK4

  • Meng, Xin;Shao, Zongze;Hong, Yuzhi;Lin, Ling;Li, Chanjuan;Liu, Ziduo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1077-1084
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    • 2009
  • A genomic library was constructed to clone a xylanase gene (Mxyn10) from Demequina sp. JK4 isolated from a deep sea. Mxyn10 encoded a 471 residue protein with a calculated molecular mass of 49 kDa. This protein showed the highest sequence identity (70%) with the xylanase from Streptomyces lividans. Mxyn10 contains a catalytic domain that belongs to the glycoside hydrolase family 10 (GH10) and a carbohydrate-binding module (CBM) belonging to family 2. The optimum pH and temperature for enzymatic activity were pH 5.5 and $55^{\circ}C$, respectively. Mxyn10 exhibited good pH stability, remaining stable after treatment with buffers ranging from pH 3.5 to 10.0. The protein was not significantly affected by a variety of chemical reagents, including some compounds that usually inhibit the activity of other related enzymes. In addition, Mxyn10 showed activity on cellulose. These properties mark Mxyn10 as a potential enzyme for industrial application and saccharification processes essential for bioethanol production.

Study on the Transmission of Medical Information using Bluetooth Technology (블루투스를 이용한 의료정보 신호의 전송에 관한 연구)

  • 엄정규;김영길
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2001.05a
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    • pp.520-523
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    • 2001
  • In this paper, a system that transmits ECG signals which get from hand baggage ECG is implemented by using Bluetooth technology. Bluetooth technology is a close range wireless communication used wireless frequency 2.4GHZ bandwidth. This technology consumes very small power and provides high reliability also self error correction with high speed frequency hopping. Because every device which uses Bluetooth protocol can communicate each other. These can connect between the system implemented and any devices such as mobile telephone with Bluetooth module, notebook, and the personal mobile device. Also, the paper proposes capability of transmission to the sever of hospital through each type of wireless communication device that acquired medical information signals in mobile medical machine. The system consists of hardware parts with Bluetooth module and host part, and software parts with bluetooth protocol stacks. The host precesses a connection with other device and transmits ECG signals with bluetooth frequency hopping sequence.

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A Study on Progressive Working of Electric Product by the using of Fuzzy Set Theory (퍼지 셋 이론을 이용한 전기제품의 프로그레시브 가공에 관한 연구)

  • Kim, J. H;Kim, Y. M.;Kim, Chul;Choi, J. C.
    • Journal of the Korean Society for Precision Engineering
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    • v.19 no.1
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    • pp.79-92
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    • 2002
  • This paper describes a research work of developing computer-aided design of a product with bending and piercing for progressive working. An approach to the system for progressive working is based on the knowledge-based rules. Knowledge for the system is formulated from plasticity theories, experimental results and the empirical knowledge of field experts. The system has been written in AutoLISP on the AutoCAD with a personal computer and is composed of four main modules, which are input and shape treatment, flat pattern layout, strip layout and die layout modules. The system is designed by considering several factors, such as bending sequences by fuzzy set theory, complexities of blank geometry, punch profiles, and the availability of a press equipment. Strip layout drawing generated in the strip layout module is presented in 3-D graphic farms, including bending sequences and piercing processes with punch profiles divided into for external area. The die layout module carries out die design for each process obtained from the results of the strip layout. Results obtained using the modules enable the manufacturer for progressive working of electric products to be more efficient in this field.