• Title/Summary/Keyword: metabolic pathway

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Site-speci fic Inactivation o meso-Diaminopimelate-dehydrogenase Gene (ddh) in a Lysine-producing Brevibacterium lactofementum. (Brevibacterium lactofermentum 에서 meso-Diaminopimelate-dehydrogenase Gene (ddh)의 Site-specific Inactivation)

  • 김옥미;박선희;이갑랑
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.387-392
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    • 1998
  • Brevibacterium lactofermentum, a gram-positive bacteria, has both the diaminopimelate (DAP) pathway and meso-DAP-dehydrogenase (DDH) pathway for L-lysine biosynthesis. To investigate importance of DDH pathway and the related ddh gene in lysine production, we introduced site-specific mutagenesis technique. A 300 bp DNA fragment central to the meso-DAP-dehydrogenase gene (ddh) of B. lactofermentum was used to inactive chromosomal ddh gene via homologous recombination. Southern hybridization analysis confirmed that the chromosomal ddh gene was disrupted by the vector sequence. The B. lactofementum ddh mutant obtained have an inactive DDH pathway. The results reveal that inactivation of the ddh gene in B. lactofermentum leads to dramatic reduction of lysine production as well as decrease of the growth rate, indicating that the DDH pathway is essential for high-level lysine production as well as biosynthesis of meso-DAP.

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CRISPR-Driven Genome Engineering for Chorismate- and Anthranilate-Accumulating Corynebacterium Cell Factories

  • Hye-Jin Kim;Si-Sun Choi;Eung-Soo Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1370-1375
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    • 2023
  • In this study, we aimed to enhance the accumulation of chorismate (CHR) and anthranilate (ANT), key intermediates in the shikimate pathway, by modifying a shikimate over-producing recombinant strain of Corynebacterium glutamicum [19]. To achieve this, we utilized a CRISPR-driven genome engineering approach to compensate for the deletion of shikimate kinase (AroK) as well as ANT synthases (TrpEG) and ANT phosphoribosyltransferase (TrpD). In addition, we inhibited the CHR metabolic pathway to induce CHR accumulation. Further, to optimize the shikimate pathway, we overexpressed feedback inhibition-resistant Escherichia coli AroG and AroH genes, as well as C. glutamicum AroF and AroB genes. We also overexpressed QsuC and substituted shikimate dehydrogenase (AroE). In parallel, we optimized the carbon metabolism pathway by deleting the gntR family transcriptional regulator (IolR) and overexpressing polyphosphate/ATP-dependent glucokinase (PpgK) and glucose kinase (Glk). Moreover, acetate kinase (Ack) and phosphotransacetylase (Pta) were eliminated. Through our CRISPR-driven genome re-design approach, we successfully generated C. glutamicum cell factories capable of producing up to 0.48 g/l and 0.9 g/l of CHR and ANT in 1.3 ml miniature culture systems, respectively. These findings highlight the efficacy of our rational cell factory design strategy in C. glutamicum, which provides a robust platform technology for developing high-producing strains that synthesize valuable aromatic compounds, particularly those derived from the shikimate pathway metabolites.

A Computer-aided Design Tool with Semiautomatic Image-Processing Features for Visualizing Biological Pathways

  • Ham, Sung-Il;Yang, San-Duk;Thong, Chin-Ting;Park, Hyun-Seok
    • Genomics & Informatics
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    • v.7 no.3
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    • pp.168-170
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    • 2009
  • The explosion in biological data resulting from high-throughput experiments requires new software tools to manipulate and display pathways in a way that can integrate disparate sources of information. A visual Java-based CAD tool for drawing and annotating biological pathways with semiautomatic image-processing features is described in this paper. The result of the image-editing process is an XML file for the appropriate links. This tool integrates the pathway images and XML file sources. The system has facilities for linking graphical objects to external databases and is capable of reproducing existing visual representations of pathway maps.

Characterization of Aromatic Hydrocarbon Degrading Bacteria Isolated from Pine Litter (솔잎 퇴적물에서 추출한 방향족 탄화수소물질 분해 박테리아의 동정)

  • Song, Yoon-Jae
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.333-339
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    • 2009
  • Using a novel pine needle agar, fifteen bacterial species were isolated from pine litter. These bacteria were able to degrade aromatic hydrocarbons derived from lignin and utilize the ortho-cleavage of the $\beta$-ketoadipate pathway to degrade protocatechuate or catechol. A different utilization array of aromatic hydrocarbons by these bacteria was also determined. This study provides the information on bacterial species living in pine litter and suggests that these bacteria have metabolic abilities to utilize aromatic hydrocarbons derived from lignin biodegradation.

Functions and Metabolic Pathway of Ascorbic Acid in Plant (식물의 Ascorbic Acid의 기능과 대사령로)

  • Park Yang-Ho;Lee Ju-Young;Jang Byoung-Choon;Lee Ki-Sang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.6
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    • pp.457-461
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    • 2005
  • During the last few years, considerable progresses have been made in understanding of roles and biosynthesis of ascorbic acid (AsA) in plants. The concentrations of ascorbic acid is 2-4 mM in leaf cells, but much higher at the chloroplast. There are three forms of ascorbic acid in the plant mainly ascorbic acid (AsA), monode­hydroascorbic acid (MDHA) and dehydroascorbic acid (DHA). AsA in plant cell performs antioxidants by changing those three forms. And AsA promotes cell division and elongation There was new pathway of ascorbic acid metabolism. It is called pathway of Smirnoff-Wheeler. This report will provide understanding of AsA in plants, and also provide

The Central Concept for Chitin Catabolic Cascade in Marine Bacterium, Vibrios

  • Jung, Byung-Ok;Roseman, Saul;Park, Jae-Kweon
    • Macromolecular Research
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    • v.16 no.1
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    • pp.1-5
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    • 2008
  • The enzymatic hydrolysis of chitin has been studied for almost a century, and early work established that at least two enzymes are required, a chitinase that mainly yields the disaccharide N,N'-diacetylchitobiose, or $(GlcNAc)_2$, and a "chitobiase", or ${\beta}$-N-acetylglucosaminidase, which gives the final product G1cNAc. This pathway has not been completely identified but has remained the central concept for the chitin catabolism through the $20^{th}$ century1 including in marine bacteria. However, the chitin catabolic cascade is quite complex, as described in this review. This report describes three biologically functional genes involved in the chitin catabolic cascade of Vibrios in an attempt to better understand the metabolic pathway of chitin.

Examination of the Central Metabolic Pathway With Genomics in Lactiplantibacillus plantarum K9 (Lactiplantibacillus plantarum K9 유전체 분석을 통해 필수 물질대사 경로의 탐색)

  • Sam Woong Kim;Young Jin Kim;Hyo In Choi;Sang Won Lee;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
    • Journal of Life Science
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    • v.34 no.7
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    • pp.465-475
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    • 2024
  • Lactiplantibacillus plantarum K9 is a probiotic strain that can be utilized from various bioactive substances isolated from Protaetia brevitarsis seulensis larvae. In this study, a genetic analysis of L. plantarum K9 revealed the existence of a bacterial chromosome and three plasmids. The glycolysis pathway and pentose phosphate pathway were examined for their normal functioning via an analysis of the core metabolic pathways of L. plantarum K9. Since the key enzymes, fluctose-1,6-bisphospatase (EC: 3.1.3.11) and 6-phosphogluconate dehydratase (EC: 4.2.1.12)/2-keto-deoxy-6-phosphogluconate (KDPG) aldolase (EC: 4.2.1.55), of gluconeogenesis and the ED pathway were not identified from the L. plantarum K9 genome, we suggest that gluconeogenesis and the ED pathway are not performed in L. plantarum K9. Additionally, while some enzymes, related to fumarate and malate biosyntheses, involved in the TCA cycle were identified from L. plantarum K9, the enzymes associated with the remaining TCA cycle were absent, indicating that the TCA cycle cannot proceed. Meanwhile, based on our findings, we propose that the oxidative electron transport system performs class IIB-type (bd-type) electron transfer. In summary, we assert that L. plantarum K9 performs homolactic fermentation, executes gluconeogenesis and the pentose phosphate pathway, and carries out energy metabolism through the class IIB-type oxidative electron transport system. Therefore, we suggest that L. plantarum K9 has relatively high lactic acid production, and that it has excellent antibacterial activity, as a result, compared to other lactic acid bacterial strains. Moreover, we speculate that L. plantarum K9 has an oxidative electron transport capability, indicating that it is highly resistant to oxygen and suggesting that it has fine cultivation characteristics, which collectively make it highly suitable for use as a probiotic.