• Title/Summary/Keyword: ddh gene

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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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Cloning and Sequencing of the ddh Gene involved in the Novel Pathway of Lysine Biosynthesis from Brevibacterium lactofermentum

  • Kim, Ok-Mi;Kim, Hyun-Jeong;Kim, Dal-Sang;Park, Dong-Chul;Lee, Kap-Rang
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.250-256
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    • 1995
  • The ddh gene encoding meso-diaminopimelate (meso-DAP)-dehydrogenase (DDH) in Brevibacterium lactofermentum was isolated by complementation of the Escherichia coli dapD mutation. It was supposed from subcloning experiments and complementation tests that the evidence for DDH activity appeared in about 2.5 kb Xhol fragmented genome. The 2.5 kb Xhol fragment containing the ddh gene was sequenced, and an open reading frame of 960 bp encoding a polypeptide comprising 320 amino acids was found. Computer analysis indicated that the deduced amino acid of the B. lactofermentum ddh gene showed a high homology with that of the Corynebacterium glutamicum ddh gene.

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L-Lysine Production by Amplification of the ddh Gene in a Lysine-producing Brevibacterium lactofementum. (Brevibacterium lactofermentum에서 ddh 유전자의 증폭에 의한 L-Lysine의 생산)

  • 김옥미;박선희;이승언;배준태;김현정;이별나;이갑랑
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.400-405
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    • 1998
  • The ddh gene encoding meso-DAP-dehydrogenase (DDH) involved in the dehydrogenase pathway is essential for high-level lysine production in Brevibacterium lactofermentum. To investigate the effect of the ddh gene amplification on lysine production by B. lactofementum, we constructed two E. coli -B. lactofermentum shuttle vector, pEB1 and pEB2. The recombinant plasmids, pRK1 and pRK2, carrying the ddh gene were introduced into B. lactofermentum by electroporation. The specific activity of DDH by amplification of the ddh gene was increased 7-fold, and also L-Lysine production of B. lactofermentum strains harboring recombinant plasmids were 18∼20% higher than that of the control.

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Influence on Lysine Production by Overexpression of the ddh Gene in a Lysine-producing Brevibacterium lactofermentum (Brevibacterium lactofermentum에서 ddh 유전자의 Overexpression이 $_L-Lysine$ 생산에 미치는 영향)

  • Park, Sun-Hee;Kim, Ok-Mi;Kim, Hyun-Jeong;Bae, Jun-Tae;Chang, Jong-Sun;Lee, Kap-Rang
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.224-230
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    • 1999
  • The ddh gene encoding meso-DAP-dehydrogenase (DDH) involved in the dehydrogenase pathway is essential for high-level lysine production in Brevibacterium lactofermentum. To investigate its influence on lysine production by overexpression of the ddh gene in a lysine-producing B. lactofermentum, recombinant plasmid pRK1 and pRK31 containing the ddh gene of B. lactofermentum were constructed and they were introduced into B. lactofermentum by electroporation. Multiple copies of pRK1 and pRK31 caused 7-fold and 14-fold increase of DDH activity in B. lactofermentum cell extracts, respectively. As determined in shake flask fermentation, lysine production of B. lactofermentum harboring pRK1 or pRK31 was 22% or 19% higher than that of the control, respectively.

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Combining Ability in Mungbean (Vigna radiata (L.) Wilczek) II. Traits Related to Indetemination

  • Srinives, P.;Khattak, G.S.S.;Haq, M.A.;Ashraf, M.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.5
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    • pp.424-427
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    • 2001
  • Combining ability study was carried out on the components of synchronization in maturity and determinate growth habit in mungbean, using 6$\times$6 diallel cross. Both additive and non-additive gene effects were found conditioning the inheritance of days to first flower, days between first pod and 90% pod maturity (DDd1), plant height from first pod stage to 90% pod maturity (DDhl, DDh2, and DDh3). Only non-additive gene action was important in degree of determination from first pod stage to 90% pod maturity (DDd2). While only additive action was important in plant height at first flower. The predominant additive gene action was observed in all traits but non-additive was significant in only DDd$_2$. For synchronization in maturity, determinate growth habit, and their components, the best combiners were NM92, VCl560D, and NM89, whereas the best indeterminate combinations were NM92 $\times$ NM89, NM92 $\times$ VCl560D, and NM92 $\times$ ML-5.

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Description and Genomic Characteristics of Weissella fermenti sp. nov., Isolated from Kimchi

  • Jae Kyeong Lee;Ju Hye Baek;Dong Min Han;Se Hee Lee;So Young Kim;Che Ok Jeon
    • Journal of Microbiology and Biotechnology
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    • v.33 no.11
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    • pp.1448-1456
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    • 2023
  • A Gram-positive, non-motile, and non-spore-forming lactic acid bacterium, designated as BK2T, was isolated from kimchi, a Korean traditional fermented vegetable food, and the taxonomic characteristics of strain BK2T, along with strain LMG 11983, were analyzed. Both strains optimally grew at 30℃, pH 7.0, and 1.0% NaCl. Cells of both strains were heterofermentative and facultatively anaerobic rods, demonstrating negative reactions for catalase and oxidase. Major fatty acids (>10%) identified in both strains were C18:1 ω9c, C16:0, and summed feature 7 (comprising C19:1 ω6c and/or C19:1 ω7c). The genomic DNA G+C contents of both strains were 44.7 mol%. The 16S rRNA gene sequence similarity (99.9%), average nucleotide identity (ANI; 99.9%), and digital DNA-DNA hybridization (dDDH; 99.7%) value between strains BK2T and LMG 11983 indicated that they are different strains of the same species. Strain BK2T was most closely related to Weissella confusa JCM 1093T and Weissella cibaria LMG 17699T, with 100% and 99.4% 16S rRNA gene sequence similarities, respectively. However, based on the ANI and dDDH values (92.3% and 48.1% with W. confusa, and 78.4% and 23.5% with W. cibaria), it was evident that strain BK2T represents a distinct species separate from W. confusa and W. cibaria. Based on phylogenetic, phenotypic, and chemotaxonomic features, strains BK2T and LMG 11983 represent a novel species of the genus Weissella, for which the name Weissella fermenti sp. nov. is proposed. The type of strain is BK2T (=KACC 22833T=JCM 35750T).

Ramlibacter terrae sp. nov. and Ramlibacter montanisoli sp. nov., Isolated from Soil

  • Khan, Shehzad Abid;Kim, Hyung Min;Baek, Ju Hye;Jung, Hye Su;Jeon, Che Ok
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1210-1217
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    • 2021
  • Two gram-negative, catalase-positive, strictly aerobic, and white colony-forming bacteria, strains H242T and B156T, were isolated from soil in South Korea. Cells of strain H242T were oxidase-positive and non-motile short rods, while those of strain B156T were oxidase-negative and long non-motile rods. Ubiquinone-8 was identified as the sole isoprenoid quinone in both strains. C16:0, cyclo-C17:0, andsummed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol were identified in both strains as the major cellular fatty acids and polar lipids, respectively. The DNA G+C contents of strains H242T and B156T were 69.4 mol% and 69.3 mol%, respectively. Phylogenetic analyses based on 16S rRNA and 92 concatenated core gene sequences revealed that strains H242T and B156T formed distinct phylogenic lineages from other Ramlibacter type strains. The DNA-DNA hybridization (DDH) value between strains H242T and B156T was 24.6%. Strains H242T and B156T were most closely related to Ramlibacter ginsenosidimutans BXN5-27T and Ramlibacter monticola G-3-2T with 98.4% and 98.6% 16S rRNA gene sequence similarities, respectively. Digital DDH values between strain H242T and R. ginsenosidimutans and between strain B156T and R. monticola were 23.5% and 26.1%, respectively. Phenotypic, chemotaxonomic, and molecular analyses indicated that strains H242T and B156T represent two novel species of the genus Ramlibacter, for which the names Ramlibacter terrae sp. nov. and Ramlibacter montanisoli sp. nov., respectively, are proposed. The type strains of R. terrae and R. montanisoli are H242T (=KACC 21667T=JCM 33922T) and B156T (=KACC 21665T=JCM 33920T), respectively.

Paenibacillus gyeongsangnamensis sp. nov., Isolated from Soil

  • Hyosun Lee;Dhiraj Kumar Chaudhary;Dong-Uk Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.8
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    • pp.1636-1641
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    • 2024
  • A Gram-stain-positive, aerobic, white-coloured, rod-shaped bacteria, designated as a strain dW9T, was isolated from soil. Strain dW9T was catalase-positive and oxidase-negative. Strain dW9T grew at temperature of 20-37℃ and at pH of 5.0-7.0. Phylogenetic and 16S rRNA gene analysis indicated that strain dW9T belonged to the genus Paenibacillus with its closest relative being Paenibacillus filicis S4T (97.4% sequence similarity). The genome size of dW9T was 7,787,916 bp with DNA G+C G+C content of 51.3%. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values of dW9T with its closest relatives were found to be <22.0% and <74.0%, respectively. The only respiratory quinone was MK-7, and the major fatty acids were antiso-C15:0 and iso-C16:0. Overall, the comprehensive taxonomic analysis revealed that strain dW9T met all the fundamental criteria to be classified as a novel species within the genus Paenibacillus. Accordingly, we propose the name Paenibacillus gyeongsangnamensis sp. nov., with the type strain dW9T (=KCTC 43431T =NBRC 116022T).

Isolation, Characterization and Whole-Genome Analysis of Paenibacillus andongensis sp.nov. from Korean Soil

  • Yong Guan;Zhun Li;Yoon-Ho Kang;Mi-Kyung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.753-759
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    • 2023
  • The genus Paenibacillus contains a variety of biologically active compounds that have potential applications in a range of fields, including medicine, agriculture, and livestock, playing an important role in the health and economy of society. Our study focused on the bacterium SS4T (KCTC 43402T = GDMCC 1.3498T), which was characterized using a polyphasic taxonomic approach. This strain was analyzed using antiSMASH, BAGEL4, and PRISM to predict the secondary metabolites. Lassopeptide clusters were found using all three analysis methods, with the possibility of secretion. Additionally, PRISM found three biosynthetic gene clusters (BGC) and predicted the structure of the product. Genome analysis indicated that glucoamylase is present in SS4T. 16S rRNA sequence analysis showed that strain SS4T most closely resembled Paenibacillus marchantiophytorum DSM 29850T (98.22%), Paenibacillus nebraskensis JJ-59T (98.19%), and Paenibacillus aceris KCTC 13870T (98.08%). Analysis of the 16S rRNA gene sequences and Type Strain Genome Server (TYGS) analysis revealed that SS4T belongs to the genus Paenibacillus based on the results of the phylogenetic analysis. As a result of the matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) results, SS4T was determined to belong to the genus Paenibacillus. Comparing P. marchantiophytorum DSM 29850T with average nucleotide identity (ANI 78.97%) and digital DNA-DNA hybridization (dDDH 23%) revealed values that were all less than the threshold for bacterial species differentiation. The results of this study suggest that strain SS4T can be classified as a Paenibacillus andongensis species and is a novel member of the genus Paenibacillus.

Caenimonas aquaedulcis sp. nov., Isolated from Freshwater of Daechung Reservoir during Microcystis Bloom

  • Le, Ve Van;Ko, So-Ra;Lee, Sang-Ah;Kang, Mingyeong;Oh, Hee-Mock;Ahn, Chi-Yong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.575-581
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    • 2022
  • A Gram-stain-negative, white-coloured, and rod-shaped bacterium, strain DR4-4T, was isolated from Daechung Reservoir, Republic of Korea, during Microcystis bloom. Strain DR4-4T was most closely related to Caenimonas terrae SGM1-15T and Caenimonas koreensis EMB320T with 98.1% 16S rRNA gene sequence similarities. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain DR4-4T and closely related type strains were below 79.46% and 22.30%, respectively. The genomic DNA G+C content was 67.5%. The major cellular fatty acids (≥10% of the total) were identified as C16:0, cyclo C17:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c), and summed feature 8 (C18:1ω7c and/or C18:1ω6c). Strain DR4-4T possessed phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol as the main polar lipids and Q-8 as the respiratory quinone. The polyamine profile was composed of putrescine, cadaverine, and spermidine. The results of polyphasic characterization indicated that the isolated strain DR4-4T represents a novel species within the genus Caenimonas, for which the name Caenimonas aquaedulcis sp. nov. is proposed. The type strain is DR4-4T (=KCTC 82470T =JCM 34453T).