Journal of Microbiology

Doo Hyun Nam

7 Articles

Biosynthesis of 2-amino-3-hydroxycyclopent-2-enone moiety of bafilomycin in Kitasatospora cheerisanensis KCTC2395: Nguyen Phan Kieu Hanh , Jae Yoon Hwang , Hye Ryeung Oh , Geum Jin Kim , Hyukjae Choi , Doo Hyun Nam; J. Microbiol. 2018;56(8):571-578. Published online July 25, 2018; DOI: https://doi.org/10.1007/s12275-018-8267-0

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Bafilomycins produced by Kitasatospora cheerisanensis KCTC- 2395 belong to the 16-membered macrolactone family plecomacrolide antibiotics. Bafilomycin B1 contains 2-amino- 3-hydroxycyclopent-2-enone (C5N), a five membered ring, which gets condensed via an amide linkage to bafilomycin polyketide. To study the biosynthetic pathway of C5N during bafilomycin biosynthesis in K. cheerisanensis KCTC2395, we attempted the functional analysis of two putative genes, encoding 5-aminolevulinic acid synthase (ALAS) and acyl- CoA ligase (ACL). The amplified putative genes for ALAS and ACL were cloned into the E. coli expression vector pET- 32a(+) plasmid, following which the soluble recombinant ALAS and ACL proteins were purified through nickel-affinity column chromatography. Through HPLC analysis of the enzyme reaction mixture, we confirmed the products of putative ALAS and ACL reaction as 5-aminolevulinic acid (5- ALA) and 5-ALA-CoA, respectively. The optimal pH for the putative ALAS reaction was 7.5, and for putative ACL reaction was 7.0, as confirmed by the colorimetric assay. Furthermore, pyridoxal 5􍿁-phosphate (PLP) was found to be an essential cofactor in the putative ALAS reaction, and ATP was a cofactor for the putative ACL catalysis. Finally, we also confirmed that the simultaneous treatment of putative ACL and putative ALAS enzymes resulted in the production of C5N compound from 5-ALA.

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Co-culture of Burkholderia vietnamiensis B418 and Trichoderma harzianum T11-W improved the production of nematicidal secondary metabolites against root-knot nematodes in tomato
Wenzhe Li, Yanqing Jiang, Yuyu Zhang, Jishun Li, Jindong Hu, Yilian Wang, Yanli Wei, Hongmei Li, Zhongjuan Zhao, Kai Yang, Hetong Yang, Yi Zhou, Yuanzheng Wu
Biological Control.2026; 218: 106060. CrossRef
Enhanced bafilomycin A1 production in Streptomyces Griseobrunneus via protoplast fusion combined with high-throughput screening
Fei Sun, Yuan Yuan, Haimin Chen, Zhikai Fang, Yeshi Shi, Hong Jiang, Jian Zhou
World Journal of Microbiology and Biotechnology.2025;[Epub] CrossRef
The Secondary Metabolites from Genus Kitasatospora: A Promising Source for Drug Discovery
Yuanjuan Wei, Guiyang Wang, Yan Li, Maoluo Gan
Chemistry & Biodiversity.2024;[Epub] CrossRef
Elucidation of the Late Steps during Hexacosalactone A Biosynthesis in Streptomyces samsunensis OUCT16-12
He Duan, Fang Wang, Chuchu Zhang, Yujing Dong, Huayue Li, Fei Xiao, Wenli Li, Haruyuki Atomi
Applied and Environmental Microbiology.2023;[Epub] CrossRef
A Secondary Metabolic Enzyme Functioned as an Evolutionary Seed of a Primary Metabolic Enzyme
Jun Kawaguchi, Hikaru Mori, Noritaka Iwai, Masaaki Wachi, Miriam Barlow
Molecular Biology and Evolution.2022;[Epub] CrossRef
Biosynthesis of Methoxymalonyl-acyl Carrier Protein (ACP) as an Extender Unit for Bafilomycin Polyketide in Streptomyces griseus DSM 2608
Nguyen Phan Kieu Hanh, Jae Yoon Hwang, Doo Hyun Nam
Biotechnology and Bioprocess Engineering.2018; 23(6): 693. CrossRef

Analysis of a draft genome sequence of Kitasatospora cheerisanensis KCTC 2395 producing bafilomycin antibiotics: Jae Yoon Hwang , Soo Hee Kim , Hye Ryeung Oh , Eunju Kwon , Doo Hyun Nam; J. Microbiol. 2015;53(1):84-89. Published online December 4, 2014; DOI: https://doi.org/10.1007/s12275-015-4340-0

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Kitasatospora cheerisanensis KCTC 2395, producing bafilomycin antibiotics belonging to plecomacrolide group, was isolated from a soil sample at Mt. Jiri, Korea. The draft genome sequence contains 8.04 Mb with 73.6% G+C content and 7,810 open reading frames. All the genes for aerial mycelium and spore formations were confirmed in this draft genome. In phylogenetic analysis of MurE proteins (UDPN- acetylmuramyl-L-alanyl-D-glutamate:DAP ligase) in a conserved dcw (division of cell wall) locus, MurE proteins of Kitasatospora species were placed in a separate clade between MurEs of Streptomyces species incorporating LL-diaminopimelic acid (DAP) and MurEs of Saccharopolyspora erythraea as well as Mycobacterium tuberculosis ligating meso- DAP. From this finding, it was assumed that Kitasatospora MurEs exhibit the substrate specificity for both LL-DAP and meso-DAP. The bafilomycin biosynthetic gene cluster was located in the left subtelomeric region. In 71.3 kb-long gene cluster, 17 genes probably involved in the biosynthesis of bafilomycin derivatives were deduced, including 5 polyketide synthase (PKS) genes comprised of 12 PKS modules.

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Genome-directed discovery of antiproliferative bafilomycins from a deepsea-derived Streptomyces samsunensis
Runyi Wang, Yilei Bao, Yujing Dong, Yun Dong, Huayue Li
Bioorganic Chemistry.2023; 138: 106599. CrossRef
Engineering Bafilomycin High-Producers by Manipulating Regulatory and Biosynthetic Genes in the Marine Bacterium Streptomyces lohii
Zhong Li, Shuai Li, Lei Du, Xingwang Zhang, Yuanyuan Jiang, Wenhua Liu, Wei Zhang, Shengying Li
Marine Drugs.2021; 19(1): 29. CrossRef
Genome based characterization of Kitasatospora sp. MMS16-BH015, a multiple heavy metal resistant soil actinobacterium with high antimicrobial potential
Bo-Ram Yun, Adeel Malik, Seung Bum Kim
Gene.2020; 733: 144379. CrossRef
Genome Mining of the Genus Streptacidiphilus for Biosynthetic and Biodegradation Potential
Adeel Malik, Yu Ri Kim, Seung Bum Kim
Genes.2020; 11(10): 1166. CrossRef
Biosynthesis of 2-amino-3-hydroxycyclopent-2-enone moiety of bafilomycin in Kitasatospora cheerisanensis KCTC2395
Nguyen Phan Kieu Hanh, Jae Yoon Hwang, Hye Ryeung Oh, Geum Jin Kim, Hyukjae Choi, Doo Hyun Nam
Journal of Microbiology.2018; 56(8): 571. CrossRef
Genus Kitasatospora, taxonomic features and diversity of secondary metabolites
Yōko Takahashi
The Journal of Antibiotics.2017; 70(5): 506. CrossRef
Evolution of cyclizing 5-aminolevulinate synthases in the biosynthesis of actinomycete secondary metabolites: outcomes for genetic screening techniques
Kateřina Petříčková, Alica Chroňáková, Tomáš Zelenka, Tomáš Chrudimský, Stanislav Pospíšil, Miroslav Petříček, Václav Krištůfek
Frontiers in Microbiology.2015;[Epub] CrossRef

Biochemical Characteristization of Propionyl-Coenzyme A Carboxylase Complex of Streptomyces toxytricini: Atanas V. Demirev , Anamika Khanal , Nguyen Phan Kieu Hanh , Kyung Tae Nam , Doo Hyun Nam; J. Microbiol. 2011;49(3):407-412. Published online June 30, 2011; DOI: https://doi.org/10.1007/s12275-011-1122-1

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Acyl-CoA carboxylases (ACC) are involved in important primary or secondary metabolic pathways such as fatty acid and/or polyketides synthesis. In the 6.2 kb fragment of pccB gene locus of Streptomyces toxytricini producing a pancreatic inhibitor lipstatin, 3 distinct subunit genes of presumable propionyl-CoA carboxylase (PCCase) complex, assumed to be one of ACC responsible for the secondary metabolism, were identified along with gene for a biotin protein ligase (Bpl). The subunits of PCCase complex were α subunit (AccA3), β subunit (PccB), and auxiliary ε subunit (PccE). In order to disclose the involvement of the PCCase complex in secondary metabolism, some biochemical characteristics of each subunit as well as their complex were examined. In the test of substrate specificity of the PCCase complex, it was confirmed that this complex showed much higher conversion of propionyl-CoA rather than acetyl-CoA. It implies the enzyme complex could play a main role in the production of methylmalonyl-CoA from propionyl-CoA, which is a precursor of secondary polyketide biosynthesis.

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A Novel β/ε Subunit Combination Expands the Tri-Subunit Acyl-CoA Carboxylase Repertoire in Streptomyces coelicolor
Shiyu Wu, Xue Yu, Yujie Wu, Xiaomin Niu, Ximing Chen, Tuo Chen, Wei Zhang, Guangxiu Liu, Paul Dyson
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The degradation of marine abundant compatible solute dimethylsulfoniopropionate was controlled by TetR-family transcriptional regulator DdaR in Alcaligenes faecalis
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A Permissive Medium Chain Acyl-CoA Carboxylase Enables the Efficient Biosynthesis of Extender Units for Engineering Polyketide Carbon Scaffolds
Jun Zhang, Mengmeng Zheng, Jiayan Yan, Zixin Deng, Dongqing Zhu, Xudong Qu
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Pan Wang, Xin Wang, Ying Yin, Mingliang He, Wei Tan, Wenting Gao, Jianping Wen
Frontiers in Microbiology.2021;[Epub] CrossRef
AccR, a TetR Family Transcriptional Repressor, Coordinates Short-Chain Acyl Coenzyme A Homeostasis in Streptomyces avermitilis
Mengya Lyu, Yaqing Cheng, Xiao Han, Ying Wen, Yuan Song, Jilun Li, Zhi Chen, Andrew J. McBain
Applied and Environmental Microbiology.2020;[Epub] CrossRef
Current trends and future prospects of lipstatin: a lipase inhibitor and pro-drug for obesity
Punit Kumar, Kashyap Kumar Dubey
RSC Advances.2015; 5(106): 86954. CrossRef
Biosynthesis of Phenylnannolone A, a Multidrug Resistance Reversal Agent from the Halotolerant Myxobacterium Nannocystis pusilla B150
Sarah M. Bouhired, Max Crüsemann, Celso Almeida, Tilmann Weber, Jörn Piel, Till F. Schäberle, Gabriele M. König
ChemBioChem.2014; 15(5): 757. CrossRef
The TetR Family of Regulators
Leslie Cuthbertson, Justin R. Nodwell
Microbiology and Molecular Biology Reviews.2013; 77(3): 440. CrossRef
Structure and function of biotin-dependent carboxylases
Liang Tong
Cellular and Molecular Life Sciences.2013; 70(5): 863. CrossRef

Identification and Characterization of Acetyl-CoA Carboxylase Gene Cluster in Streptomyces toxytricini: Atanas V. Demirev , Ji Seon Lee , Bhishma R. Sedai , Ivan G. Ivanov , Doo Hyun Nam; J. Microbiol. 2009;47(4):473-478. Published online September 9, 2009; DOI: https://doi.org/10.1007/s12275-009-0135-5

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The gene locus for acetyl-CoA carboxylase (ACC) involved in the primary metabolism was identified from the genomic library of Streptomyces toxytricini which produces a lipase inhibitor lipstatin. The 7.4 kb cloned gene was comprised of 5 ORFs including accD1, accA1, hmgL, fadST1, and stsF. In order to confirm the biochemical characteristics of AccA1, the gene was overexpressed in Escherichia coli cells, and the recombinant protein was purified through Ni2+ affinity chromatography. Because most of the expressed AccA1 was biotinylated by host E. coli BirA in the presence of D-biotin, the non-biotinylated apo-AccA1 was purified after gene induction without D-biotin, followed by exclusion of holo-AccA1 using streptavidin beads. The separated apo-AccA1 was post-translationally biotinylated by S. toxytricini biotin apo-protein ligase (BPL) in a time- and enzyme-dependent manner. This result supports that this gene cluster of S. toxytricini encodes the functional ACC enzyme subunits to be biotinylated.

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Daniel Bär, Benjamin Konetschny, Andreas Kulik, Houchao Xu, Davide Paccagnella, Patrick Beller, Nadine Ziemert, Jeroen S. Dickschat, Bertolt Gust
Microbial Cell Factories.2022;[Epub] CrossRef
Biotechnological and industrial applications of Streptomyces metabolites
Khushboo, Punit Kumar, Kashyap K. Dubey, Zeba Usmani, Minaxi Sharma, Vijai Kumar Gupta
Biofuels, Bioproducts and Biorefining.2022; 16(1): 244. CrossRef
Development and Optimization of a High-Throughput Screening Assay for Rapid Evaluation of Lipstatin Production by Streptomyces Strains
Michal Híreš, Nora Rapavá, Martin Šimkovič, Ľudovít Varečka, Dušan Berkeš, Svetlana Kryštofová
Current Microbiology.2018; 75(5): 580. CrossRef
De novo biosynthesis of myricetin, kaempferol and quercetin in Streptomyces albus and Streptomyces coelicolor
Laura Marín, Ignacio Gutiérrez-del-Río, Rodrigo Entrialgo-Cadierno, Claudio J. Villar, Felipe Lombó, Marie-Joelle Virolle
PLOS ONE.2018; 13(11): e0207278. CrossRef
Regulation and structure of the heteromeric acetyl-CoA carboxylase
Matthew J. Salie, Jay J. Thelen
Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids.2016; 1861(9): 1207. CrossRef
Current trends and future prospects of lipstatin: a lipase inhibitor and pro-drug for obesity
Punit Kumar, Kashyap Kumar Dubey
RSC Advances.2015; 5(106): 86954. CrossRef
Enhanced production of lipstatin from Streptomyces toxytricini by optimizing fermentation conditions and medium
Tingheng Zhu, Lingfei Wang, Weixia Wang, Zhongce Hu, Meilan Yu, Kun Wang, Zhifeng Cui
The Journal of General and Applied Microbiology.2014; 60(3): 106. CrossRef
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Fernando Santos-Beneit, Lorena T Fernández-Martínez, Antonio Rodríguez-García, Seomara Martín-Martín, María Ordóñez-Robles, Paula Yagüe, Angel Manteca, Juan F Martín
Future Microbiology.2014; 9(5): 603. CrossRef
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Sushma Anand, Vijay Singh, Appu Kumar Singh, Monica Mittal, Manish Datt, Bala Subramani, Sangaralingam Kumaran
The FEBS Journal.2012; 279(17): 3214. CrossRef
Biochemical characteristization of propionyl-coenzyme a carboxylase complex of Streptomyces toxytricini
Atanas V. Demirev, Anamika Khanal, Nguyen Phan Kieu Hanh, Kyung Tae Nam, Doo Hyun Nam
The Journal of Microbiology.2011; 49(3): 407. CrossRef
The role of acyl-coenzyme A carboxylase complex in lipstatin biosynthesis of Streptomyces toxytricini
Atanas V. Demirev, Anamika Khanal, Bhishma R. Sedai, Si Kyu Lim, Min Kyun Na, Doo Hyun Nam
Applied Microbiology and Biotechnology.2010; 87(3): 1129. CrossRef

Molecular Cloning of the Phospholipase D Gene from Streptomyces sp. YU100 and Its Expression in Escherichia coli: Ji-Seon Lee , Munkhtsetseg Bat-Ochir , Atanas V. Demirev , Doo Hyun Nam; J. Microbiol. 2009;47(1):116-122. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0161-8

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The gene for phospholipase D (PLD) of Streptomyces sp. YU100 was cloned from λ phage library and heterologously expressed in Escherichia coli. Using an amplified gene fragment based on the consensus sequences of streptomycetes PLDs, λ phage library of Streptomyces sp. YU100 chromosomal DNA was screened. The sequencing result of BamHI-digested 3.8 kb fragment in a positive phage clone revealed the presence of an open reading frame of a full sequence of PLD gene encoding a 540-amino acid protein including 33-amino acid signal peptide. The deduced amino acid sequence showed a high homology with other Streptomyces PLDs, having the highly conserved ‘HKD’ motifs. The PLD gene excluding signal peptide sequence was amplified and subcloned into a pET-32b(+) expression vector in E. coli BL21(DE3). The recombinant PLD was purified by nickel affinity chromatography and compared the enzyme activity with wild-type PLD. The results imply that the recombinant PLD produced by E. coli had the nearly same enzyme activity as PLD from Streptomyces sp. YU100.

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International Journal of Molecular Sciences.2021; 22(19): 10580. CrossRef
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Jing Wang, Sheng Xu, Yang Pang, Xin Wang, Kequan Chen, Pingkai Ouyang
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Zhenxia Zhang, Ming Chen, Wei Xu, Wenli Zhang, Tao Zhang, Cuie Guang, Wanmeng Mu
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Xinyi Tao, Ming Zhao, Ying Zhang, Min Liu, Qinghai Liu, Wei Wang, Feng-Qing Wang, Dongzhi Wei
FEMS Microbiology Letters.2019;[Epub] CrossRef
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Wen-Bin Zhou, Jin-Song Gong, Hai-Juan Hou, Heng Li, Zhen-Ming Lu, Hong-Yu Xu, Zheng-Hong Xu, Jin-Song Shi
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Sai-Sai Li, Yan Li, Neng-Bing Long, Feng Jiang, Rui-Feng Zhang
International Journal of Biological Macromolecules.2018; 117: 1188. CrossRef
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Saovanee Choojit, Uwe T. Bornscheuer, Apichat Upaichit, Aran H‐Kittikun
European Journal of Lipid Science and Technology.2016; 118(5): 803. CrossRef
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Jonathan D. Kornspan, Shlomo Rottem, Ran Nir-Paz, K. C. Carroll
Journal of Clinical Microbiology.2014; 52(5): 1622. CrossRef
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Jonathan D. Kornspan, Shlomo Rottem
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Paige E. Selvy, Robert R. Lavieri, Craig W. Lindsley, H. Alex Brown
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Expression and Biochemical Characteristics of a Phospholipase D from Bacillus licheniformis
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Cloning and Analysis of a Type II Polyketide Synthase Gene Cluster from Streptomyces toxytricini NRRL 15,443: Anna Yoo , Atanas V. Demirev , Ji Seon Lee , Sang Dal Kim , Doo Hyun Nam; J. Microbiol. 2006;44(6):649-654.; DOI: https://doi.org/2462 [pii]

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Abstract PDF: A standard type II polyketide synthase (PKS) gene cluster was isolated while attempting to clone the biosynthetic gene for lipstatin from Streptomyces toxytricini NRRL 15,443. This result was observed using a Southern blot of a PstI-digested S. toxytricini chromosomal DNA library with a 444 bp amplified probe of a ketosynthase (KS) gene fragment. Four open reading frames [thioesterase (TE), β-ketoacyl systhase (KAS), chain length factor (CLF), and acyl carrier protein (ACP)], were identified through the nucleotide sequence determination and analysis of a 4.5 kb cloned DNA fragment. In order to confirm the involvement of a cloned gene in lipstatin biosynthesis, a gene disruption experiment for the KS gene was performed. However, the resulting gene disruptant did not show any significant difference in lipstatin production when compared to wild-type S. toxytricini. This result suggests that lipstatin may not be synthesized by a type II PKS.

Identification of Medicinal Mushroom Species Based on Nuclear Large Subunit rDNA Sequences: Ji Seon Lee , Mi Ok Lim , Kyoung Yeh Cho , Jung Hee Cho , Seung Yeup Chang , Doo Hyun Nam; J. Microbiol. 2006;44(1):29-34.; DOI: https://doi.org/2340 [pii]

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Abstract PDF: The purpose of this study was to develop molecular identification method for medical mushrooms and their preparations based on the nucleotide sequences of nuclear large subunit (LSU) rDNA. Four specimens were collected of each of the three representative medicinal mushrooms used in Korea: Ganoderma lucidum, Coriolus versicolor, and Fomes fomentarius. Fungal material used in these experiments included two different mycelial cultures and two different fruiting bodies from wild or cultivated mushrooms. The genomic DNA of mushrooms were extracted and 3 nuclear LSU rDNA fragments were amplified: set 1 for the 1.1-kb DNA fragment in the upstream region, set 2 for the 1.2-kb fragment in the middle, and set 3 for the 1.3-kb fragment downstream. The amplified gene products of nuclear large subunit rDNA from 3 different mushrooms were cloned into E. coli vector and subjected to nucleotide sequence determination. The sequence thus determined revealed that the gene sequences of the same medicinal mushroom species were more than 99.48% homologous, and the consensus sequences of 3 different medicinal mushrooms were more than 97.80% homologous. Restriction analysis revealed no useful restriction sites for 6-bp recognition enzymes for distinguishing the 3 sequences from one another, but some distinctive restriction patterns were recognized by the 4-bp recognition enzymes AccII and HhaI. This analysis was also confirmed by PCR-RFLP experiments on medicinal mushrooms.

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