• Title/Summary/Keyword: Systems Biotechnology

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Enhanced Large-Scale Production of Hahella chejuensis-Derived Prodigiosin and Evaluation of Its Bioactivity

  • Jeong, Yu-jin;Kim, Hyun Ju;Kim, Suran;Park, Seo-Young;Kim, HyeRan;Jeong, Sekyoo;Lee, Sang Jun;Lee, Moo-Seung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1624-1631
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    • 2021
  • Prodigiosin as a high-valued compound, which is a microbial secondary metabolite, has the potential for antioxidant and anticancer effects. However, the large-scale production of functionally active Hahella chejuensis-derived prodigiosin by fermentation in a cost-effective manner has yet to be achieved. In the present study, we established carbon source-optimized medium conditions, as well as a procedure for producing prodigiosin by fermentation by culturing H. chejuensis using 10 L and 200 L bioreactors. Our results showed that prodigiosin productivity using 250 ml flasks was higher in the presence of glucose than other carbon sources, including mannose, sucrose, galactose, and fructose, and could be scaled up to 10 L and 200 L batches. Productivity in the glucose (2.5 g/l) culture while maintaining the medium at pH 6.89 during 10 days of cultivation in the 200 L bioreactor was measured and increased more than productivity in the basal culture medium in the absence of glucose. Prodigiosin production from 10 L and 200 L fermentation cultures of H. chejuensis was confirmed by high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) analyses for more accurate identification. Finally, the anticancer activity of crude extracted prodigiosin against human cancerous leukemia THP-1 cells was evaluated and confirmed at various concentrations. Conclusively, we demonstrate that culture conditions for H. chejuensis using a bioreactor with various parameters and ethanol-based extraction procedures were optimized to mass-produce the marine bacterium-derived high purity prodigiosin associated with anti-cancer activity.

Ligand Based Pharmacophore Identification and Molecular Docking Studies for Grb2 Inhibitors

  • Arulalapperumal, Venkatesh;Sakkiah, Sugunadevi;Thangapandian, Sundarapandian;Lee, Yun-O;Meganathan, Chandrasekaran;Hwang, Swan;Lee, Keun-Woo
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1707-1714
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    • 2012
  • Grb2 is an adapter protein involved in the signal transduction and cell communication. The Grb2 is responsible for initiation of kinase signaling by Ras activation which leads to the modification in transcription. Ligand based pharmacophore approach was applied to built the suitable pharmacophore model for Grb2. The best pharmacophore model was selected based on the statistical values and then validated by Fischer's randomization method and test set. Hypo1 was selected as a best pharmacophore model based on its statistical values like high cost difference (182.22), lowest RMSD (1.273), and total cost (80.68). It contains four chemical features, one hydrogen bond acceptor (HBA), two hydrophobic (HY), and one ring aromatic (RA). Fischer's randomization results also shows that Hypo1 have a 95% significant level. The correlation coefficient of test set was 0.97 which was close to the training set value (0.94). Thus Hypo1 was used for virtual screening to find the potent inhibitors from various chemical databases. The screened compounds were filtered by Lipinski's rule of five, ADMET and subjected to molecular docking studies. Totally, 11 compounds were selected as a best potent leads from docking studies based on the consensus scoring function and critical interactions with the amino acids in Grb2 active site.

Development of rice(Oryza sativa L.) transformation system to improve callus utilization (캘러스 활용도를 향상시키기 위한 벼(Oryza sativa L.) 형질전환 시스템 구축)

  • Park, Ji-Sun;Moon, Ki-Beom;Ha, Jang-Ho;Jang, Ji-Young;Kim, Mi-Jin;Jeon, Jae-Heung;Park, Sang-Un;Kim, Hyun-Soon
    • Korean Journal of Breeding Science
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    • v.49 no.3
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    • pp.170-179
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    • 2017
  • Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.

Properties of Photo-Reactive Natural Polymer Derivatives and Its Applications (광반응성 천연 고분자의 특성 및 생체재료로의 활용 방안)

  • Kim, Eun-Hye;Jeong, Jin-Hong;Han, Ga-Dug;Son, Tae-Il
    • Prospectives of Industrial Chemistry
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    • v.18 no.4
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    • pp.1-9
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    • 2015
  • 최근 신체의 손상된 조직 및 세포를 재생하기 위해 약물 전달이 가능한 생체재료에 관한 연구가 활발히 이루어지고 있다. 이러한 생체재료 개발을 위해 생체적합하며 생분해성이 뛰어난 천연고분자가 큰 각광을 받고 있다. 기존의 약물 전달을 위한 고정화 방법은 화학적 가교가 널리 이용되어 왔으나, 이 방법의 여러 단점들이 보고된 바 있다. 이러한 단점을 극복하기 위해 광반응성 천연고분자를 이용한 약물광고정화 방법이 연구되어 왔다. 본 글에서는 광고정화를 위해 합성되는 여러 광반응성 천연고분자의 종류 및 특성과 생체재료로써의 활용 방안을 소개하고자 한다.

Comparative Analysis of T4SS Molecular Architectures

  • Mishghan Zehra;Jiwon Heo;Jeong Min Chung;Clarissa L Durie
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1543-1551
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    • 2023
  • The recently published high-resolution R388 T4SS structure provides exciting new details about the complete complex of T4SS, including the components making up the stalk and arches, numerous symmetry mismatches between regions of the complex, and an intriguing interpretation of the closed stalk and radial symmetry of the inner membrane complex, which is related to pilus biogenesis assembly. However, there are a few unidentified densities in the electron microscopy map and portions of the identified component sequences for which the structure is not yet known. It is also unclear how well this minimized DNA-transporting T4SS predicts the structure of other T4SSs, such as expanded systems and those that transport proteins rather than DNA. In this review, we evaluate what can be inferred from the recent high-resolution structure of the R388 T4SS with respect to the Cag and Dot/Icm systems. These systems were selected because, given what is currently known about these systems, we expect them to present most structural differences compared to the R388 T4SS structure. Furthermore, we discuss bacterial physiology and diversity, the T4SS structures and their variations between different bacterial species. These insights may prove beneficial for researchers who elucidate the structure and functions of T4SS in different bacterial species.

Functional pathogenomics of Burkhozderia glumae (oral)

  • Kim, Jinwoo;Kim, Suhyun;Yongsung Kang;Jang, Ji-Youn;Kim, Jung-Gun;Lim, Jae-Yoon;Kim, Minkyun;Ingyu Hwang
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.66.1-66
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    • 2003
  • The aim of this study was to characterize the interactions of rice and Burkholderia glumae, a causal agent of bacterial grain rot of rice, at molecular levels using whole genomic sequences and to identify genes important for pathogenicity and symptom development. To do these, we sequenced whole genome of the bacterium and constructed cosmid clone profiles. We generated pools of mutants using various transposons and determined mutation sites by sequencing rescued plasmids. We focused on studying toxoflavin biosynthetic genes, quorum sensing regulation, and Hrp type III protein secretion systems. We found that two possible operons consisting of five genes are involved in toxoflavin biosynthesis and their expression is regulated by quorum sensing and LysR-type regulator, ToxR. We have isolated the nn PAI of B. glumae and characterized by mutational analyses. The hrp cluster resembled most the putative Type III secretion systems of B. pseudomallei, which is the causative agent of melioidosis, a serious disease of man and animals. The Hrp PAI core region showed high similarity to that of Ralstonia solanacearum and Xanthomonas campestris, however some aspects were dissimilar.

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Effects of Chaperones on mRNA Stability and Gene Expression in Escherichia coli

  • Yoon, Hyun-Jin;Hong, Ji-Young;Ryu, Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.228-233
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    • 2008
  • Effects of chaperones on mRNA stability and gene expression were studied in order to develop an efficient Escherichia coli expression system that can maximize gene expression. The stability of mRNA was modulated by introducing various secondary structures at the 5'-end of mRNA. Four vector systems providing different 5'-end structures were constructed, and genes encoding GFPuv and endoxylanase were cloned into the four vector systems. Primer extension assay revealed different mRNA half-lives depending on the 5'-end secondary structures of mRNA. In addition to the stem-loop structure at the 5'-end of mRNA, coexpression of dnaK-dnaJ-grpE or groEL-groES, representative heat-shock genes in E. coli, increased the mRNA stability and the level of gene expression further, even though the degree of stabilization was varied. Our work suggests that some of the heat-shock proteins can function as mRNA stabilizers as well s protein chaperones.

A Genome-Scale Co-Functional Network of Xanthomonas Genes Can Accurately Reconstruct Regulatory Circuits Controlled by Two-Component Signaling Systems

  • Kim, Hanhae;Joe, Anna;Lee, Muyoung;Yang, Sunmo;Ma, Xiaozhi;Ronald, Pamela C.;Lee, Insuk
    • Molecules and Cells
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    • v.42 no.2
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    • pp.166-174
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    • 2019
  • Bacterial species in the genus Xanthomonas infect virtually all crop plants. Although many genes involved in Xanthomonas virulence have been identified through molecular and cellular studies, the elucidation of virulence-associated regulatory circuits is still far from complete. Functional gene networks have proven useful in generating hypotheses for genetic factors of biological processes in various species. Here, we present a genome-scale co-functional network of Xanthomonas oryze pv. oryzae (Xoo) genes, XooNet (www.inetbio.org/xoonet/), constructed by integrating heterogeneous types of genomics data derived from Xoo and other bacterial species. XooNet contains 106,000 functional links, which cover approximately 83% of the coding genome. XooNet is highly predictive for diverse biological processes in Xoo and can accurately reconstruct cellular pathways regulated by two-component signaling transduction systems (TCS). XooNet will be a useful in silico research platform for genetic dissection of virulence pathways in Xoo.

Characterization of Prototype Foamy Virus Infectivity in Transportin 3 Knockdown Human 293t Cell Line

  • Hamid, Faysal Bin;Kim, Jinsun;Shin, Cha-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.380-387
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    • 2017
  • The foamy viruses are currently considered essential for development as vectors for gene delivery. Previous studies demonstrated that prototype foamy virus (PFV) can infect and replicate prevalently in a variety of cell types for its exclusive replication strategy. However, the virus-host interaction, especially PFV-transportin3 (TNPO3), is still poorly understood. In our investigation of the role of TNPO3 in PFV infection, we found lower virus production in TNPO3 knockdown (KD) cells compared with wild-type 293T cells. PCR analysis revealed that viral DNAs were mostly altered to circular forms: both 1-long terminal repeat (1-LTR) and 2-LTR in TNPO3 KD cells. We therefore suggest that TNPO3 is required for successful PFV replication, at least at/after the nuclear entry step of viral DNA. These findings highlight the obscure mysteries of PFV-host interaction and the requirement of TNPO3 for productive infection of PFV in 293T cells.