Ho-Dong Lim - Academia.edu (original) (raw)
Papers by Ho-Dong Lim
Biotechnology and Bioprocess Engineering, Nov 1, 2017
Our aim was to devise targeted drug delivery systems using genetically modified cowpea chlorotic ... more Our aim was to devise targeted drug delivery systems using genetically modified cowpea chlorotic mottle virus (CCMV) capsids by fusion expression with tumorhoming peptide F3 for efficient delivery of therapeutic substances into tumor cells. The RNA-binding domain at the N terminus (amino acid residues 1-25) of CCMV capsid protein (CP) was selectively deleted, and F3 was inserted for the expression in Pichia pastoris. After chromatographic purification, F3-CCMV capsids were obtained via selfassembly of the F3-CP fusion protein and then analyzed by transmission electron microscopy and dynamic light scattering analysis, which revealed spherical nanoparticles (NPs) ca. 18 nm in diameter with regular monodispersity. Near-infrared fluorescent dye IR780 iodide, which has been applied for cancer imaging, photodynamic therapy, and photothermal therapy, was encapsulated in F3-CCMV NPs. The resultant F3-CCMV-IR780 NPs showed excellent molecular targeting to nucleolin receptor overexpressed on the surface of MCF-7 tumor cells. Furthermore, the in vitro cellular uptake and cell viability assay proved a photothermal effect by a single dose of near-infrared laser irradiation. The present system may offer a programmable nanoscaffold-based drug delivery system vehicle for fabrication of promising therapeutic substances for cancer therapy.
한국미생물학회 학술대회논문집, Apr 1, 2014
Journal of Biotechnology, Nov 1, 2010
한국 미생물 생명공학회지, Apr 22, 2022
Biochemical and Biophysical Research Communications, 2018
Bacterial-mediated drug delivery is a potential and promising strategy for the specific treatment... more Bacterial-mediated drug delivery is a potential and promising strategy for the specific treatment of cancer with therapeutic molecules, especially with genetically encoded proteins. These proteins must be tightly regulated due to cytotoxicity and thus are usually expressed under the control of the P BAD and TetA/TetR promoters in vivo. Since protein expression from these systems is triggered by exogenous inducer, periodic intravenous injection of inducer is necessary. However, these treatments can result in non-homogenous and/or inefficient expression of therapeutic proteins in vivo due to impeded diffusion and dilution of the inducer further from the injection site. To overcome these hurdles, we designed a conditional constitutive expression system equipped with the artificial transcription factor, AraC C , which has two operator-binding domains and simultaneously binds to the I 1 and I 2 operators of the P BAD promoter for gene expression in an arabinose-independent manner. Using this construct and the wild type protein AraC under the control of the P BAD promoter, we constructed a self-positive feedback system to constitutively express the therapeutic protein when the induction of AraC was triggered once using arabinose. This expression system could be useful in various cancer treatment strategies using bacteria to deliver genetically encoded drugs in vivo.
KSBB Journal, Mar 31, 2020
한국미생물학회 학술대회논문집, Apr 1, 2014
한국생물공학회 학술대회, Oct 1, 2015
한국생물공학회 학술대회, Apr 1, 2013
International Journal of Molecular Sciences, Jul 22, 2021
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Sustainability, Sep 15, 2020
Manures from livestock industries and farmyards should be managed for land application. Currently... more Manures from livestock industries and farmyards should be managed for land application. Currently, a deep pit or barn system is adopted by many swine farms for manure management, therefore releasing harmful gases and rising the total global emissions of GHGs. This research focuses on the effectiveness of the brown seaweed Sargassum horneri as a masking agent to mitigate odor-generating gaseous pollutants and reduce the emissions of volatile fatty acids (VFAs) from swine manure storage facilities. Using an optimized procedure, we compared the gaseous emissions from two manure storage barns, one containing swine manure masked with S. horneri and the other without masking as a control, over a 30-day period. The results showed that, compared to the control, seaweed masking significantly reduced the sulfide and VFA contents. Furthermore, reductions of 99.48% in H 2 S, 60 ± 5.21% in NH 3 and 74.28 ± 2.14% in gaseous amine emissions were observed within the experimental period. Intriguingly, seaweed masking had beneficial effects, decreasing the total odor content by 97.78 ± 3.15% and increasing the nutrient quality of the manure. S. horneri has great potential as a masking agent in swine manure management to control environmental pollution.
Microorganisms, Jan 8, 2019
Many integrated gene clusters beyond a single genetic element are commonly trapped as the result ... more Many integrated gene clusters beyond a single genetic element are commonly trapped as the result of promoter traps in (meta)genomic DNA libraries. Generally, a single element, which is mainly the promoter, is deduced from the resulting gene clusters and employed to construct a new expression vector. However, expression patterns of target proteins under the incorporated promoter are often inconsistent with those shown in clones harboring plasmids with gene clusters. These results suggest that the integrated set of gene clusters with diverse cis-and transacting elements is evolutionarily tuned as a complete set for gene expression, and is an expression module with all the components for the expression of a nested open reading frame (ORF). This possibility is further supported by truncation and/or serial deletion analysis of this module in which the expression of the nested ORF is highly fluctuated or reduced frequently, despite being supported by plentiful cis-acting elements in the spanning regions around the ORF such as the promoter, ribosome binding site (RBS), terminator, and 3-/5-UTRs for gene expression. Here, we examined whether an innate module with a naturally overexpressed gene could be considered as a scaffold for an expression system. For a proof-of-principle study, we mined a complete expression module with an innately overexpressed ORF in E. coli from a metagenomics DNA library, and incorporated it into a vector that had no regulatory element for expressing the insert. We obtained successful expression of several inserts such as MBP, GFPuv, β-glucosidase, and esterase using this simple construct without tuning and codon optimization of the target insert.
Oncotarget, Jan 5, 2018
Bacterial cancer therapy relies on the properties of certain bacterial species capable of targeti... more Bacterial cancer therapy relies on the properties of certain bacterial species capable of targeting and proliferating within solid malignancies. If these bacteria could be loaded with antitumor proteins, the efficacy of this approach could be greatly increased. However, because most antitumor proteins are also toxic to normal tissue, they must be expressed by bacteria that specifically target and exclusively localize to tumor tissue. As a strategy for treating solid malignancies, we recently evaluated L-asparaginase (L-ASNase) delivered by tumor-targeted Salmonella. In this system, L-ASNase was expressed under the control of the araBAD promoter (PBAD) of the E. coli arabinose operon, which is induced by injection of L-arabinose. Here, we further improved the performance of recombinant Salmonella in cancer therapy by exploiting the quorum-sensing (QS) system, which uses cell mass-dependent auto-induction logic. This approach obviates the necessity of monitoring intratumoral bacterial status and inducing cargo protein expression by administration of an exogenous compound. Recombinant Salmonella in tumors expressed and secreted active L-ASNase in a cell mass-dependent manner, yielding significant anticancer effects. These results suggest that expression of a therapeutic protein under the control of the QS system represents a promising engineering platform for the production of recombinant proteins in vivo.
Antibiotics, Sep 9, 2021
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Journal of Microbiology and Biotechnology, Jul 28, 2017
Frontiers of Environmental Science & Engineering, Mar 25, 2020
Livestock waste is commonly converted into cost-effective and efficient forms of agricultural man... more Livestock waste is commonly converted into cost-effective and efficient forms of agricultural manure. However, in recent years, livestock farming has faced critical challenges
Algal Research-Biomass Biofuels and Bioproducts, Apr 1, 2018
One of the most intensively studied species in red algae is Gracilaria spp., which is also a high... more One of the most intensively studied species in red algae is Gracilaria spp., which is also a highly abundantly produced species annually in Asian countries including Korea and Japan. Much of the recent work on production of reducing sugars from Gracilaria spp. has concentrated on a combined process of physicochemical and biochemical treatments using acids and commercial enzymes. To the best of our knowledge, this study is first demonstration of combined acid and enzymatic hydrolysis using recombinant agarase to produce total reducing sugar (TRS) from Gracilaria verrucosa. By performing acid hydrolysis under the specified condition (2% biomass, 0.1 N HCl, 121°C, 15 min), a 34.9% TRS yield was obtained. Subsequent enzymatic hydrolysis using two recombinant agarases (aga 50D 0.5 U/mg + NABH 0.5 U/mg), resulted in a 47.4% TRS yield was achieved. The TRS yield obtained from acid and enzymatic hydrolysates was 5% higher than that of previous study with commercial enzyme. These results served to demonstrate that feasibility of enzymatic hydrolysis using the recombinant agarases to produce TRS from G. verrucosa for bio-refinery applications.
Microbiology and Biotechnology Letters
Biotechnology and Bioprocess Engineering, Nov 1, 2017
Our aim was to devise targeted drug delivery systems using genetically modified cowpea chlorotic ... more Our aim was to devise targeted drug delivery systems using genetically modified cowpea chlorotic mottle virus (CCMV) capsids by fusion expression with tumorhoming peptide F3 for efficient delivery of therapeutic substances into tumor cells. The RNA-binding domain at the N terminus (amino acid residues 1-25) of CCMV capsid protein (CP) was selectively deleted, and F3 was inserted for the expression in Pichia pastoris. After chromatographic purification, F3-CCMV capsids were obtained via selfassembly of the F3-CP fusion protein and then analyzed by transmission electron microscopy and dynamic light scattering analysis, which revealed spherical nanoparticles (NPs) ca. 18 nm in diameter with regular monodispersity. Near-infrared fluorescent dye IR780 iodide, which has been applied for cancer imaging, photodynamic therapy, and photothermal therapy, was encapsulated in F3-CCMV NPs. The resultant F3-CCMV-IR780 NPs showed excellent molecular targeting to nucleolin receptor overexpressed on the surface of MCF-7 tumor cells. Furthermore, the in vitro cellular uptake and cell viability assay proved a photothermal effect by a single dose of near-infrared laser irradiation. The present system may offer a programmable nanoscaffold-based drug delivery system vehicle for fabrication of promising therapeutic substances for cancer therapy.
한국미생물학회 학술대회논문집, Apr 1, 2014
Journal of Biotechnology, Nov 1, 2010
한국 미생물 생명공학회지, Apr 22, 2022
Biochemical and Biophysical Research Communications, 2018
Bacterial-mediated drug delivery is a potential and promising strategy for the specific treatment... more Bacterial-mediated drug delivery is a potential and promising strategy for the specific treatment of cancer with therapeutic molecules, especially with genetically encoded proteins. These proteins must be tightly regulated due to cytotoxicity and thus are usually expressed under the control of the P BAD and TetA/TetR promoters in vivo. Since protein expression from these systems is triggered by exogenous inducer, periodic intravenous injection of inducer is necessary. However, these treatments can result in non-homogenous and/or inefficient expression of therapeutic proteins in vivo due to impeded diffusion and dilution of the inducer further from the injection site. To overcome these hurdles, we designed a conditional constitutive expression system equipped with the artificial transcription factor, AraC C , which has two operator-binding domains and simultaneously binds to the I 1 and I 2 operators of the P BAD promoter for gene expression in an arabinose-independent manner. Using this construct and the wild type protein AraC under the control of the P BAD promoter, we constructed a self-positive feedback system to constitutively express the therapeutic protein when the induction of AraC was triggered once using arabinose. This expression system could be useful in various cancer treatment strategies using bacteria to deliver genetically encoded drugs in vivo.
KSBB Journal, Mar 31, 2020
한국미생물학회 학술대회논문집, Apr 1, 2014
한국생물공학회 학술대회, Oct 1, 2015
한국생물공학회 학술대회, Apr 1, 2013
International Journal of Molecular Sciences, Jul 22, 2021
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Sustainability, Sep 15, 2020
Manures from livestock industries and farmyards should be managed for land application. Currently... more Manures from livestock industries and farmyards should be managed for land application. Currently, a deep pit or barn system is adopted by many swine farms for manure management, therefore releasing harmful gases and rising the total global emissions of GHGs. This research focuses on the effectiveness of the brown seaweed Sargassum horneri as a masking agent to mitigate odor-generating gaseous pollutants and reduce the emissions of volatile fatty acids (VFAs) from swine manure storage facilities. Using an optimized procedure, we compared the gaseous emissions from two manure storage barns, one containing swine manure masked with S. horneri and the other without masking as a control, over a 30-day period. The results showed that, compared to the control, seaweed masking significantly reduced the sulfide and VFA contents. Furthermore, reductions of 99.48% in H 2 S, 60 ± 5.21% in NH 3 and 74.28 ± 2.14% in gaseous amine emissions were observed within the experimental period. Intriguingly, seaweed masking had beneficial effects, decreasing the total odor content by 97.78 ± 3.15% and increasing the nutrient quality of the manure. S. horneri has great potential as a masking agent in swine manure management to control environmental pollution.
Microorganisms, Jan 8, 2019
Many integrated gene clusters beyond a single genetic element are commonly trapped as the result ... more Many integrated gene clusters beyond a single genetic element are commonly trapped as the result of promoter traps in (meta)genomic DNA libraries. Generally, a single element, which is mainly the promoter, is deduced from the resulting gene clusters and employed to construct a new expression vector. However, expression patterns of target proteins under the incorporated promoter are often inconsistent with those shown in clones harboring plasmids with gene clusters. These results suggest that the integrated set of gene clusters with diverse cis-and transacting elements is evolutionarily tuned as a complete set for gene expression, and is an expression module with all the components for the expression of a nested open reading frame (ORF). This possibility is further supported by truncation and/or serial deletion analysis of this module in which the expression of the nested ORF is highly fluctuated or reduced frequently, despite being supported by plentiful cis-acting elements in the spanning regions around the ORF such as the promoter, ribosome binding site (RBS), terminator, and 3-/5-UTRs for gene expression. Here, we examined whether an innate module with a naturally overexpressed gene could be considered as a scaffold for an expression system. For a proof-of-principle study, we mined a complete expression module with an innately overexpressed ORF in E. coli from a metagenomics DNA library, and incorporated it into a vector that had no regulatory element for expressing the insert. We obtained successful expression of several inserts such as MBP, GFPuv, β-glucosidase, and esterase using this simple construct without tuning and codon optimization of the target insert.
Oncotarget, Jan 5, 2018
Bacterial cancer therapy relies on the properties of certain bacterial species capable of targeti... more Bacterial cancer therapy relies on the properties of certain bacterial species capable of targeting and proliferating within solid malignancies. If these bacteria could be loaded with antitumor proteins, the efficacy of this approach could be greatly increased. However, because most antitumor proteins are also toxic to normal tissue, they must be expressed by bacteria that specifically target and exclusively localize to tumor tissue. As a strategy for treating solid malignancies, we recently evaluated L-asparaginase (L-ASNase) delivered by tumor-targeted Salmonella. In this system, L-ASNase was expressed under the control of the araBAD promoter (PBAD) of the E. coli arabinose operon, which is induced by injection of L-arabinose. Here, we further improved the performance of recombinant Salmonella in cancer therapy by exploiting the quorum-sensing (QS) system, which uses cell mass-dependent auto-induction logic. This approach obviates the necessity of monitoring intratumoral bacterial status and inducing cargo protein expression by administration of an exogenous compound. Recombinant Salmonella in tumors expressed and secreted active L-ASNase in a cell mass-dependent manner, yielding significant anticancer effects. These results suggest that expression of a therapeutic protein under the control of the QS system represents a promising engineering platform for the production of recombinant proteins in vivo.
Antibiotics, Sep 9, 2021
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Journal of Microbiology and Biotechnology, Jul 28, 2017
Frontiers of Environmental Science & Engineering, Mar 25, 2020
Livestock waste is commonly converted into cost-effective and efficient forms of agricultural man... more Livestock waste is commonly converted into cost-effective and efficient forms of agricultural manure. However, in recent years, livestock farming has faced critical challenges
Algal Research-Biomass Biofuels and Bioproducts, Apr 1, 2018
One of the most intensively studied species in red algae is Gracilaria spp., which is also a high... more One of the most intensively studied species in red algae is Gracilaria spp., which is also a highly abundantly produced species annually in Asian countries including Korea and Japan. Much of the recent work on production of reducing sugars from Gracilaria spp. has concentrated on a combined process of physicochemical and biochemical treatments using acids and commercial enzymes. To the best of our knowledge, this study is first demonstration of combined acid and enzymatic hydrolysis using recombinant agarase to produce total reducing sugar (TRS) from Gracilaria verrucosa. By performing acid hydrolysis under the specified condition (2% biomass, 0.1 N HCl, 121°C, 15 min), a 34.9% TRS yield was obtained. Subsequent enzymatic hydrolysis using two recombinant agarases (aga 50D 0.5 U/mg + NABH 0.5 U/mg), resulted in a 47.4% TRS yield was achieved. The TRS yield obtained from acid and enzymatic hydrolysates was 5% higher than that of previous study with commercial enzyme. These results served to demonstrate that feasibility of enzymatic hydrolysis using the recombinant agarases to produce TRS from G. verrucosa for bio-refinery applications.
Microbiology and Biotechnology Letters