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Biotechnological application of Bacillus subtilis spore display system
New Biotechnology · Volume 31S · July 2014
GENERAL BIOTECHNOLOGY
did not inhibit DHT induction of MCP-1 mRNA. Moreover, conditioned media from androgen-treated cells promoted human monocyte THP-1 cell migration and this effect was inhibited by antibody against MCP-1. These results indicate that androgen may regulate MCP-1 and promote inflammatory microenvironment in prostate cancer. http://dx.doi.org/10.1016/j.nbt.2014.05.1982
used CotE, CotG, CotY spore coat protein as anchoring motives for the successful display of beta-galactosidase, spreptavidin, and other interesting target protein with biotechnological application such as laccase for the environmental usages. Surface localization using flow cytometry and enzymatic activity on the Bacillus subtilis spore was examined for the successful confirmation of surface localization of target proteins. Further usage and other application will be discussed. http://dx.doi.org/10.1016/j.nbt.2014.05.1984
PI-14 Suppression of dust mite extract and 2,4dinitrochlorobenzene-induced atopic dermatitis by the water extract of DA-9601 Choi Eun-Ju ∗ , Kim Eun-Kyung Konkuk University
DA-9601 is a novel anti-peptic formulation prepared from the ethanol extracts of Artemisia asiatica possessing anti-oxidative, anti-allergic and anti-inflammatory activities. However, their effect on atopic dermatitis (AD) has not been studied yet. In this study, we report that topical application of DA-9601 suppressed house dust mite extract (Dermatophagoides farinae extract, DFE) and 2, 4-dinitrochlorobenzene (CDNB)-induced AD-like skin lesions in BALB/c mice model. We established atopic dermatitis model in BALB/c mice by repeated local exposure of DFE/CDNB to the ears. Repeated alternative treatment of DFE/CDNB caused ADlike lesions. DA-9601 reduced AD-like skin lesions based on ear thickness and histopathological analysis, and serum IgE levels. DA9601 inhibited mast cell infiltration into the ear and elevation of serum histamine in AD model. In addition, DA-9601 suppressed DFE/CDNB-induced expression of IL-4, IL-13, IL-31, and TNF-␣ in the ears. Taken together, our results showed that topical application of DA-9601 exerts beneficial effects in animal model of AD, suggesting that DA-9601 might be a candidate for the treatment of AD. http://dx.doi.org/10.1016/j.nbt.2014.05.1983
PI-15 Biotechnological application of Bacillus subtilis spore display system June Hyung Kim Dong-A University
Bacterial surface display finds its important biotechnological application in the fields of screening tools of evolved enzyme, bioremediation, whole cell bioconversion and tool for live vaccine production. For the functional bacterial surface display of active enzyme of multimeric form, which is generally impossible due to molecular assembly of the monomer subunit subsequent to the secretion of displayed target protein outside the cell, a new surface display system based on Bacillus subtilis spore is developed. Here, we tried to develop a new bacterial surface display format for the efficient expression of multi-subunit enzyme. We S148
www.elsevier.com/locate/nbt
PI-16 Production of 3-hydroxybutyrate by E. coli: Application of Nitrogen and Phosphorous limitation to steer fluxes to product formation Monica Guevara 1,∗ , Johan Jarmander 1 , Mariel Perez-Zabaleta 1 , Jorge Quillaguamán 2 , Gen Larsson 1 1
Industrial biotechnology, School of biotechnology, KTH, Stockholm Center of Biotechnology, Faculty of Science and Technology, San Simón University, Cochabamba Bolivia
2
Polyhydroxyalcanoates are polyesters produced in large amounts by different microorganisms, but not by E.coli. Here we expressed the bacterial polyhydroxybutarate (PHB) pathway of Halomonas boliviensis in E. coli using phbA and phbB genes to produce the monomer 3-HB. Acetyl coenzyme A (AcCoA) is the intermediate of the carbon metabolism and precursor of the PHB pathway. The objective is to design a process to produce 3-HB in E.coli with high productivity which is a first step to further produce different quality polyesters. Production of the wild type microorganisms takes place under nutrient deficient conditions and excess of a carbon source. The hypothesis is that directing the flux to the desired product (3-HB) by cultivations with controlled feed of nitrogen or phosphorous will increase the flux to the precursor (AcCoA) and minimize the formation of byproducts This will be done by: Fed-Batch cultivations with an excess of glucose and nitrogen limitation. Excess glucose will give a surplus of NADH that will inhibit citrate synthase and the AcCoA produced will instead be used for 3-HB production Fed-Batch cultivations with an excess of glucose and phosphorous limitation. Excess glucose will give a surplus of NADH that will inhibit citrate synthase and the AcCoA produced will instead be used for 3-HB production. Also the limitation of phosphorous activates the methylglyoxal pathway also leading to the more energy favorable production of AcCoA than central glycolisis. http://dx.doi.org/10.1016/j.nbt.2014.05.1985
GENERAL BIOTECHNOLOGY
did not inhibit DHT induction of MCP-1 mRNA. Moreover, conditioned media from androgen-treated cells promoted human monocyte THP-1 cell migration and this effect was inhibited by antibody against MCP-1. These results indicate that androgen may regulate MCP-1 and promote inflammatory microenvironment in prostate cancer. http://dx.doi.org/10.1016/j.nbt.2014.05.1982
used CotE, CotG, CotY spore coat protein as anchoring motives for the successful display of beta-galactosidase, spreptavidin, and other interesting target protein with biotechnological application such as laccase for the environmental usages. Surface localization using flow cytometry and enzymatic activity on the Bacillus subtilis spore was examined for the successful confirmation of surface localization of target proteins. Further usage and other application will be discussed. http://dx.doi.org/10.1016/j.nbt.2014.05.1984
PI-14 Suppression of dust mite extract and 2,4dinitrochlorobenzene-induced atopic dermatitis by the water extract of DA-9601 Choi Eun-Ju ∗ , Kim Eun-Kyung Konkuk University
DA-9601 is a novel anti-peptic formulation prepared from the ethanol extracts of Artemisia asiatica possessing anti-oxidative, anti-allergic and anti-inflammatory activities. However, their effect on atopic dermatitis (AD) has not been studied yet. In this study, we report that topical application of DA-9601 suppressed house dust mite extract (Dermatophagoides farinae extract, DFE) and 2, 4-dinitrochlorobenzene (CDNB)-induced AD-like skin lesions in BALB/c mice model. We established atopic dermatitis model in BALB/c mice by repeated local exposure of DFE/CDNB to the ears. Repeated alternative treatment of DFE/CDNB caused ADlike lesions. DA-9601 reduced AD-like skin lesions based on ear thickness and histopathological analysis, and serum IgE levels. DA9601 inhibited mast cell infiltration into the ear and elevation of serum histamine in AD model. In addition, DA-9601 suppressed DFE/CDNB-induced expression of IL-4, IL-13, IL-31, and TNF-␣ in the ears. Taken together, our results showed that topical application of DA-9601 exerts beneficial effects in animal model of AD, suggesting that DA-9601 might be a candidate for the treatment of AD. http://dx.doi.org/10.1016/j.nbt.2014.05.1983
PI-15 Biotechnological application of Bacillus subtilis spore display system June Hyung Kim Dong-A University
Bacterial surface display finds its important biotechnological application in the fields of screening tools of evolved enzyme, bioremediation, whole cell bioconversion and tool for live vaccine production. For the functional bacterial surface display of active enzyme of multimeric form, which is generally impossible due to molecular assembly of the monomer subunit subsequent to the secretion of displayed target protein outside the cell, a new surface display system based on Bacillus subtilis spore is developed. Here, we tried to develop a new bacterial surface display format for the efficient expression of multi-subunit enzyme. We S148
www.elsevier.com/locate/nbt
PI-16 Production of 3-hydroxybutyrate by E. coli: Application of Nitrogen and Phosphorous limitation to steer fluxes to product formation Monica Guevara 1,∗ , Johan Jarmander 1 , Mariel Perez-Zabaleta 1 , Jorge Quillaguamán 2 , Gen Larsson 1 1
Industrial biotechnology, School of biotechnology, KTH, Stockholm Center of Biotechnology, Faculty of Science and Technology, San Simón University, Cochabamba Bolivia
2
Polyhydroxyalcanoates are polyesters produced in large amounts by different microorganisms, but not by E.coli. Here we expressed the bacterial polyhydroxybutarate (PHB) pathway of Halomonas boliviensis in E. coli using phbA and phbB genes to produce the monomer 3-HB. Acetyl coenzyme A (AcCoA) is the intermediate of the carbon metabolism and precursor of the PHB pathway. The objective is to design a process to produce 3-HB in E.coli with high productivity which is a first step to further produce different quality polyesters. Production of the wild type microorganisms takes place under nutrient deficient conditions and excess of a carbon source. The hypothesis is that directing the flux to the desired product (3-HB) by cultivations with controlled feed of nitrogen or phosphorous will increase the flux to the precursor (AcCoA) and minimize the formation of byproducts This will be done by: Fed-Batch cultivations with an excess of glucose and nitrogen limitation. Excess glucose will give a surplus of NADH that will inhibit citrate synthase and the AcCoA produced will instead be used for 3-HB production Fed-Batch cultivations with an excess of glucose and phosphorous limitation. Excess glucose will give a surplus of NADH that will inhibit citrate synthase and the AcCoA produced will instead be used for 3-HB production. Also the limitation of phosphorous activates the methylglyoxal pathway also leading to the more energy favorable production of AcCoA than central glycolisis. http://dx.doi.org/10.1016/j.nbt.2014.05.1985