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Comparative proteomic analysis of four biotechnologically important Escherichia coli strains for rational host strain selection
S48
Abstracts / Journal of Biotechnology 136S (2008) S22–S71
I1-P-069 Inhibitory efficacy of Lactococcus lactis on food born pathogens and food spoilage bacteria Saeed Mirdamadi 1,2,∗ , Shadi Agha Ghazvini 3 , Mohamad Reza Ehsani 4 , Masoud Falahpour 5 , Say-yed Hesameddin Tafreshi 6 1
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran Biotechnology Department of Iranian Research Organization for Science and Technology (IROST), Tehran, Iran 3 Azad University, Science and Research Branch, Pasteur Institute of Iran, Tehran, Iran 4 Tehran University, Food Science Department, Pasteur Institute of Iran, Tehran, Iran 5 Iranian Research Organization for Science & Technology, Pasteur Institute of Iran, Tehran, Iran 6 Injection Solutions Department, Research and Production Complex, Pasteur Institute of Iran, Tehran, Iran 2
E-mail address: Mirdamadi [email protected] (S. Mirdamadi). The growth inhibitory effects of Lactococcus lactis subsp. lactis ATCC 11454 against Listeria monocytogenes, Staphylococcus aureus and Eschericha coli was investigated during associated culture. The growth rate of Listeria monocytogenes, S. aureus and E. coli was studied in the presence and absence of Lactococcus lactis for 48 h at 30 ◦ C (Bielecka et al., 1998; Millette et al., 2006). The Lactococcus lactis inhibited growth of Listeria monocytogenes and S. aureus significantly. The growth rate of E. coli was remained steady by Lactococcus lactis in the media. In our research, the maximum production of nisin was observed in lag phase of pathogenic bacteria growth (468 IU mL−1 ) (Ogawa et al., 2001). On the other hand, pH of cultures decreased from 6.5 to 4.70 during associated culture. Since nisin does not show any inhibitory impacts on E. coli, so reduction in pH may be the cause of inhibitory against E. coli. Listeria was the most sensitive bacteria in this study. The kinetic growth rate of Listeria and Staphylococcus was investigated (Millette et al., 2006). So, kinetics of growth and inhibitory effects of Lactococcus lactis against pathogenic and spoilage food bacteria were studied in this paper. References Bielecka, M., Biedrzycka, E., Biedrzycka, E., Smoragiewicz, W., Smieszek, M., 1998. Interaction of Bifidobacterium and Salmonella during associated growth. Int. J. Food Microbiol. 45, 151–155. Millette, M., Luquet, F.M., Lacroix, 2006. In vitro growth control of selected pathogens by Lactobacillus acidophilus- and Lactobacillus casei-fermented milk. Lett. Appl. Microbiol. 44, 314–319. Ogawa, M., Shimizu, K., Nomoto, K., Tanaka, R., Hamabata, T., Yamasaki, S., Takeda, T., Takeda, Y., 2001. Inhibition of in vitro growth of shiga toxin-producing Escherichia coli O157:H7 by probiotic Lactobacillus strains due to production of lactic acid. Int. J. Food Microbiol. 68, 135–140.
doi:10.1016/j.jbiotec.2008.07.098 I1-P-070 In vitro evaluation controlled release study for Metformin hydrochloride polymeric hydrogel matrixes
in several ratios (25%, 34%, 50%, w/w).The two polymeric matrix systems were prepared by compression molding and dip coating process, respectively. The release rate of Metformin hydrochloride from polymeric matrix devices was studied by using UV technique at constant temperature (37 ◦ C). The effect of pH on the release rate of Metformin hydrochloride in (PBS, pH 7.4), SGF, and SIF was studied. The swelling behavior of some prepared samples was also studied, the result showed greater increase in swelling ratio at lower pH values. doi:10.1016/j.jbiotec.2008.07.099 I1-P-074 Comparative proteomic analysis of four biotechnologically important Escherichia coli strains for rational host strain selection Xiao-Xia Xia 1 , Sang Yup Lee 1,2,∗ 1
Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program) and BioProcess Engineering Research Center, Center for Systems and Synthetic Biotechnology, and Institute for the BioCentury, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea 2 Department of Bio and Brain Engineering, and Bioinformatics Research Center, KAIST, Daejeon, Republic of Korea E-mail address: [email protected] (S.Y. Lee). Escherichia coli B and K12 derivatives are commonly used in laboratory and bioindustry due to their fast growing and easy cultivation. After sequencing the complete genome of parent B and K12 strains, we have the demanding task of investigating the proteomes of these strains and obtaining valuable physiological information through comparative proteomics (Blattner and Plunkett, 1997; Xia et al., 2008). Here, using two-dimensional gel electrophoresis coupled with MS/MS, we profiled the proteomes of an E. coli B derivative and three K12 derivatives grown with a defined medium at exponential phase and stationary phase, respectively. Over 100 proteins were differentially expressed, including those involved in cellular processes and stress responses, and those in the main metabolic pathways (glycolysis, TCA cycle, gluconeogenesis and fermentative pathway) and the synthesis pathways (amino acid biosynthesis, fatty acid biosynthesis, nucleotide synthesis and energy synthesis). A correlation between the proteome profiling and metabolic pathway analysis revealed significant metabolic differences among the four strains, which provided valuable information for the production of various primary metabolites. Also, analysis of amino acid synthesis and protein processing shed light on rational host selection for recombinant protein production. In conclusion, comparative proteomic analysis revealed systematic metabolic diversity among the four strains studied, and provided valuable information for rational host strain.
Emad M. Abdelrazaq
References
Department of Chemistry, College of Science, University of Basrah, Basrah, Iraq
Blattner, F.R., Plunkett III, G., 1997. The complete genome sequence of Escherichia coli K-12. Science 277, 1453–1462. Xia, X.-X., Han, M.-J., Lee, S.Y., Yoo, J.-S., 2008. Comparison of the extracellular proteomes of Escherichia coli B and K-12 strains during high cell density cultivation. Proteomics 8 (1), 2089–2103.
The hydroxyethylcellulose hydrogels were prepared by its chemical crosslinking using glyceraldehyde, and tetramethylolurea resin in the presence of sulfuric acid. Several tablets were prepared as polymeric matrix and polymeric reservoir systems from hydroxyethylcellulose hydrogels, loaded with Metformin hydrochloride
doi:10.1016/j.jbiotec.2008.07.100
Abstracts / Journal of Biotechnology 136S (2008) S22–S71
I1-P-069 Inhibitory efficacy of Lactococcus lactis on food born pathogens and food spoilage bacteria Saeed Mirdamadi 1,2,∗ , Shadi Agha Ghazvini 3 , Mohamad Reza Ehsani 4 , Masoud Falahpour 5 , Say-yed Hesameddin Tafreshi 6 1
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran Biotechnology Department of Iranian Research Organization for Science and Technology (IROST), Tehran, Iran 3 Azad University, Science and Research Branch, Pasteur Institute of Iran, Tehran, Iran 4 Tehran University, Food Science Department, Pasteur Institute of Iran, Tehran, Iran 5 Iranian Research Organization for Science & Technology, Pasteur Institute of Iran, Tehran, Iran 6 Injection Solutions Department, Research and Production Complex, Pasteur Institute of Iran, Tehran, Iran 2
E-mail address: Mirdamadi [email protected] (S. Mirdamadi). The growth inhibitory effects of Lactococcus lactis subsp. lactis ATCC 11454 against Listeria monocytogenes, Staphylococcus aureus and Eschericha coli was investigated during associated culture. The growth rate of Listeria monocytogenes, S. aureus and E. coli was studied in the presence and absence of Lactococcus lactis for 48 h at 30 ◦ C (Bielecka et al., 1998; Millette et al., 2006). The Lactococcus lactis inhibited growth of Listeria monocytogenes and S. aureus significantly. The growth rate of E. coli was remained steady by Lactococcus lactis in the media. In our research, the maximum production of nisin was observed in lag phase of pathogenic bacteria growth (468 IU mL−1 ) (Ogawa et al., 2001). On the other hand, pH of cultures decreased from 6.5 to 4.70 during associated culture. Since nisin does not show any inhibitory impacts on E. coli, so reduction in pH may be the cause of inhibitory against E. coli. Listeria was the most sensitive bacteria in this study. The kinetic growth rate of Listeria and Staphylococcus was investigated (Millette et al., 2006). So, kinetics of growth and inhibitory effects of Lactococcus lactis against pathogenic and spoilage food bacteria were studied in this paper. References Bielecka, M., Biedrzycka, E., Biedrzycka, E., Smoragiewicz, W., Smieszek, M., 1998. Interaction of Bifidobacterium and Salmonella during associated growth. Int. J. Food Microbiol. 45, 151–155. Millette, M., Luquet, F.M., Lacroix, 2006. In vitro growth control of selected pathogens by Lactobacillus acidophilus- and Lactobacillus casei-fermented milk. Lett. Appl. Microbiol. 44, 314–319. Ogawa, M., Shimizu, K., Nomoto, K., Tanaka, R., Hamabata, T., Yamasaki, S., Takeda, T., Takeda, Y., 2001. Inhibition of in vitro growth of shiga toxin-producing Escherichia coli O157:H7 by probiotic Lactobacillus strains due to production of lactic acid. Int. J. Food Microbiol. 68, 135–140.
doi:10.1016/j.jbiotec.2008.07.098 I1-P-070 In vitro evaluation controlled release study for Metformin hydrochloride polymeric hydrogel matrixes
in several ratios (25%, 34%, 50%, w/w).The two polymeric matrix systems were prepared by compression molding and dip coating process, respectively. The release rate of Metformin hydrochloride from polymeric matrix devices was studied by using UV technique at constant temperature (37 ◦ C). The effect of pH on the release rate of Metformin hydrochloride in (PBS, pH 7.4), SGF, and SIF was studied. The swelling behavior of some prepared samples was also studied, the result showed greater increase in swelling ratio at lower pH values. doi:10.1016/j.jbiotec.2008.07.099 I1-P-074 Comparative proteomic analysis of four biotechnologically important Escherichia coli strains for rational host strain selection Xiao-Xia Xia 1 , Sang Yup Lee 1,2,∗ 1
Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program) and BioProcess Engineering Research Center, Center for Systems and Synthetic Biotechnology, and Institute for the BioCentury, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea 2 Department of Bio and Brain Engineering, and Bioinformatics Research Center, KAIST, Daejeon, Republic of Korea E-mail address: [email protected] (S.Y. Lee). Escherichia coli B and K12 derivatives are commonly used in laboratory and bioindustry due to their fast growing and easy cultivation. After sequencing the complete genome of parent B and K12 strains, we have the demanding task of investigating the proteomes of these strains and obtaining valuable physiological information through comparative proteomics (Blattner and Plunkett, 1997; Xia et al., 2008). Here, using two-dimensional gel electrophoresis coupled with MS/MS, we profiled the proteomes of an E. coli B derivative and three K12 derivatives grown with a defined medium at exponential phase and stationary phase, respectively. Over 100 proteins were differentially expressed, including those involved in cellular processes and stress responses, and those in the main metabolic pathways (glycolysis, TCA cycle, gluconeogenesis and fermentative pathway) and the synthesis pathways (amino acid biosynthesis, fatty acid biosynthesis, nucleotide synthesis and energy synthesis). A correlation between the proteome profiling and metabolic pathway analysis revealed significant metabolic differences among the four strains, which provided valuable information for the production of various primary metabolites. Also, analysis of amino acid synthesis and protein processing shed light on rational host selection for recombinant protein production. In conclusion, comparative proteomic analysis revealed systematic metabolic diversity among the four strains studied, and provided valuable information for rational host strain.
Emad M. Abdelrazaq
References
Department of Chemistry, College of Science, University of Basrah, Basrah, Iraq
Blattner, F.R., Plunkett III, G., 1997. The complete genome sequence of Escherichia coli K-12. Science 277, 1453–1462. Xia, X.-X., Han, M.-J., Lee, S.Y., Yoo, J.-S., 2008. Comparison of the extracellular proteomes of Escherichia coli B and K-12 strains during high cell density cultivation. Proteomics 8 (1), 2089–2103.
The hydroxyethylcellulose hydrogels were prepared by its chemical crosslinking using glyceraldehyde, and tetramethylolurea resin in the presence of sulfuric acid. Several tablets were prepared as polymeric matrix and polymeric reservoir systems from hydroxyethylcellulose hydrogels, loaded with Metformin hydrochloride
doi:10.1016/j.jbiotec.2008.07.100