Seo, Gyeong-Hee;Yun, Jung-Hyun;Chung, Sun-Kyung;Park, Woo-Po;Lee, Dong-Sun
Food Science and Biotechnology
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v.15
no.2
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pp.168-172
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2006
Soy sauce was fermented at $20^{\circ}C$ for 100 days in onggi containers (ethnic Korean earthenware) which had been fabricated using three different glazing treatments: unglazed, glazed only on the outside, and glazed on both surfaces. The changes in microstructure and permeability characteristics of onggi containers were examined after fermentation of soy sauce. The effect of repeated use of onggi containers on the fermentation was analyzed by the contact between an aqueous model solution and the onggi containers used once for soy sauce fermentation. The levels of reducing sugar and free amino acids produced from the dissolved starch and protein, respectively, in the solution were compared between the new and reused onggi containers. The moisture permeance and gas permeabilities of the onggi jars were progressively reduced with continuing use for soy sauce fermentation, probably due to clogging of micropores by solid materials. After having been used once for fermentation, the microbial cells and/or enzymes immobilized on the surface or in the micropores of the onggi containers seemed to contribute to accelerating the hydrolytic reactions of starch and protein.
The recent progress on metabolic systems engineering was reviewed based on our recent research results in terms of (1) metabolic signal flow diagram approach, (2) metabolic flux analysis (MFA) in particular with intracellular isotopomer distribution using NMR and/or GC-MS, (3) synthesis and optimization of metabolic flux distribution (MFD), (4) modification of MFD by gene manipulation and by controlling culture environment, (5) metabolic control analysis (MCA), (6) design of metabolic regulation structure, and (7) identification of unknown pathways with isotope tracing by NMR. The main characteristics of metabolic engineering is to treat metabolism as a network or entirety instead of individual reactions. The applications were made for poly-3-hydroxybutyrate (PHB) production using Ralstonia eutropha and recombinant Escherichia coli, lactate production by recombinant Saccharomyces cerevisiae, pyruvate production by vitamin auxotrophic yeast Toluropsis glabrata, lysine production using Corynebacterium glutamicum, and energetic analysis of photosynthesic microorganisms such as Cyanobateria. The characteristics of each approach were reviewed with their applications. The approach based on isotope labeling experiments gives reliable and quantitative results for metabolic flux analysis. It should be recognized that the next stage should be toward the investigation of metabolic flux analysis with gene and protein expressions to uncover the metabolic regulation in relation to genetic modification and/ or the change in the culture condition.
Proceedings of the Korean Society for Applied Microbiology Conference
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2004.06a
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pp.60-61
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2004
Metabolic engineering is now a well established discipline, used extensively to determine and execute rational strategies of strain development to improve the performance of micro-organisms employed in industrial fermentations. The basic principle of this approach is that performance of the microbial catalyst should be adequately characterised metabolically so as to clearlyidentify the metabolic network constraints, thereby identifying the most probable targets for genetic engineering and the extent to which improvements can be realistically achieved. In order to harness correctly this potential, it is clear that the physiological analysis of each strain studied needs to be undertaken under conditions as close as possible to the physico-chemical environment in which the strain evolves within the full-scale process. Furthermore, this analysis needs to be undertaken throughoutthe entire fermentation so as to take into account the changing environment in an essentially dynamic situation in which metabolic stress is accentuated by the microbial activity itself, leading to increasingly important stress response at a metabolic level. All too often these industrial fermentation constraints are overlooked, leading to identification of targets whose validity within the industrial context is at best limited. Thus the conceptual error is linked to experimental design rather than inadequate methodology. New tools are becoming available which open up new possibilities in metabolic engineering and the characterisation of complex metabolic networks. Traditionally metabolic analysis was targeted towards pre-identified genes and their corresponding enzymatic activities within pre-selected metabolic pathways. Those pathways not included at the onset were intrinsically removed from the network giving a fundamentally localised vision of pathway functionality. New tools from genome research extend this reductive approach so as to include the global characteristics of a given biological model which can now be seen as an integrated functional unit rather than a specific sub-group of biochemical reactions, thereby facilitating the resolution of complexnetworks whose exact composition cannot be estimated at the onset. This global overview of whole cell physiology enables new targets to be identified which would classically not have been suspected previously. Of course, as with all powerful analytical tools, post-genomic technology must be used carefully so as to avoid expensive errors. This is not always the case and the data obtained need to be examined carefully to avoid embarking on the study of artefacts due to poor understanding of cell biology. These basic developments and the underlying concepts will be illustrated with examples from the author's laboratory concerning the industrial production of commodity chemicals using a number of industrially important bacteria. The different levels of possibleinvestigation and the extent to which the data can be extrapolated will be highlighted together with the extent to which realistic yield targets can be attained. Genetic engineering strategies and the performance of the resulting strains will be examined within the context of the prevailing experimental conditions encountered in the industrial fermentor. Examples used will include the production of amino acids, vitamins and polysaccharides. In each case metabolic constraints can be identified and the extent to which performance can be enhanced predicted
Statement of the problem: Implant systems result in gaps and cavities between implant and abutment that can act as a trap for bacteria and thus possibly cause inflammatory reactions in the peri-implant soft tissues. Purpose: Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Treponema denticola, and Aggregatibacter actinomycetemcomitans, related to implant-abutment interface microleakage. Material and methods: Samples were taken from 27 subjects with sterilized paper points and were transported in $1{\times}PBS$. The detection of periodontopathogens were performed by polymerase chain reaction with species-specific primers based on 16S rDNA. Results: Our data showed that the detection rate of P. gingivalis and P. intermedia in implant fixture was 59% and 82% in patients respectively. Detection rate of P. gingivalis and P. intermedia in implant crevice was 44% and 82% in patients. Detection rate of P. gingivalis and P. intermedias in tongue was 82% and 82% in patients. Conclusion: Current implant systems cannot safely prevent microbial leakage and bacterial colonization of the inner part of the implant.
Water distribution system (WDS) is exposed to various water quality incidents during its operation. This study utilized Quantitative Microbial Risk Assessment (QMRA) to analyze the risk associated with potential virus intrusion in WDSs. Additionally, the study determined the location and operation of rechlorination facilities to minimize potential risk. In addition, water quality resilience was calculated to confirm that the chlorine concentration maintains within the target range (0.1-1.0 mg/L) during normal operation. Hydraulic analysis was performed using EPANET, while EPANET-MSX was linked to simulate the reactions between viruses and chlorine. The proposed methodology was applied to the Bellingham network in the United States, where rechlorination facilities capable of injecting chlorine concentrations ranging from 0.5 mg/L to 1.0 mg/L were considered. Results indicated that without rechlorination facilities, the Average risk was 0.0154. However, installing rechlorination facilities and injecting chlorine at a concentration of 1.0 mg/L could reduce the Average risk to 39.1%. It was observed that excessive chlorine injection through rechlorination facilities reduced water quality resilience. Consequently, a rechlorination facility with a concentration of 0.5 mg/L was selected, resulting in a reduction of approximately 20% in average risk. This study provides insights for designing rechlorination facilities to enhance preparedness against potential virus ingress in the future.
Proceedings of the Korean Society of Life Science Conference
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2001.11a
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pp.3-8
/
2001
Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids, IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5(, XL1-Blue, and JM101) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter (PBAD) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters (Ptrc and Plac, respectively) on medium-copy and high-copy plasmids, Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 mM, cells expressing both dxs and dxr from PBAD on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plamid revealed that lycopene production was highest in XL1-Blue.
Cytochrome P4502C19 (CYP2C19) is one of human polymorphic xenobiotic-metabolizing enzymes. The enzyme has been reported to catalyze more than 70 substrates, involving more than 100 reactions. These include several classes of therapeutic agents (e.g. anti-microbial. cardiovascular, psycho-active, etc.), sex hormones and insecticides. Associations of the CYP2C19 genotype/phenotype with individual differences in drug efficacy (e.g. diazepam, omeprazole, proguanil) and toxicity (e.g. mephenytoin, barbiturates) have been documented by many investigators. At least 11 allelic variants of CYP2C19 gene were reported to date. Most of the mutant alleles found in the poor metabolizer (PM) led to the production of truncated and/or inactive proteins. Except for the exon 6, single-nucleotide mutations were reported in all nine exons of the gene. Genetic polymorphism of CYP2C19 shows marked interethnic variation with the population frequencies of PM phenotype ranging from 1∼2% up to more than 50%. The prevalence of CYP2C19 PM tends to be higher in Asian and certain Pacific Islanders than other race or ethnic specificity. Genotyping results of CYP2C19 also revealed that there are different proportions of individual mutant alleles among ethnic populations. This may, in part, explains the interethnic difference in the metabolism of certain drugs (i.e. diazepam), though they were from the same CYP2C19 phenotype. Recently, our research group has studied the genotype and phenotype of CYP2C19 and found that the PM frequency (7∼8%) in Thais is lower than other Asian populations. Molecular and clinical impacts of this finding warrant to further investigation.
The enzymes were immobilized by treating the microbial cells in 0.05% chitosan and 0.28% glutaraldehyde solution. The activity of immobilized cell was about 535 IGIC/g. Glucose isomerase was purified by 6.5 times after homogenization using 60% $(NH_4)_2S0_4$ fractionation, DEAE-cellulose and Sephadex G-150 gel filtration. The molecular weight of enzyme was about 140,000 when it was measured by HPLC and the purified enzyme had only one band by electrophoresis. It showed good enzyme activity at pH 7.5 and $75^{\circ}C$. The optimum conditions for enzyme reactions were shifted to pH 7.0 and $80^{\circ}C$ when the enzyme was immobilized. The enzyme reaction was activated by the addition of 5~10 mM magnesium ion and the thermostability was improved by the addition of 0.25 mM cobalt ion. The enzyme activity was competitively inhibited by sugar alcohols.
A small amount of milk is sold as 'untreated' or raw in the US; the two most commonly used heat-treatments for milk sold in retail markets are pasteurization (LTLT, low-temperature long time; HTST, high-temperature short time) and sterilization (UHT, ultra-high temperature). These treatments extend the shelf life of milk. The main purpose of heat treatment is to reduce pathogenic and perishable microbial populations, inactivate enzymes, and minimize chemical reactions and physical changes. Milk UHT processing combined with aseptic packaging has been introduced to produce shelf-stable products with less chemical damage than sterile milk in containers. Two basic principles of UHT treatment distinguish this method from in-container sterilization. First, for the same germicidal effect, HTST treatments (as in UHT) use less chemicals than cold-long treatment (as in in-container sterilization). This is because Q10, the relative change in the reaction rate with a temperature change of $10^{\circ}C$, is lower than the chemical change during bacterial killing. Based on Q10 values of 3 and 10, the chemical change at $145^{\circ}C$ for the same germicidal effect is only 2.7% at $115^{\circ}C$. The second principle is that the need to inactivate thermophilic bacterial spores (Bacillus cereus and Clostridium perfringens, etc.) determines the minimum time and temperature, while determining the maximum time and temperature at which undesirable chemical changes such as undesirable flavors, color changes, and vitamin breakdown should be minimized.
Kim, Ji Yeon;Kim, Kwang Joon;Park, Sang-Wook;Bang, Joon Seok;Lee, Wonjae
Korean Journal of Clinical Pharmacy
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v.24
no.3
/
pp.219-228
/
2014
Objectives: This paper was aimed to investigate the adhesion control standards of pain relieving patch (PRP) drugs and to survey it's adverse effects on the skin of patients for safe use of PRP drugs. Methods: In this study, the related documents of PRP drugs of Korea pharmacopoeia (KP), United States pharmacopoeia (USP), Japanese Pharmacopoeia (JP), European pharmacopoeia (EP), and information web sites of the Ministry of Food and Drug Safety (MFDS) were surveyed. Also, the past and current labeling of PRP drugs marketed in the pharmacy was investigated and compared. Results: In KP and JP, the lower limit standard for PRP's adhesion control is established, but the upper limit standard is not designated. In USP and EP, neither the lower nor upper limit standard is established. The main reasons of skin adverse effects are considered as inherent adverse reactions of the applied drugs for PRP. Another reason is involved in patient's medication mistakes related to PRP's adhesion control, respiratory depression of skin according to physical skin closure, and microbial growth, etc. Conclusion: For safe use of PRP drugs, we proposed ensured guidelines like additional instructions of pharmacist's prescription and detailed labeling systems for usage of PRP drugs applied on skin.
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