• Microbiology and Biotechnology Letters
• /
• v.18 no.1
• /
• pp.103-108
• /
• 1990

The enzymatic studies on the methylglyoxal metabolism in yeast and bacterial cells indicated that organisms are equipped with the common and manifold systems for the detoxification of methylglyoxal. Among these systems, the glyoxalase I is the most important route for methylglyoxal detoxification. The molecular structure of glyoxalase I is apparently distinct from the enzyme sources, and zinc ion is an essential cofactor in enzyme activity. The gene for Pseudomonas putida glyoxalase I functioned as a scavenger of methylglyoxal and regulated the cell size of the bacterium. Comparison of the nucleotide sequence of the P. putida glyoxalase I gene with the N-terminal amino acid sequence of the purified enzyme revealed that the N-terminal methionine residue was removed after translation. Possible physiological role of glyoxalase I was also discussed.

• Proceedings of the Korea Crystallographic Association Conference
• /
• 2003.05a
• /
• pp.17-17
• /
• 2003

tRNA (m¹ G37) methyltransferase (TrmD) catalyze s the trans for of a methyl group from S-adenosyl-L-methionine (AdoMet) to G/sup 37/ within a subset of bacterial tRNA species, which have a residue G at 36th position. The modified guanosine is adjacent to and 3' of the anticodon and is essential for the maintenance of the correct reading frame during translation. We have determined the first crystal structure of TrmD from Haemophilus influenzae, as a binary complex with either AdoMet or S-adenosyl-L-homocysteine (AdoHcy), as a ternary complex with AdoHcy/phosphate, and as an apo form. The structure indicates that TrmD functions as a dimer (Figure 1). It also suggests the binding mode of G/sup 36/G/sup 37/ in the active site of TrmD and catalytic mechanism. The N-terminal domain has a trefoil knot, in which AdoMet or AdoHcy is bound in a novel, bent conformation. The C-terminal domain shows a structural similarity to DNA binding domain of trp or tot repressor. We propose a plausible model for the TrmD₂-tRNA₂ complex, which provides insights into recognition of the general tRNA structure by TrmD (Figure 2).

• Journal of Microbiology and Biotechnology
• /
• v.23 no.3
• /
• pp.329-334
• /
• 2013

Uridinediphospho-N-acetylglucosamine enolpyruvyl transferase (MurA, E.C. 2.5.1.7) is an essential bacterial enzyme that catalyzes the first step of the cell wall biosynthetic pathway, which involves the transfer of an enolpyruvyl group from phosphoenolpyruvate to uridinediphospho-Nacetylglucosamine. In this study, novel inhibitors of Haemophilus influenzae MurA (Hi MurA) were identified using high-throughput screening of a chemical library from the Korea Chemical Bank. The identified compounds contain a quinoline moiety and have much lower effective inhibitory concentrations ($IC_{50}$) than fosfomycin, a wellknown inhibitor of MurA. These inhibitors appear to covalently modify the sulfhydryl group of the active site cysteine (C117), since the C117D mutant Hi MurA was not inhibited by these compounds and excess dithiothreitol abolished their inhibitory activities. The increased mass value of Hi MurA after treatment with the identified inhibitor further confirmed that the active-site cysteine residue of Hi MurA is covalently modified by the inhibitor.

• Bulletin of the Korean Chemical Society
• /
• v.24 no.12
• /
• pp.1799-1804
• /
• 2003

Acetolatate synthase(ALS) catalyzes the first common step in the biosynthesis of valine, leucine, isoleucine in plants and microorganisms. ALS is the target of several classes of herbicides, including the sulfonylureas, the imidazolinones, and the triazolopyrimidines. To elucidate the roles of arginine residues in tobacco ALS, chemical modification and site-directed mutagenesis were performed. Recombinant tobacco ALS was expressed in E. coli and purified to homogeneity. The ALS was inactivated by arginine specific reagents, phenylglyoxal and 2,3-butanedione. The rate of inactivation was a function of the concentration of modifier. The inactivation by butanedione was enhanced by borate, and the inactivation was reversible on removal of excess butanedione and borate. The substrate pyruvate and competitive inhibitors fluoropyruvate and phenylpyruvate protected the enzyme against inactivation by both modifiers. The mutation of well-conserved Arg198 of the ALS by Gln abolished the enzymatic activity as well as the binding affinity for cofactor FAD. However, the mutation of R198K did not affect significantly the binding of FAD to the enzyme. Taken together, the results imply that Arg198 is essential for the catalytic activity of the ALS and involved in the binding of FAD, and that the positive charge of the Arg is crucial for the interaction with negatively charged FAD.

• Food Science and Biotechnology
• /
• v.16 no.1
• /
• pp.37-42
• /
• 2007

The biotransformation of monoterpenic agro-industrial wastes (turpentine oil and essential orange oil) was studied. More than 40 fungal strains were isolated from Brazilian tropical fruits and eucalyptus trees and screened for biotransformation of the waste substrates. Solid phase microextraction was used to monitor the presence of volatile compounds in the headspaces of sporulated surface cultures. The selected strains were submitted to submerged liquid culture. The biotransformation of R-(+)-limonene and ${\alpha},\;{\beta}-$ pinenes from the oils resulted in ${\alpha}-terpineol$ and perillyl alcohol, and verbenol and verbenone, respectively, as the main products. The selected strains were also placed in contact with ${\alpha}-$ and ${\beta}-$ pinenes standards. It was confirmed that verbenol, verbenone, and ${\alpha}-terpineol$ were biotransformation products from the terpenes. A concentration of 90 mg/L of verbenone was achieved by Penicillium sp. 2360 after 3 days of biotransformation.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
• /
• 2003.05c
• /
• pp.149-153
• /
• 2003

Chemical mechanical polishing (CMP) technology for global planarization of multi-level inter-connection structure has been widely studied for the next generation devices. CMP process has been paid attention to planarized pre-metal dielectric (PMD), inter-layer dielectric (ILD) interconnections. Expecially, shallow trench isolation (STI) used to CMP process on essential. Recently, the direct STI-CMP process without the conventional complex reverse moat etch process has established by using slurry additive with the high selectivity between $SiO_2$ and $Si_3N_4$ films for the purpose of process simplification and n-situ end point detection(EPD). However, STI-CMP process has various defects such as nitride residue, tom oxide and damage of silicon active region. To solve these problems, in this paper, we studied the planarization characteristics using a high selectivity slurry(HSS). As our experimental results, it was possible to achieve a global planarization and STI-CMP process could be dramatically simplified. Also we estimated the reliability through the repeated tests with the optimized process conditions in order to identify the reproducibility of HSS STI-CMP process.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.5
• /
• pp.838-844
• /
• 2001

The enzyme Fuc aldolase from Methanococcus jannaschii that catalyzes the aldol condensation of DHAP and L-lactaldehyde to give fuculose-1-phosphate was inactivated by DEP. The inactivation was pseudo first-order in the enzyme and DEP, which was biphasic. A pseudo second-order rate constant of 120$M^{-1}min^{-1}$ was obtained at pH 6.0 and $25{\circ}C$. Quantifying the increase in absorbance at 240nm showed that four histidine residues per subunit were modified during the nearly complete inactivation. The statistical analysis and the time course of the modification suggested that two or three histidine residues were essential for activity. The rate of inactivation was dependent on the pH, and the pH inactivation data implied the involvement of the amino acid residue with a $pK_a$ value of 5.7. Fuc aldolase was protected against DEP inactivation by DHAP, indicating that the histidine residues were located at the active site of Fuc aldolase. DL-Glyceraldehyde, as an alternative substrate to L-lactaldehyde, showed no specific protection for the Fuc aldolase.

• Journal of the Korean Dietetic Association
• /
• v.25 no.1
• /
• pp.44-58
• /
• 2019

Predicting the number of meals in a foodservice organization is an important decision-making process that is essential for successful food production, such as reducing the amount of residue, preventing menu quality deterioration, and preventing rising costs. Compared to other demand forecasts, the menu of dietary personnel includes diverse menus, and various dietary supplements include a range of side dishes. In addition to the menus, diverse subjects for prediction are very difficult problems. Therefore, the purpose of this study was to establish a method for predicting the number of meals including predictive modeling and considering various factors in addition to menus which are actually used in the field. For this purpose, 63 variables in eight categories such as the daily available number of people for the meals, the number of people in the time series, daily menu details, weekdays or seasons, days before or after holidays, weather and temperature, holidays or year-end, and events were identified as decision variables. An ensemble model using six prediction models was then constructed to predict the number of meals. As a result, the prediction error rate was reduced from 10%~11% to approximately 6~7%, which was expected to reduce the residual amount by approximately 40%.

• Korean Journal of Environmental Agriculture
• /
• v.37 no.4
• /
• pp.283-290
• /
• 2018

BACKGROUND: Pesticide residue analysis is an essential activity in order to establish the food safety of agricultural products. Analytical approaches to the food safety are required to meet internationally the guideline of Codex (Codex Alimentarius Commission, CAC/GL 40). In this study, we developed a liquid chromatograph-tandem mass spectrometer (LC-MS/MS) method to determine the herbicide clopyralid in food matrixes. METHODS AND RESULTS: Clopyralid was extracted with aqueous acetonitrile containing formic acid and the extracts were mixed in a citrate buffer consisted of magnesium sulfate anhydrous, NaCl, sodium citrate dihydrate and disodium hydrogencitrate sesquihydrate followed by centrifugation. The supernatants were filtered through a nylon membrane filter and used for the analysis of clopyralid. The method was validated by accuracy and precision experiments on the samples fortified at 3 different levels of clopyralid. LC-MS/MS in positive mode was employed to quantitatively determine clopyralid in the food samples. Matrix-matched calibration curves were inearranged from 0.001 to 0.25 mg/kg with r2 > 0.994. The limits of detection and quantification were determined to be 0.001 and 0.01 mg/kg, respectively. There covery values of clopyralid for tified at 0.01 mg/kg in the control samples ranged from approximately 82 to 106% with relative standard deviations below 2 0%. CONCLUSION: The method developed in this study meets successfully the Codex guideline for pesticide residue analysis in food samples. This, the method could be applicable to determine pesticides in foods produced domestically and internationally.

• Journal of Life Science
• /
• v.17 no.9 s.89
• /
• pp.1182-1190
• /
• 2007

The gene encoding for isoamylase of the Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 was cloned and expressed into Escherichia coli $DH5{\alpha}$. Isoamylase catalyzes the hydrolysis of ${\alpha}-1,6-glycosidic$ linkages specifically in amylopectin, glycogen, and derived oligosaccharides, while the enzyme did not hydrolyze ${\alpha}-1,4-glycosidic$ linkages of amylose. The isoamylase gene (glgX) had an open reading frame of 1,977 bp encoding 658 amino acid residues with a calculated molecular weight of 74,188 Da. The molecular weight of the enzyme was also estimated to be 74 kDa by activity staining of a SDS-PA gel. The mature GlgX had a calculated pI of 4.91. Isoamylase from Pcc LY34 had 70% amino acid identity with isoamylase from Pectobacterium chrysanthemi and contained the four regions conserved among all amylolytic enzymes. The isoamylase was optimally active at pH 7.0 and $40^{\circ}C$. GlgX was $Ca^{2+}-dependent$. The changes of Asp-335, Glu-370, and Asp-442 into Ala, respectively, using site-directed mutagenesis techniques showed that three residues are essential to isolamyalse (GlgX) activity. The sequences around those residues were highly conserved in isoamylase of different origins and GlgX of the glg operon in glycongen biosynthesis.