• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.494-497
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• 1998

Deodorization of soybean oil was carried out at a temperature range of $220{\sim}280^{\circ}C$ for 1 or 2hrs. HPSEC(high performance size exclusion chromatography) method was used for the analysis of dimeric acids in deodorized soybean oil. Dimeric acid moieties were produced through the deodorization at $240{\sim}280^{\circ}C$ for 2 hrs. The amount of dimeric acids produced was found to be in the range of 0.36 to 3.39%. Dimeric acids were not detected under the deodorizing condition of $240^{\circ}C,\;2\;hrs\;or\;250^{\circ}C,\;1\;hr$. The soybean oil deodorized in these conditions had good quality by the color and viscosity tests. The best condition of minimizing the formation of dimeric acids in soybean oils was the deodorizing conditions of $240^{\circ}C,\;2\;hrs\;or\;250^{\circ}C,\;1hr$.

• BMB Reports
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• v.32 no.2
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• pp.181-188
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• 1999

4-Aminobutyrate aminotransferase plays an essential role in the 4-aminobutyric acid shunt, converting 4-aminobutyrate to succinic semialdehyde. Recombinant 4-aminobutyrate aminotransferases were overexpressed as their catalytically active forms in E. coli by coproduction with thioredoxin and their solubilities were also dramatically increased. In order to study the structural and functional aspects of the C-terminal domain of brain 4-aminobutyrate aminotransferase, we have constructed a C-terminal mutant of pig brain 4-aminobutyrate aminotransferase and analyzed the functional and structural roles of C-terminal amino acids residues on the enzyme. The deletion of five amino-acid residues from C-terminus did not interfere with the kinetic parameters and functional properties of the enzyme. Also, the deletion did not affect the dimeric structure of the protein aligned along the subunit interface at neutral pH. However, the deletion of the C-terminal region of the protein changed the stability of its dimeric structure at acidic pH. The dissociation of the enzyme acidic, facilitated by the deletion of five amino acids from C-terminus, abolished the catalytic activity.

• Proceedings of the PSK Conference
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• 2002.04a
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• pp.186.2-186.2
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• 2002

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.132-137
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• 1997

A novel protein (HEAAE, High Essential Amino Acid Encoding Protein), rich in essential amino acids ($75{\%}$ of total), was designed and constructed in our laboratory. The designed peptides were analyzed by SYBLE and stable secondary and tertiary structures were predicted. The monomeric form (HEAAE-1) of the protein consists of 20 amino acid residues with four additional amino acids comprising a potential ${\beta}$-turn (HEAAE-4). Size exclusion analysis demonstrated that the monomer is self-aggregates in aqueous solution to form higher ordered multimeric structures, which are very reminiscent of natural plant storage proteins. The DNA encoding this amino acid sequence was synthesized, and from this monomeric gene fragment (heaae-1), the stable tetrameric form of the gene (heaae-4) was generated by subcloning into the E. coli expression vector pKK223-3. A clear 6 kDa polypeptide band corresponding to the molecular weight of the dimeric form (HEAAE-2) was detected. The smeared band which appeared around the molecular weight corresponding to HEAAE-4 of 11 kDa suggested that the tetramer form of this protein might be processed into smaller size products.

• Molecules and Cells
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• v.45 no.7
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• pp.495-501
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• 2022

Leucine dehydrogenase (LDH, EC 1.4.1.9) catalyzes the reversible deamination of branched-chain L-amino acids to their corresponding keto acids using NAD⁺ as a cofactor. LDH generally adopts an octameric structure with D4 symmetry, generating a molecular mass of approximately 400 kDa. Here, the crystal structure of the LDH from Pseudomonas aeruginosa (Pa-LDH) was determined at 2.5 Å resolution. Interestingly, the crystal structure shows that the enzyme exists as a dimer with C2 symmetry in a crystal lattice. The dimeric structure was also observed in solution using multiangle light scattering coupled with size-exclusion chromatography. The enzyme assay revealed that the specific activity was maximal at 60℃ and pH 8.5. The kinetic parameters for three different amino acid and the cofactor (NAD⁺) were determined. The crystal structure represents that the subunit has more compact structure than homologs' structure. In addition, the crystal structure along with sequence alignments indicates a set of non-conserved arginine residues which are important in stability. Subsequent mutation analysis for those residues revealed that the enzyme activity reduced to one third of the wild type. These results provide structural and biochemical insights for its future studies on its application for industrial purposes.

• Journal of the Korean Wood Science and Technology
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• v.31 no.6
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• pp.31-39
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• 2003

The enzymatic isolation of residual lignins obtained from spruce and beech pulps (obtained by sulfite, kraft, ASAM and soda/AQ/MeOH pulping processes) and their characterization was described in previous publications. Here, the residual lignins have been submitted to potassium permanganate oxidation (KMnO₄ degradation), and 9 aromatic carboxylic acids (3 of them are dimeric) were identified after methylation with diazomethane by GC/MS. The analytical challenge during qantification by the internal standard methods was the partly high protein content of the samples, which resulted in elevated anisic acid yields in the degradation mixture of sulfite residual lignins. The results are compared with the KMnO₄ degradation of the corresponding MWLs and discussed in terms of S/G ratios and degrees of condensation. The latter was calculated as a quotient between the aromatic carboxylic acids derived from condensed and non-condensed lignin structures. Typical degradation patterns for the various processes have been observed. Among other parameter, the relative compositions between iso-hemipinic acid (which is for condensation in pos. 5 of the aromatic ring) and meta-hemipinic acid and 3,4,5-trimethoxyphthalic acid (both are for condensation in pos. 6 of the aromatic ring) was found to be process specific. Kraft and soda/AQ/MeOH residual lignins yielded higher amounts of iso-hemipinic acid. In contrast, the relative yields of meta-hemipinic acid and 3,4,5-trimethoxyphthalic acid (the latter in beech lignins) are higher in sulfite and particularly in ASAM residual lignin. In case of beech residual lignins the amount of acids originated from non-condensed syringyl type lignin units was surprisingly high. The condensation degree of residual lignins was shown to be generally higher than that of MWLs. This was especially true for the G units. ASAM residual lignin exhibited very high S/G ratios and degrees of polymerization. Causality between condensation degree and total yield of degradation products was demonstrated.

• BMB Reports
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• v.45 no.4
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• pp.239-241
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• 2012

Iron availability is limited in the environment and most bacteria have developed a system to acquire iron from host hemoproteins. Heme oxygenase plays an important role by degrading heme group and releasing the essential nutrient iron. The structure of Bacillus subtilis HmoB was determined to 2.0 ${\AA}$ resolution. B. subtilis HmoB contains a typical antibiotic biosynthesis monooxygenase (ABM) domain that spans from 71 to 146 residues and belongs to the IsdG family heme oxygenases. Comparison of HmoB and IsdG family proteins showed that the C-terminal region of HmoB has similar sequence and structure to IsdG family proteins and contains conserved critical residues for heme degradation. However, HmoB is distinct from other IsdG family proteins in that HmoB is about 60 amino acids longer in the N-terminus and does not form a dimer whereas previously studied IsdG family heme oxygenases form functional homodimers. Interestingly, the structure of monomeric HmoB resembles the dimeric structure of IsdG family proteins. Hence, B. subtilis HmoB is a heme oxygenase with a novel structural feature.

• BMB Reports
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• v.41 no.11
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• pp.790-795
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• 2008

An inducible lysine 6-dehydrogenase (Lys 6-DH), which catalyzes the oxidative deamination of the 6-amino group of L-lysine in the presence of $NAD^+$, was purified to homogeneity from Achromobacter denitrificans, yielding a homodimeric protein of 80 kDa. The enzyme was specific for the substrate L-lysine and $NAD^+$ served as a cofactor. The dimeric enzyme associated into a hexamer in the presence of 10 mM L-lysine. The $K_m$ values for L-lysine and $NAD^+$ were 5.0 and 0.09 mM, respectively. The lys 6-dh gene was cloned and overexpressed in E. coli. The open reading frame was 1,107 nucleotides long and encoded a peptide containing 368 amino acids with 39,355 Da. The recombinant enzyme was purified to homogeneity and characterized. Enzyme activities and kinetic properties of the recombinant enzyme were almost the same as those of the endogenous enzyme obtained from A. denitrificans. Crystals of the enzyme were obtained using the hanging drop method.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.327-333
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• 2005

Dihydroxy-acid dehydratase (DHAD, 2,3-dihydroxy-acid hydrolyase, EC 4.2.1.9) is one of the key enzymes involved in the biosynthetic pathway of the branched chain amino acid isoleucine and valine. Although the enzyme have been purified and characterized in various mesophiles including bacteria and eukarya, the biochemical properties of DHAD has bee not yet reported from hyperthermophilic archaea. In this study, we cloned, expressed, and purified a DHAD homologue from the thermoacidophilic archaeon Sulfolobus solfataricus P2, which grows optimally at $80\;^{\circ}C$ and pH 3, in E. coli. Characterization of the recombinant S. solfataricus DHAD (rSso_DHAD) revealed that it is the dimeric protein with a subunit molecular weight of 64,000 Da in native structure. rDHAD showed the highest activity toward 2,3-dihydroxyisovaleric acid among 17 aldonic acid substrates Interestingly, this enzyme also displayed 50 % activities toward some pentonic acids and hexonic acids when compared with the activity of this enzyme to the natural substrate. Moreover, rSso_DHAD indicated relatively higher activity toward D-gluconate than any other hexonic acids tested in substrates. $K_m$ and $V_{max}$ values of rSso_DHAD were calculated as $0.54\;{\pm}\;0.04\;mM$ toward 2,3dihydroxyisovalerate and $2.42\;{\pm}\;0.19\;mM$ toward D-gluconate, and as $21.6\;{\pm}\;0.4\;U/mg$ toward 2,3-dihydroxyisovalerate and $13.8\;{\pm}\;0.4\;U/mg$ toward D-gluconate, respectively. In the study for biochemical properties, the enzyme shows maximal activity between $70^{\circ}C$ and $80^{\circ}C$, and the pH range of pH 7.5 to 8.5. The half life time at $80^{\circ}C$ was 30 min. A divalent metal ion, $Mn^{2+}$, was only powerful activators, whereas other metal ions made the enzyme activity reduced. $Hg^{2+}$, organic mercury, and EDTA also strongly inhibited enzyme activities. Particularly, the rSso_DHAD activity was very stable under aerobic condition although the counterparts reported from mesophiles had been deactivated by oxygen.

• Korean Journal of Food Science and Technology
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• v.5 no.1
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• pp.36-41
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• 1973

Chemical changes with time of frying-fats during the frying process (beef tallow with 0.01% BHA and 0.01% BHT, turnover rate 9%/hr and $140{\pm}10^{\circ}C$ temperature) of Ramyon on a commercial scale were studied. No significant changes of carbonyl value and peroxide value were noted up to 120 hrs. However, small increases in acid value, color and dimeric fatty acid were noted. Small decreases in iodine value and the content of unsaturated fatty acids were also observed. Under the experimental storage condition, when the stability of fats heated were compared to fresh fat, minor differences in carbonyl value and weight gain were noted. All of these demonstrated that frying-fats commercially used in Ramyon frying system were maintained in good quality during the frying process.