• Journal of Sericultural and Entomological Science
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• v.39 no.2
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• pp.169-173
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• 1997

The behavior of tyrosine(Tyr.) residue of Bombyx mori silk fiber from yellow fluorescent cocoon has been examined for the dependence of pH in aqueous silk solution under the presence of orange II salt. Through the peak separation of angular dependence of spectral pattern of 15N-Tyr. and [1-13C]-Tyr. between the fiber axis and the molecular bond direction, N-H bond in fiber as well as the orientation distribution around the fiber axis were analyzed. Also, and sericin component was obtained from these angular dependence of oriented spectral pattern. The pH dependence of the 13C NMR chemical shift of B. mori silk fibroin was examined in aqueous solution in the presince of orange II are broad at pH$\geq$7.0. However, these become sharper at pH$\geq$8.0 and remain sharp at higher pH. In these higher pH range, a chemical shift change occurs due to the deprotonation of the Tyr. side group of fibroin. At higher pH. such a hydrophobic cluster is destroyed because of the electrostatic interaction according to the deprotonation of the Tyr-OH group.

• Bulletin of the Korean Chemical Society
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• v.24 no.8
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• pp.1188-1192
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• 2003

In order to study the role of residue in the active site of glutathione S-transferase (GST), Tyr 108 residue in human GST P1-1 was replaced with alanine, phenylalanine and tryptophan by site-directed mutagenesis to obtain mutants Y108A, Y108F and Y108W. These three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitutions of Tyr108 significantly affected $K_m^{CDNB}$ and $K_m^{ETA}$, whereas scarcely affected $K_m^{GSH}$. The substitutions of Tyr108 also significantly affected $I_{50}$ of ETA, an electrophilic substrate-like compound. The effect of these substitutions on kinetic parameters and the response to inhibition suggests that tyrosine 108 in hGST P1-1 contributes to the binding of the electrophilic substrate and a major determinant in the binding of CDNB is the aromatic ring of Tyr108, not its hydroxyl group.

• Preventive Nutrition and Food Science
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• v.7 no.4
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• pp.397-404
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• 2002

Soybean curd residue (SCR) was fermented by lactic acid bacteria, Lactobacillus rhamnosus LS and Entercoccus faecium LL, isolated from SCR. The pH, titratable acidify and viable cell counts were determined from the fermented SCR to evaluate the lactic acid production and growth of lactic acid bacteria. Optimal amounts of pretense enzyme and glucose, and ideal fermentation time for SCR fermentation were estimated by response surface methodology (RSM). Raw SCR fermented by indigenous microorganisms had 0.78 % titratable acidity, The acid production in SCR fermented by L. rhamnosus LS was greatly enhanced by the addition of glucose and lactose. However only glucose increased acid production by Ent. faecium LL. The proof test of SCR fermentation demonstrated that similar results for titratable acidity, tyrosine content and viable cell counts to that predicted could be obtained by the at optimized fermentation conditions. In the presence of 0.029 % (w/w) pretense enzyme and 0.9% (w/w) glucose, the SCR fermented by Ent. faecium LL showed 1.07% (w/v) of titratable acidity, 1.02 mg% tyrosine content and 2$\times$10$^{9}$ (cfu/g) of viable cell counts. With the SCR fortified with 0.033% pretense enzyme and 1.7% glucose, L. rhamnosus LS showed 1.8% (w/v) of titratable acidity, 0.92 mg% of tyrosine content and 2$\times$10$^{9}$ (cfu/g) of viable cell counts.

• Bulletin of the Korean Chemical Society
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• v.34 no.9
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• pp.2671-2674
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• 2013

To elucidate the role of the evolutionarily conserved Tyr8 residue in rice Phi-class GSTF3, this amino acid was replaced with alanine and phenylalanine by site-directed mutagenesis, respectively. The replacement of Tyr8 with Ala significantly affected the catalytic activity and the kinetic parameters, whereas the substitutions of Tyr8 with Phe had almost no effect. The Y8A mutant resulted in approximately 90-100% decrease of the specific activity. Moreover, the Y8A mutant resulted approximately in 2-fold increase of $K_m$, approximately 60-80% decrease of $k_{cat}$, and approximately 6.5-fold decrease in $k_{cat}/K_m$. From the pH/log $k_{cat}/K_m$ plot, $pK_a$ values of the GSH in the wild-type enzyme-GSH complex, Y8A-GSH complex and Y8F-GSH complex were estimated to be approximately 6.8, 8.5 and 6.9, respectively. From these results, we suggest that the evolutionarily conserved Tyr8 residue in OsGSTF3 seems to influence the structural stability of the active site of OsGSTF3 rather than directly its catalytic activity.

• Preventive Nutrition and Food Science
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• v.9 no.2
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• pp.138-143
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• 2004

Soybean curd residues (SCR) obtained from hot and cold manufacturing processes were fermented by indigenous microorganisms, Lactobacillus rhamnosus LS and Bacillus firmus NA-l for 15 h at 37$^{\circ}C$. The pH, acidity, viable cell counts, and tyrosine content were evaluated in samples with variations in sugar, starter and type of SCR. The raw Doowon SCR (D-SCR, cold-processed) fermented by indigenous microorganism had a 0.9% acidity and 6.7 ${\times}$ 10$^{7}$ CFU/g viable cell counts, compared with the 0.11 % acidity and 6.7 ${\times}$ 10$^{6}$ CFU/g viable cell counts of raw fermented Pulmuwon SCR (P-SCR, hot-processed). After fermentation of raw P-SCR with 1 % glucose and 1 % L. rhamnosus LS starter, the viable cell counts, tyrosine content and acidity were 4.7 ${\times}$ 10$^{8}$ CFU/g, 16.3 mg% and 0.9%, respectively. In addition, the raw P-SCR fermented with Bacillus firmus NA-l as co-starter had a 0.45% acidity, 2.4 ${\times}$ 10$^{8}$ CFU/g lactic acid bacteria, and 3.3 ${\times}$ 10$^{6}$ CFU/g Bacillus sp. In particular, the tyrosine content was increased 5 fold. The drying of fermented SCR was completed by hot-air drying (5$0^{\circ}C$) within 12 h; the dried P-SCR and D-SCR had 1.8 ${\times}$ 10$^{7}$ CFU/g and 5.3 ${\times}$ 10$^{6}$ CFU/g viable cell counts, respectively. The concentrate of methanol extract from fermented D-SCR inhibited the initial cell growth of E. coli, Staphylococcus aureus and Pseudomonas aeruginosa in liquid culture.

• Bulletin of the Korean Chemical Society
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• v.35 no.8
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• pp.2299-2303
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• 2014

Two diastereoisomers of cyclo(Pro-Tyr) have been synthesized simultaneously. The crystal structures and conformations of both cyclo($\small{L}$-Pro-$\small{L}$-Tyr) and a racemic mixture of cyclo($\small{D}$-Pro-$\small{L}$-Tyr) and cyclo($\small{L}$-Pro-$\small{D}$-Tyr), abbreviated as rac-cyclo($\small{D}$-Pro-$\small{L}$-Tyr/$\small{L}$-Pro-$\small{D}$-Tyr), have been determined by a single-crystal X-ray diffraction study at low temperature. The crystals of rac-cyclo($\small{D}$-Pro-$\small{L}$-Tyr/$\small{L}$-Pro-$\small{D}$-Tyr) belong to orthorhombic space group $Pna2_1$ with a = 10.755 (1), b = 12.699 (1), c = 9.600 (1) ${\AA}$ and Z = 4. The tyrosine side chain is folded towards the diketopiperazine (DKP) ring. The DKP ring adopts a twist boat conformation with pseudo symmetry $C_{2v}$. The pyrrolidine ring has an envelope conformation with the N5, C4, C7 and C8 atoms in a plane. The crystal of rac-cyclo($\small{D}$-Pro-$\small{L}$-Tyr/$\small{L}$-Pro-$\small{D}$-Tyr) is stabilized by hydrogen bonds between amide N2-H2 and carbonyl oxygen O2 in the neighbor. The hydroxyl group of tyrosine residue is also hydrogen bonded to the oxygen of the carbonyl group of the DKP ring in the next molecule. The spectroscopic properties of both isomers are also described.

• BMB Reports
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• v.28 no.4
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• pp.293-300
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• 1995

The radical centers detected in the reaction of metmyoglobin (MetMb) with hydrogen peroxide ($H_2O_2$) have been studied by using a spin trapping technique. A broad 5-line asymmetric electron spin resonance (ESR) spectrum, with $2A_{max}=4.07\;mT$ and $2A_{min}=2.97\;mT$, obtained after incubation of MetMb with $H_2O_2$ in the presence of a spin trap, 5,5-dimethyl pyrroline-N-oxide (DMPO) was gradually weakened with time and disappeared completely by 6 min after addition of guanidine-HCl (14 M). When a higher concentration (6 M) of the agent was added, the signal disappeared within 40 see and the DMPO/OH signal appeared immediately. Then, a new 8-line signal with similar intensities grew gradually and was fixed by 45 min, coexisting with the DMPO/OH signal. This new signal was found to be composite, consisting of two different radical species. One of the 6-line signals, with $a_N$ 1.49 mT and $a_H$ 0.988 mT, was assigned to the DMPO/phenoxyl radical adduct. The second 6-line signal with $a_N$ 1.55 mT and $a_H$ 2.22 mT was assigned to carbon-centered radical adduct. When 3,3,5,5-tetramethylpyrrolin-N-oxide (TMPO), was employed in the place of DMPO, another broad asymmetric 5-line signal was detected with $2A_{max}=3.99\;mT$ and $2A_{min}=3.04\;mT$, which is virtually identical to that obtained from the DMPO system The shape of the spectrum of the TMPO adduct changed drastically, with lapse of time resulting in a broad singlet after 40 min. The broad singlet was assigned to the porphyrin radical adduct. Incubation of globin with Fenton reagent in the presence of DMPO initially gave a DMPO/OH signal. Then, a new 12-line signal began to grow after one minute and fixed after 15 min. coexisting with the DMPO/OH signal, This 12-line signal was assigned to DMPO/phenoxyl with $a_N$ 1.47 mT, $a_{{\beta}H}$ 0.99 mT and $a_{{\gamma}H}$ 0.13 mT. A minor concentration of carbon-centered radical adduct was also detected. This radical composition is identical to that of guanidine HCl treated MetMb/DMPO/$H_2O_2$ system, indicating that the radical producing conditions are somehow common in both systems. Heme iron can be released by excess $H_2O_2$ in the MetMb/$H_2O_2$ system, providing for Fenton reagent. When MetMb was pretreated with tyrosine blocking agent, $KI_3$ the broad 5.line MetMb-derived signal was not detected in the MetMb/DMPO/$H_2O_2$ system, whereas no such effect was detected on such system of Hb in which the radical center was assigned to cysteine residue not tyrosine, indicating that tyrosine residue is a main radical center produced in the MetMb/$H_2O_2$ system Thus, the present data strongly support the previous indication that the apomyoglobin-derived radical center formed in the reaction of MetMb with $H_2O_2$ is a tyrosine residue.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.551-558
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• 1992

The extracellular endoinulase from Streptomyces sp. 556 was purified and characterized, The culture broth was fractionated by ammonium sulfate saturation followed by DEAE-cellulose column chromatography and 5ephadex G-200 gel filtration, The ultimately purified fraction revealed a single band in 7.5% polyacrylamide gel electropherogram. The purified enzyme showed the maximal activity at pH 5.5-6.0 and $50^{\circ}C$, but lost 93% of inulase activity after 30 min incubation at $55^{\circ}C$ . The essen.tial amino acid residue for catalytic activity appeared to be tryptophan. This endo inulase was activated by $Mn^{2+}$, whereas inactivated by $Ag^{+}$, $Hg^{+}$, $Cu^{2+}$, $Zn^{2+}$, $Fe^{3+}$ and $Mo^{6+}$ EDTA and 8-hydroxyquinoline inhibited the enzyme so that the enzyme was considered to be a metalloenzyme. The Km value for inulin was 0.287 mM, and no invertase or $\alpha$-glucosidase activity was found in the enzyme.

• The Korean Journal of Zoology
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• v.33 no.4
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• pp.468-475
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• 1990

An In vitro protein sulfation in the soluble fraction of rat brain was charaderized further by an improved method of alkaline hydrolysis and thin layer ceflulose electrophoresis TLE) The protein sulfation was carried out in a reaction system containing [35 S] 3'-phosphoadenosine-5'-phosphosulfate (PAPS), Tris-maleate buffer (pH 8), MgCI$_2$, and soluble proteins from rat brain. The sulfated proteins were precipitated by acetone and alkaline hydrolysis was performed to obtain sulfated amino acids. The hydrolysate was separated further by TLE and the separated residues were identified by fluorography. The Iluorography of one-dimensional The showed at least nine sulfated residues including tryosine-O-sulfate. The other spots were not identified yet positively. General properties of protein sulfotransferases (PST) using this method were re-examined such as effects of concentrations of PAPS, pH, incubation temperature and $Mg^2$+. These results suggest a possible occurrence of several PST corresponding to each sulfated residue in rat brain and that the sulfation can occur not only in tyrosine but also in other residues as well.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.501-519
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• 2003

Silk Sericin(SS) is a natural protein extracted from cocoon of bombix mori and shows moisturizing effect to the skin due to a number of hydroxyl groups in the structure. But its application to cosmetics is limited due to its poor solubility in water. In order to solve this drawback and expand its application to cosmetics, polyethyleneglycol(PEG) was conjugated with sericin by reacting activated polyethyleneglycol(ActPEG). Reaction site of sericin is tyrosine residue, which was determined by using $^1$H-NMR. Random coil structure of sericin was transformed to beta-sheet structure by conjugating polyethyleneglycol. It was confirmed that melting point of sericin-PEG conjugate was lowered compared to that of each sericin and PEG due to the interaction between sericin and PEG in crystalline structure. Self-assembled sericin-PEG nanoparticle was obtained by dialyzing with alcohol solution of sericin-PEG conjugate against water. The particle is spherical and has 200-400nm of size. The moisturizing ability of sericin-PEG nanoparticle was much higher than that of sericin itself. Incorporation of vitamin A into sericin-PEG nanoparticle was carried out by diafiltration method. The content of incorporated Vitamin A in sericin-PEG nanoparticle was 8.9 wt％. Releasing behaviour of vitamin A incorporated into nanoparticle was tested in phosphate buffer, pH 7.4 at 37$^{\circ}C$. and half-life of Vitamin A release was 43hrs. Sericin-PEG nanoparticle exhibited higher moisturing effect than sericin itself and distilled water, respectively. No toxicity and irritation were observed in animal tests. It can be expected that the self-assembled sericin-PEG nanoparticle can be developed for cosmetics.