• BMB Reports
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• v.38 no.1
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• pp.115-119
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• 2005

Replacement of valine by tryptophan or tyrosine at position $\alpha$96 of the $\alpha$ chain ($\alpha$96Val), located in the ${\alpha}_1{\beta}_2$ subunit interface of hemoglobin leads to low oxygen affinity hemoglobin, and has been suggested to be due to the extra stability introduced by an aromatic amino acid at the $\alpha$96 position. The characteristic of aromatic amino acid substitution at the $\alpha$96 of hemoglobin has been further investigated by producing double mutant r Hb ($\alpha$42Tyr$\rightarrow$ Phe, $\alpha$96Val$\rightarrow$Trp). r Hb ($\alpha$42Tyr$\rightarrow$Phe) is known to exhibit almost no cooperativity in binding oxygen, and possesses high oxygen affinity due to the disruption of the hydrogen bond between $\alpha$42Tyr and $\beta$99Asp in the ${\alpha}_1{\beta}_2$ subunit interface of deoxy Hb A. The second mutation, $\alpha$96Val$\rightarrow$Trp, may compensate the functional defects of r Hb ($\alpha$42Tyr$\rightarrow$Phe), if the stability due to the introduction of trypophan at the $\alpha$96 position is strong enough to overcome the defect of r Hb ($\alpha$42Tyr$\rightarrow$Phe). Double mutant r Hb ($\alpha$42Tyr$\rightarrow$Phe, $\alpha$96Val$\rightarrow$Trp) exhibited almost no cooperativity in binding oxygen and possessed high oxygen affinity, similarly to that of r Hb ($\alpha$42Tyr$\rightarrow$Phe). $^1$H NMR spectroscopic data of r Hb ($\alpha$42Tyr$\rightarrow$Phe, $\alpha$96Val$\rightarrow$Trp) also showed a very unstable deoxy-quaternary structure. The present investigation has demonstrated that the presence of the crucible hydrogen bond between $\alpha$42Tyr and $\beta$99Asp is essential for the novel oxygen binding properties of deoxy Hb ($\alpha$96Val$\rightarrow$Trp).

• Journal of Life Science
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• v.28 no.2
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• pp.195-200
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• 2018

Beta-asarone is the well-known active ingredient of Rhizoma acori graminei. In this study, we investigated and compared the binding affinity of mosquito oviposition pheromone (MOP; (5R,6S)-6-acetoxy-5-hexadecanolide) and beta-asarone on the A domain of the mosquito odorant binding protein 1 (CquiOBP1) by in silico computational docking studies. The three-dimensional crystallographic structure of CquiOBP1 was obtained from the PDB database (PDB ID: 3OGN). In silico computational auto-docking analysis was performed using PyRx, Autodock Vina, Discovery Studio Version 4.5, and the NX-QuickPharm option based on scoring functions. The beta-asarone showed optimum binding affinity (docking energy) with CquiOBP1 as -6.40 kcal/mol as compared to the MOP (-6.00 kcal/mol). Among the interacting amino acids (LEU76, LEU80, ALA88, MET89, HIS111, TRP114, and TYR122), tryptophan 114 in the CquiOBP1 active site significantly interacted with both MOP and beta-asarone. Amino acids substitution (mutation) from non-polar groups to the polar (or charged) groups of the CquiOBP1 dramatically changed the X, Y, Z grid position and binding affinity of both ligands. These results significantly indicated that beta-asarone could be a more potent ligand to the CquiOBP1 than MOP. Therefore, the extract of Rhizoma acori graminei or beta-asarone can be applied to the fields of insecticidal and repellant biomaterial development.

• Bulletin of the Korean Chemical Society
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• v.32 no.8
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• pp.2577-2583
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• 2011

We have designed a 20-residue hybrid peptide CA(1-8)-MA(1-12) (CAMA) incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA) with high bacterial cell selectivity. CAMA-P2 is an ${\alpha}$-helical antimicrobial peptide designed from a CAMA hybrid peptide and substitution of Gly-Ile-Gly hinge sequence of CAMA to Pro influences the flexibility at central part of CAMA. Based on structure-activity relationships of CAMA peptides, to investigate the effects of the total positive charges on antimicrobial activity of CAMA-P2, the $Ser^{14}{\rightarrow}$Lys analogue (CAMA-syn1) was synthesized. The role of tryptophan at C-terminal ${\alpha}$-helix on its antimicrobial activity as well as synergistic activity was also investigated using $Ser^{14}{\rightarrow}$Lys/$Phe^{18}{\rightarrow}$Trp analogue (CAMA-syn2). Also, we designed CAMA-syn3 by substitution of $Lys^{16}$ located opposite side of substituted $Lys^{14}$ of CAMA-syn1 with Leu residue, resulting in increase of hydrophobicity and amphipathicity of the peptide. All of CAMA-syn analogues showed good antimicrobial activities similar to those of CAMA and CAMA-P2. The CAMA-syn1 and CAMA-syn2 showed low hemolytic activity and cytotoxicity against human keratinocyte Haca-T cells while CAMA-syn3 showed hemolytic activity and cytotoxicity at its MIC value. We then investigated their abilities to act synergistically in combination with the antimicrobial flavonoids and synthetic compounds screened in our laboratory. The results showed that all peptides exhibited synergistic effects with dihydrobinetin, while only CAMA-syn2 exhibited synergistic effects with YKAs3001 against both S. aureus and MRSA, suggesting that Trp residue at C-terminus of CAMA-syn2 may facilitate the polar antibiotic flavonoids and synthetic compounds to permeabilize the membrane. This study will be useful for the development of new antibiotic peptides with potent antimicrobial and synergistic activity but without cytotoxicity.

• Biomedical Science Letters
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• v.13 no.2
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• pp.105-110
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• 2007

Aspartyl-tRNA synthetase (AspRS) exists in two different forms with respect to tRNA recognition. The discriminating enzyme (D-AspRS) recognizes only tRNA$^{Asp}$, while the non-discriminating one (ND-AspRS) also recognizes tRNA$^{Asn}$ and therefore forms both Asp-tRNA$^{Asn}$ and Asp-tRNA$^{Asp}$. Plus primary sequence distinguishes two general groups of AspRS. There is a predominantly bacterial-type, larger AspRS (about 580 aa) in addition to a shorter archaeal/eukaryotic type (about 430 aa). In vivo data made clear that discriminating and non-discriminating enzymes exist in both groups. The determinants in the protein sequence responsible for tRNA discrimination are not hewn. The AspRS from Acidithiobacillus ferrooxidans might be suggested ND-AspRS fur missing of AsnRS in genomic sequencing data. Therefore, we analyzed the AspRS from A. ferrooxidans with in vitro aminoacylation assay with E. coli unfractionated tRNA, in vivo missense suppression assay with tipA34 mutant and Northern hybridization with probes which were specific with tRNA$^{Asp}$ or tRNA$^{Asn}$. The AspRS from A. ferrooxidans produced more Asp-tRNA than that from E. coli. Only aspS gene from A. ferrooxidans suppressed trpA34 strain in minimal media without tryptophan. Only AspRS from A. ferrooxidans showed mischarged Asp-tRNA$^{Asn}$ band. Therefore, AspRS from A. ferrooxidans is definitely ND-AspRS.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.409-413
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• 2012

Indolicidin (ID) is a 13-residue Trp-rich antimicrobial peptide (AMP) isolated from bovine neutrophils. In addition to having a high antimicrobial potency, it is also toxic to mammalian cells. To develop novel ID-derived AMPs with shorter lengths and enhanced prokaryotic selectivities (meaning potent antimicrobial activity against bacterial cells without toxicity against mammalian cells) over the parental ID, several ID analogs were designed and synthesized. Finally, 10-residue ID analogs (SI, SI-PA, SI-WF and SI-WL) with much higher prokaryotic selectivity than the parental ID were developed. Our results suggest that the hydrophobic and aromatic amino acids at the central position of the analog SI with the highest prokaryotic selectivity are important for potent antimicrobial activity, but two Pro residues do not affect antimicrobial activity. The order of prokaryotic selectivity for ID and its designed analogs was SI > SI-PA > SI-WF > SI-WL > ID > SI-WA. Taken together, our designed short ID analogs could be developed as therapeutic agents for treating bacterial infections.

• Journal of the Korean Chemical Society
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• v.41 no.6
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• pp.304-312
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• 1997

Acetolactate synthase(ALS) is the common enzyme in the biosynthetic of valine, leucine, and isoleucine, and is the target of several classes of structually unrelated herbicides, including sulfonylureas, imidazolinones, and triazolopyrimidines. In an effort to develop new and desirable herbicides, we have synthesized 4,6-dimethoxypyrimidine derivatives, and examined their inhibitory activities on pea ALS. The most active compound was found to be K11570 and $IC_{50}$ value for K11570 was 0.2 ${\mu}M.$ The inhibition of pea ALS by K11570 was biphasic, showing increased inhibition with incubation time. The K11570 showed mixed-type inhibition with respect to substrate pyruvate. Dual inhibition analysis of K11570 versus sufonylurea herbicide Ally and feedback inhibitor leucine revealed that three inhibitors were competitive for binding to ALS. The arginine modified enzyme showed decreased inhibition by K11570, sufonylurea Ally, and leucine, in constrast to, tryptophan modification did not affect on the sensitivity of ALS to the inhibitors.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.305-311
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• 2001

Antifungal bacterium, Bacillus subtilis YS1 was isolated from Yusong hot spring showed broad antifungal spectrum against 12 kinds of plant pathogenic fungi and Candida albicans, animal pathogen. From the gamma($Co^{60}$) radiation sensitivity test, $D_10$ value was 2.08 kGy and it survived above 20 kGy of radiation dose. Several mutants were induced by gamma radiation. Among them, YS1-1009 mutant showed resistance against tebuconazole of herbicide, increased activity against Botryoshaeria dothidea and ligninase activity. YS67 mutant was antifungal deficient auxotrophic mutants(trp-pro-or arg-ura-). From this results, it suggested that gamma irradiation could be useful method for mutant induction.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.201-208
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• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• Animal Bioscience
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• v.36 no.10
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• pp.1584-1595
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• 2023

Objective: This study was conducted to evaluate the effect of a low-protein diet on growth performance, carcass traits, nutrient digestibility, blood profiles, and odor emissions in growing-finishing pigs. Methods: A total of 126 crossbred pigs ([Yorkshire×Landrace]×Duroc) with an average body weight (BW) of 38.56±0.53 kg were used for a 14-week feeding trial. Experimental pigs were allotted to one of 6 treatments in 3 replicates of 7 pigs per pen in a randomized complete block design. Pigs were fed each treatment diet with different levels of crude protein (CP). Phase 1 (early growing): 14%, 15%, 16%, 17%, 18%, 19%; phase 2 (late growing): 13%, 14%, 15%, 16%, 17%, 18%; phase 3 (early finishing): 12%, 13%, 14%, 15%, 16%, 17%; phase 4 (late finishing): 11%, 12%, 13%, 14%, 15%, 16%. All experimental diets in each phase were contained the same concentration of lysine (Lys), methionine (Met), threonine (Thr), and tryptophan (Trp). Results: Over the entire experimental period, there was no significant difference in BW, average daily feed intake, and gain-to-feed ratio among all treatments (p>0.05), but a quadratic effect (p = 0.04) was observed in average daily gain (ADG) during the late finishing phase with higher ADG in Group D. Blood urea nitrogen concentration linearly increased with an increase in dietary CP levels (p<0.01). Regarding nutrient digestibility, excreted nitrogen in urine and feces and nitrogen retention linearly increased as the CP level increased (p<0.01). A linear effect was observed with increasing CP levels in amines, ammonia, and hydrogen sulfide in odor emissions (p<0.01). No significant effects were observed in the measurements of carcass traits and meat characteristics (p>0.05). Conclusion: In phase feeding, reducing the CP level to 14% in early-growing pigs, 13% in late-growing pigs, 12% in early-finishing pigs, and 11% in late-finishing pigs is recommended.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.5
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• pp.655-667
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• 2001

This experiment was conducted to investigate the effect of feeding a low CP diet supplemented with synthetic amino acids on performance, nutrient utilization and carcass characteristics of finishing pigs fed under a three-phase feeding regimen. Ninety-six finishing pigs (Landrace$\times$Large White$\times$Duroc), $55.75kg{\pm}0.65$ of initial body weight, were blocked by weight and sex and allotted to four dietary treatments in a randomized block design. There were six pens per treatment and four pigs per pen. Pigs were fed a 16%-14%-12% CP (for phase I-II-III, respectively), sequence of diets. Dietary treatments were 1) Control, 2) Con+L (a sequence of diets reduced in CP by l percentage unit with lysine (L) supplementation, 3) Con+LMT (a sequence of diets reduced in CP by 2 percentage unit with LYS, methionine (MET) and threonine (THE) supplementation) and 4) Con+LMTT (a sequence of diets reduced in CP by 3 percentage unit with LYS, MET, THR and tryptophan (TRP) supplementation). The finishing period (55 to 105 kg) was divided into three phases (55 to 72 kg, 72 to 90 kg and 90 to 105 kg). Pigs fed either the control or Con+L diet grew faster (p<0.05) than pigs fed the Con-LMT or Con+LMTT diet. There was no difference in ADFI among dietary treatments. Phosphorus (P) digestibility was lowest in the control group and highest in the Con+LMTT group (p<0.05). Within each phase, no significant differences in dry matter (DM) and CP digestibilities were found. Although some amino acid digestibilities were affected by dietary treatments, digestibilities of essential amino acids (EAA), non-essential amino acids (NEAA) and total amino acid were not significantly influenced by dietary treatments. For the entire experiment periods, Con+L, Con+LMT and Con+LMTT treatments resulted in 13.4, 18.8 and 21.6% lower total N excretion compared with the control. Con+LMT and Con+LMTT treatments showed significantly lower BUN concentration compared with the control and Con+L treatment (p<0.05), but there was no significant difference in BUN concentration between pigs fed the control and Con+L treatment or between pigs fed Con+LMT and Con+LMTT treatments (p>0.05). Carcass length, backfat thickness and carcass grade were not significantly affected by dietary treatments (p>0.05). In conclusion, reducing dietary CP level by 1 percentage unit and supplementing only LYS at each phase could be a very beneficial feeding strategy for finishing pigs fed under a three phase feeding regimen in terms of both environmental and economical aspects.