• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.296-300
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• 1990

For interspecific portoplast fusion, Brevibacterium flauum lOAHR (Rifr axg his) and Corynebacterium glutamicum 11TS ($Sm-r$ trp) were induced by UV and NTG treatment. The protoplast fusion frequency between E. flavum XOAHR and C. glutamicum llTS was $3.7\times 10^{-6}$ with the lysozyme treatment (300 P $\mu g$ml) for 18 hrs. Genotypes of recombinants were analized as FMM ($Rif^r\; Sm^r$), FA (Rift $Sm^r$ arg), FH ($Rif^r\; Sm^r$ his), FT ($Rif^r\; Sm^r$ trp), FAH ($Rif^r\; Sm^r$ arg trp), FAT ($Rif^r\; Sm^r$ arg trp), and FAHT ($Rif^r\; Sm^r$ arg his trp). FAH 1 produced 12 fold of glutamate production compared to parental type, E. flauum 10AHR. In glutamine productivity, it produced 2.6 fold to parental type, C. glutamicum 11TS. Production of glutamate or glutamine by recombinants was involved in the specific activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS), respectively.

• BMB Reports
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• v.54 no.10
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• pp.516-521
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• 2021

Although arginase primarily participates in the last reaction of the urea cycle, we have previously demonstrated that arginase II is an important cytosolic calcium regulator through spermine production in a p32-dependent manner. Here, we demonstrated that rhaponticin (RPT) is a novel medicinal-plant arginase inhibitor and investigated its mechanism of action on Ca²⁺-dependent endothelial nitric oxide synthase (eNOS) activation. RPT was uncompetitively inhibited for both arginases I and II prepared from mouse liver and kidney. It also inhibited arginase activity in both aorta and human umbilical vein endothelial cells (HUVECs). Using both microscope and FACS analyses, RPT treatments induced increases in cytosolic Ca²⁺ levels using Fluo-4 AM as a calcium indicator. Increased cytosolic Ca²⁺ elicited the phosphorylations of both CaMKII and eNOS Ser1177 in a time-dependent manner. RPT incubations also increased intracellular L-arginine (L-Arg) levels and activated the CaMKII/AMPK/Akt/eNOS signaling cascade in HUVECs. Treatment of L-Arg and ABH, arginase inhibitor, increased intracellular Ca²⁺ concentrations and activated CaMKII-dependent eNOS activation in ECs of WT mice, but, the effects were not observed in ECs of inositol triphosphate receptor type 1 knockout (IP3R1^-/-) mice. In the aortic endothelium of WT mice, RPT also augmented nitric oxide (NO) production and attenuated reactive oxygen species (ROS) generation. In a vascular tension assay using RPT-treated aortic tissue, cumulative vasorelaxant responses to acetylcholine (Ach) were enhanced, and phenylephrine (PE)-dependent vasoconstrictive responses were retarded, although sodium nitroprusside and KCl responses were not different. In this study, we present a novel mechanism for RPT, as an arginase inhibitor, to increase cytosolic Ca²⁺ concentration in a L-Arg-dependent manner and enhance endothelial function through eNOS activation.

• Journal of Animal Science and Technology
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• v.65 no.1
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• pp.69-95
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• 2023

In order to evaluate the effects of ARG sources (arginine [ARG] and Guanidinoacetic acid [GAA]) and phenylalanine (PHE) supplementation on performance, susceptibility to ascites, intestinal morphology, and nutrient digestibility in the cold-stressed broilers fed a canola meal (CM)-based diet, a 2×2 factorial experiment with four treatments was conducted. The dietary treatments included CM-based diet + 2.57 g/kg ARG, CM-based diet + 2.57 g/kg ARG + 1.5 g/kg PHE, CM-based diet + 1.8 g/kg GAA and CM-based diet + 1.8 g/kg GAA + 1.5 g/kg PHE. The corn-CM diet without supplementation was used as a negative control (NC) group in the fifth treatment that excluded the factorial arrangement. The results showed that adding ARG to diets without PHE supplement increased (p < 0.05) feed intake. Also, birds fed diets containing ARG had higher (p < 0.05) body weight gain (BWG) compared to those fed GAA added diets. Supplementation of PHE improved (p < 0.05) the FCR compared to groups fed diets without added PHE. Further, ARG addition increased (p < 0.05) plasma nitric oxide (NO) concentration, carcass, breast and leg yields, duodenal, jejunal, and ileal villus height (VH) to crypt depth (CD, and dry matter digestibility, while decreasing (p < 0.05) ascites mortality and right ventricle (RV) to total ventricle (TV) ratio compared to GAA added groups. Supplementation of PHE also declined susceptibility to ascites by reducing (p < 0.01) RV to TV ratio while increasing (p < 0.05) plasma NO level. The digestibility of ether extract also increased (p < 0.05) in broilers fed GAA supplemented diets versus those fed ARG added diets. The findings suggested that ARG may improve BWG and lower ascites incidence in broilers fed a diet based on CM under cold stress because of its antihypertensive effects. Moreover, the findings of this study demonstrated the importance of including PHE formulation in ARG-deficient diets to attenuate the adverse effects of cold stress on broilers. It was also concluded that GAA could be efficaciously used in cold-stressed broilers fed an ARG-deficient diet.

• Korea Journal of Cosmetic Science
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• v.2 no.1
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• pp.11-19
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• 2020

The aim of this study is to investigate the effect of various pentapeptides on hair repair depending on the characteristics of comprising amino acids using crosslinking agents in hair. Total ten peptides were synthesized with two kinds of amino acids respectively, of which were previously categorized according to R group of the amino acids contributing to the characteristic of each peptide: STTSS (Ser-Thr-Thr-Ser-Ser), LIILL (Leu-Ile-Ile-Leu-Leu), CMMCC (Cys-Met-Met-Cys-Cys), DEEDD (Asp-Glu-Glu-Asp-Asp), RKKRR (Arg-Lys-Lys-Arg-Arg), TAMRA-STTSS, TAMRA-LIILL, TAMRA-CMMCC, TAMRA-DEEDD, and TAMRA-RKKRR. Pentapeptide alone, or pentapeptides with crosslinking agents such as polymeric carbodiimide (PCI) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) were treated to chemically damaged hair. Hair diameter and break strength (N = 40/case) were measured to calculate tensile strength of hair for computing hair repair ratio, and fluorescence yields (N = 20/case) were collected for hair treated with TAMRA-peptides. The tensile strength of hair treated with pentapeptides alone, or pentapeptides with cross-linking agents is consistent with the fluorescence yield from the microscope images of the cross-sectioned hair in vision and in numerical values. Pentapeptides consisting of hydrophobic amino acids (LIILL), amino acids with sulfur (CMMCC), and basic amino acids (RKKRR) increased the tensile strength in perm-damaged hair. Pentapeptides with no extra carboxyl/amine groups in R group of amino acids resulted in no significant differences in hair strength and fluorescence yield among hairs treated with alone and with crosslinkers. Pentapeptides with extra carboxyl groups or amine groups enabled further strengthening of hair due to increased bonds within the hair after carbodiimide coupling reaction. The hair repairs of pentapeptides with various amino acid sequences were studied using crosslinking. Depending on the physical characteristics of comprising amino acids, the restoration of damaged hair was observed with tensile strength of hair and fluorescence signals upon cross-sectioned hair in parallel to possibly understand the binding tendency of each pentapeptide within the hair.

• Journal of Plant Biology
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• v.36 no.4
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• pp.391-398
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• 1993

It has been inferred that membrane-ECM (extracellular matrix) interaction in plants may be also mediated by an RGD-dependent recognition system as in animal cells. Effects of RGD peptide on ethylene production was examined in suspension cultured carrot cells. Treatment of the cells with RGD peptide containing RGD (Arg-Gly-Asp) sequence stimulated ethylene production. When RGD peptide was applied to carrot cells treated with 1M, the effect of RGD peptide appeared to be additive. ACC synthase activity in cells pretreated with RGD peptide likewise increased over the control. In an effort to check the sequence specificity of the RGD peptide, cells were treated with substituted RGD peptide, i.e. RGK (Arg-Gly-Lys) and RGE (Arg-Gly-Glu) peptide, respectively. RGK peptide did not stimulate ethylene production but RGE peptide did. The results strongly suggest that the stimulatory effect of RGD peptides on ethylene production may be associated with a physiological phenomenon through a specific recognition between RGD peptide including RGD sequence and their putative plasma membrane receptors.eptors.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.134-134
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• 2003

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.3
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• pp.279-284
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• 2001

This paper reports the purification of myomodulin A (MMA) and myomodulin E (MME) from the sea hare. The central nervous systems of 500 sea hare were extracted in an acidified solvent, after which four HPLC column systems were used to obtain pure peptides, The phasic contraction bioassay using a Mytilus edulis anterior byssus retractor muscle (ABRW) was applied to monitor all collected fractions. The pure peptides were submitted to Edman degradation based automated microsequencing. Mass spectrometry and chemical synthesis confirmed the sequence. The primary structures of MMA and MME were Pro-Met-Ser-Met-Leu-Arg-Leu-$NH_2$, (847.41 Da) and Cly-Leu-Gln-Met-Leu-Arg-Leu-$NH_2$, (830.50 Da), respectively. Synthetic peptides showed a modulating activity of phasic contraction in the ABRM of Mrtilus edulis.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.4
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• pp.301-305
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• 2001

The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 2002.05a
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• pp.215-215
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• 2002

Catalytic subunit of pyruvate dehydrogenase phosphatase (PDPc) has been suggested to have three major funational domains such as dihydrplipoamide adetyltransferase(E2)-binding domain, regulatory subunit of PDP(PDP)r-binding domain, and calcium-binding domain. In order to identify functional domains, recombinant catalytic subunit of pyruvate dehydrogenase phosphatase(rPDPc) was expressed in E. coli JM101 and purified to near homogeneity using the unique property of PDPc: PDPc binds to the inner lipoyl domain (L2) of E2 of ppyruvate dehydrogenase complex (PDC) in the presence of Ca+2, not under EGTA. PDPc was limited-proteolysed by typsin, chymotypsin, Arg-C, and elastase at pH 7.0 and 30C and N-terminal analysis of the fragments was done. Chymotrypsin, trypsin, and elastase made two major fragments: N-terminal large fragment, approx. 50kD and C-terminal small fragment, approx.10 kDa. Arg-C made three major fragments: N-terminal fragment, approx. 35kD, and central fragment, approx. 15 kD, and C-terminal fragment, approx. 10 kD. This study strongly suggest that PDPc consists of three major functional domains. However, further study should be necessary to identify the functional role.

• Environmental Analysis Health and Toxicology
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• v.17 no.1
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• pp.73-80
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• 2002

Pyruvate dehydrogenase phosphatase (PDP)와 kinase는 당대사시 해당과정에서의 대사 산물인 pyruvate를 acetyl CoA로 만들어 구연산 회로로 진입시켜 주는 효소인 pyruvate dehydrogenase complex (PDC)의 활성을 조절하는 중요한 효소이다. PDP의 catalytic subunit는 PDC의 dihydrolipoamide acetyltransferase (E2), PDP regulatory subunit (PDPr), 그리고 칼슘 결합 도메인 등으로 구성되어 있는 것으로 추측되어지고 있다. 본 연구에서는 그 구조와 기능과의 상관관계를 알아보기 위해 PDPc를 E. coli JM101에서 발현시켜 순수 정제 후 단백분해 효소를 이용한 제한적 가수분해 방법을 이용해 그 구조와 기능과의 상관관계에 대해 연구하고자 하였다 정제된 PDPc는 trypsin, chymotrypsin, Arg-C 그리고 elastase를 이용하여 3$0^{\circ}C$ 그리고 pH 7.0에서 제한적으로 분해시켰으며 각 분해산물의 아미노 말단의 아미노산 배열을 분석하였다. 그 결과 PDPc는 trypsin, chymotrypsin, elastase에 의해 N-terminal의 50 kD과 C-terminal의 10 kD의 두개의 분해산물을 만들었으며, Arg-C에 의해 50kD의 분해산물은 약 35kD와 15kD으로 더 가수분해가 되었다. 이러한 결과로 볼 때 PDPc는 앞에서 추측한데로 세개의 주요한 기능적 도메인으로 이루어져 있음을 알 수 있었다 또한 C-terminal의 10kD은 PDPc의 활성에는 영향을 주지 않는 것으로 밝혀졌으나 다른 도메인의 기능은 더 연구가 되어져야 할 것으로 생각된다.