Post Prandial Plasma Free Arginine Concentrations Increase in Rainbow Trout Fed Arginine-deficient Diets

Park, Gunjun,Bai, Sungchul C.,Ok, Im-ho,Han, Kyungmin,Hung, Silas S.O.,Rogers, Quinton R.,Min, Taesun Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.3

Three experiments were conducted to determine the effects of dietary arginine concentrations on plasma free amino acid (PAA) concentrations in rainbow trout, Oncorhynchus mykiss (Walbaum). The first experiment was conducted to determine appropriate post-prandial and food deprivation sampling times in dorsal aorta cannulated rainbow trout averaging 519${\pm}$9.5 g (mean${\pm}$SD) at $16^{\circ}C$. Blood samples were taken at 0, 2, 3, 4, 5, 6 and 24 h after feeding (0 and 24 h blood samples were taken from the same group of fish). PAA concentrations increased by 2 h post-feeding and the concentration of all essential amino acids except histidine peaked at 5 h and returned to 0 time values by 24 h. In the second experiment dorsal aorta cannulated rainbow trout averaging 528${\pm}$11.3 g (mean${\pm}$SD) were divided into 6 groups of 4 fish to study the effect of dietary arginine levels on PAA. After 24 h food deprivation, each group of fish was fed one of six L-amino acid diets containing graded levels of arginine (0.48, 1.08, 1.38, 1.68, 1.98 or 2.58%) by intubation. Blood samples were taken at 0, 5 and 24 h after feeding. Post-prandial (5 h after feeding) plasma-free arginine concentrations (PParg) showed a breakpoint at 1.03% arginine in the diet and post-absorptive (24 h after feeding) plasma free-arginine concentrations (PAarg) showed a breakpoint at 1.38% arginine. PAarg increased linearly from fish fed diets containing arginine between 0.48% and 1.38%, and the concentrations remained constant from fish fed diets containing arginine at or above 1.38%, but were all below PParg at all time points. Results of the third experiment confirm the results that PParg concentrations from fish fed arginine deficient diets were higher than PAarg (0 or 24 h values). Thus, in contrast to mammals and birds, the PParg when arginine is present in the diet as the most limiting amino acid such that it severely limits growth, increases in plasma rather than decreases.

성장기 암컷 쥐에서 Arginine 첨가가 골밀도에 미치는 영향

최미자(Choi Mi-Ja) 韓國營養學會 2007 Journal of Nutrition and Health Vol.40 No.3

The aim of this study was to define an arginine effect when added to a diet. The influence of arginine supplements on bone mineral density and content were studied in young female Sprague-Dawley rats fed either an arginine supplemented diet or control diet. Twenty four rats (body weight 83 ± 5 g) were randomly assigned to one of two groups, consuming casein or casein with supplemented arginine diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PlXImus (GE Lunar Co, Wisconsin, USA) in spine and femur 3, 6, and 9 weeks after feeding. The serum and urine concentrations of Ca and P were determined. Diet did not affect weight gain and mean food intake. The serum concentration of Ca and P were not changed by arginine supplementation. Urinary Ca excretion was significantly decreased by arginine supplementation. Spine BMO was significantly increased by arginine supplementation on 3 and 6 weeks after feeding. Femur BMD was significantly increased in the group of arginine supplementation on 3, 6, and 9 weeks. Rats fed the arginine-supplemented diet had better bone mineral content than did control diet rats in the experimental period. Therefore, arginine supplementation may be beneficial on spine and femur BMD increment in growing female rats. These are thought to be associated with an arginine-induced growth hormone release. The exact mechanism of this effect remains to be elucidated. (Korean J Nutr 2007; 40(3): 235~241)

난소절제쥐에서 Arginine 첨가 식이가 골밀도 및 골대사에 미치는 영향

최미자(Choi Mi-Ja) 韓國營養學會 2009 Journal of Nutrition and Health Vol.42 No.4

As far as we know, there were no studies of the effect of L-arginine on bone metabolism in post-menopausal women or ovariectomized rats. The primary objective of the current study was to determine whether arginine supplementation was associated with alterations in femoral and spinal bone mineral density (BMD) and bone markers in ovariectomized (Ovx) rats. Forty female Sprague-Dawley rats were divided into two groups, Ovx and sham groups, which were each randomly divided into two subgroups that were fed control and arginine supplemented diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. Bone resorption was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Serum osteocalcin, growth hormone, insulin-like growth factor-1 (IGF-1), parathyroid hormone (PTH) and calcitonin were analyzed using radioimmunoassay kits. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PIXImus (GE Lunar Co, Wisconsin, USA) in spine and femur. The serum and urine concentrations of Ca and P were determined. The plasma was analyzed for arginine. Diet did not affect weight gain, mean food intake, and plasma arginine concentration. Urinary Ca excretion was decreased by arginine supplementation in Ovx rats, but statistically not significant. The Ovx rats fed arginine-supplemented diet were not significantly different in ALP, osteocalcin, crosslinks value, PTH, calcitonin and IGF-1 compared to those fed control diet. The arginine-supplemented group had significantly higher serum Ca and growth hormone than control group. Spine and femur BMD were significantly increased by arginine supplementation on 5th and 9th weeks after feeding. Our findings indicate that dietary L-arginine supplementation decreased bone mineral density loss in Ovx rats. Therefore, dietary arginine supplementation may represent a potentially useful strategy for the management of osteoporosis.

Purification and Some Properties of Arginine Deiminase in Euglena gracilis Z

Park, Bong-Sun,Hirotani, Aiko,Nakano, Yoshihisa,Kitaoka, Shozaburo Korea Organic Resources Recycling Association 1993 유기물자원화 Vol.1 No.1

Euglena gracilis 에서 arginine deiminas는 mitochondrial matrix 내에 존재한다. 고도로 정제된 효소가 0.23 nM의 $K_m$ 값을 갖고 효소반응을 하기 위해서는 $Co^{2+}$가 필요하며, 이때 최적 pH는 9.3~10.3이었다. Gel filtration에 의해서 얻어진 조효소 단백질의 분자량은 87,000이었으며, SDS-acrylamide gel electrophoresis에 의해 효소는 48,000의 분자량을 갖는 2개의 동일한 subunit로 구성되어 있음이 밝혀졌다. Euglena의 arginine deiminas는 sulfhydryl inhibitors에 의해서 활성이 저지되었는데, 이는 sulfhydryl group이 효소의 활성부위에 관여함을 나타낸다. 이 sulfhydryl group은 arginine이 효소와 결합하는데 있어서 negative cooperativity를 나타내었다. ${\beta}-guanidinopropionate$, ${\gamma}-guanidinobutyrate$와 guanidinosuccinate는 효소의 활성을 저지시키지 않는데 반하여, $L-^{\alpha}-amino-{\beta}-guanidino-propionate$, D-arginine, 그리고 L-homoarginine은 효소의 활성을 강력하게 저지시켰다. Citrulline과 ornithine에 의해서도 상당한 정도의 효소활성저지가 관찰되었다. 우리는 Euglena의 arginine deiminase의 독특한 성질이 Euglena 라는 원생동물 내에서 arginine 대사의 조절에 어떻게 영향을 미치는지를 토의하고자 한다. In Euglena gracilis arginine deiminase was located in the mitochondrial matrix. The highly purified enzyme required $Co^{2+}$ for the enzyme reaction with the $K_m$ value of 0.23 nM, and its optimum pH was 9.7 to 10.3. The molecular weight of the native enzyme protein was 87,000 by gel filtration, and SDS-acrylamide gel electrophoresis showed that the enzyme consisted of two identical subunits with a molecular weight of 48,000. Euglena arginine deiminase was inhibited by sulfhydryl inhibitors, indicating that a sulfhydryl group is involved in the active center of the enzyme. It exhibited negative cooperativity in binding with arginine. $L-{\alpha}-amino-{\beta}-guanidino-propionate$, D-arginine, and L-homoarginine strongly inhibited the enzyme while ${\beta}-guanidinopro-pionate$, ${\gamma}-guanidinobutyrate$, and guanidinosuccinate did not. Considerable inhibition was also observed with citrulline and ornithine. We discuss the effects of the unique properties of the Euglena arginine deiminase on the regulation of arginine metabolism in this protozoon.

Euglena graci I is Z 로부터 Arginine Deiminase의 정제 및 그의 특성

Bong-Sun Park,Aiko Hirotani,Yoshihisa Nakano,Shozaburo Kitaoka 유기성자원학회 1993 유기물자원화 Vol.1 No.1

In Euglena gracilis arginine deiminase was located in the mitochondrial matrix. The highly purified enzyme required C02+ for the enzyme reaction with the Km value of 0.23mM, and its optimum pH was 9.7 to 10.3. The molecular weight of the native enzyme protein was 87,000 by gel filtration, and SDS-acry lamide gel electrophoresis showed that the enzyme consisted of two identical subunits with a molecular weight of 48 ,000. Euglena arginine deiminase was inhibited by sulfhydryl inhibitors, indicating that a sulfhydryl group is involved in the active center of the enzyme. It exhibited negative cooperativity in binding with arginine. L-a-amino-β, guanidino-propionate, D-arginine, and L-homoarginine strongly inhibited the enzyme while βguanidinopro-pionate , )tguanidinobutyrate, and guanidinosuccinate did not. Considerable inhibition was also observed with citrulline and ornithine. We discuss the effects of the unique properties of the Euglena arginine deiminase on the regulation of arginine metabolism in this protozoon. Euglena gracílis 에서 arginine deiminase는 mitochondrial matrix 내에 존재한다. 고도로 정제된 효소가 0.23 mM의 Km 값을 갖고 효소반응을 하기 위해서는 Co2 +가 필요하며, 이때 최적 pH는 9. 3~ 1O. 3이었다. Gel filtration에 의해서 얻어진 조효소 단백질의 분자량은 87, 000이었으며, SDSacry lamide gel electrophoresis에 의해 효소는 48, 000의 분자량을 갖는 2개의 동일한 subunit로 구성되어 있음이 밝혀졌다. Euglena의 arginine deiminase는 sulfhydryl inhibitors에 의해서 활성이 저지되었는데, 이는 sulfhydryl group이 효소의 활성부위에 관여함을 나타낸다. 이 sulfhydryl group은 arginine이 효소와 결합하는데 있어서 negative cooperativity를 나타내었다. ß-guanidinopropionate, Y-guanidinobutyrate와 guanidinosuccinate는 효소의 활성을 저지시키지 않는데 반하여, L-a -amino-ß-guanidino-propionate, D-arginine, 그리고 L-homoarginine은 효소의 활성을 강력하게 저지시켰다. Citrulline과 ornithine에 의해서도 상당한 정도의 효소활성저지가 관찰되었다. 우리는 Euglena의 arginine deiminase의 독특한 성질이 Euglena 라는 원생동물 내에서 arginine 대사의 조 절에 어떻게 영향을 미치는지를 토의하고자 한다.

알코올 대사 효소 alcohol dehydrogenase (ADH) 및 acetaldehyde dehydrogenase (ALDH) 활성에 미치는 아미노산의 영향

차재영(Jae-Young Cha),정해정(Hae-Jung Jung),정재준(Jae-Jun Jeong),양현주(Hyun-Ju Yang),김용택(Yong-Taek Kim),이용수(Yong-Soo Lee) 한국생명과학회 2009 생명과학회지 Vol.19 No.9

본 연구에서는 숙취해소에 좋은 것으로 알려진 식품 소재의 주요 아미노산을 포함하여 효소 활성에 영향을 미치는 것으로 알려진 아미노산을 선택하였고, 효소 활성도가 상대적으로 높은 yeast와 rat liver 유래의 ADH 및 ALDH 효소를 대상으로 알코올 대사에 관련된 효소 활성의 촉진 효과에 대하여 검토하였다. Rat liver 유래의 ADH 활성은 처리한 아미노산 중에서 arginine에서 가장 높았다. Arginine의 첨가 농도를 달리하여 효소 활성을 측정한 결과 10~50 ㎎/㎖ 농도에서 118~120.6%로 양성대조구의 90.6% 보다 약간 높은 것으로 나타났다. 또한, yeast 유래의 ADH 활성은 methionine에서 가장 높은 활성을 보였고, methionine의 처리 농도를 달리한 경우에서는 첨가 농도 의존적으로 높은 활성을 보였다. Rat liver 유래의 ALDH 활성은 methionine이 가장 높은 활성을 보였다. Methionine의 첨가 농도별 측정에서는 10 ㎎/㎖에서 30 및 50 ㎎/㎖ 첨가 농도에서 보다 높은 활성을 보였으며, 이들 모든 처리 농도에서 양성대조구 보다 상당히 높은 활성을 보였다. 한편 yeast 유래의 ALDH 활성은 각 아미노산별 큰 차이는 없었으나, arginine에서 높은 활성을 보였다. Arginine의 첨가 농도별 측정에서는 처리 농도 의존적으로 활성이 약간씩 증가하는 경향을 보였으며, 양성대조구 보다 높은 활성을 나타내었다. 효모 유래 ALDH 및 rat liver 유래 ADH 효소 활성을 촉진시키는 작용을 가진 arginine을 효모 배양에 첨가시킬 경우 세포 내 ALDH 및 ADH 활성 염색 정도가 증가함으로써 arginine은 ALDH 및 ADH 활성을 촉진시키는 효능이 in vivo 실험계에서도 확인되었다. 이상의 실험 결과에서 아미노산 중에서는 arginine과 methionine이 ADH 및 ALDH 활성을 촉진시키는 작용에 의해 알코올 분해뿐만 아니라 acetaldehyde의 분해도 촉진시킬 가능성이 높아 숙취해소 효과는 물론 간 보호 효과도 동시에 있을 것으로 시사 되어 진다. 따라서 arginine과 methionine과 같은 아미노산을 주류 제품에 첨가하게 될 경우 숙취해소 경감과 간 보호 효능을 어느 정도 나타낼 수 있을 가능성이 제기되었다. The present study examined the comparative effects of various amino acids on the alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities of yeast Saccharomyces cereviciae and rat liver homogenate in vitro. Methionine showed the highest activity in yeast ADH among the amino acids used in this study, but this was not higher than that of the hangover product, Condition-Power (CP) used as positive control. Methionine was also found to be the best amino acid in terms of the ALDH activity in rat liver homogenate among the treatment amino acids, which was comparatively higher than that of positive control CP. It was chosen for further experiments and yeast ADH activity increased in parallel with increased methionine concentration, but not rat liver ALDH activity, and it was comparatively higher than those of the positive control. Arginine showed the highest values in yeast ALDH and rat liver ADH activities among amino acids, and it was chosen for further experiments. Yeast ALDH activity increased in parallel with increased arginine concentration, which was higher than that of positive control CP, and rat liver ADH activity was also comparatively higher in all treatment concentrations of arginine than that of positive control CP. The native electrophoresis of ADH and ALDH from cell-free extracts of yeast Saccharomyces cerevisiae cultured in the growth medium containing various arginine concentrations by 0~0.1% showed two active bands upon zymogram staining analysis, and the straining intensity of ADH and ALDH active bands in arginine treatment yeast was stronger than that of non-yeast or low treatment yeast. These results indicate that alcohol metabolizing enzyme activities can be enhanced by arginine and methionine, suggesting that arginine and methionine have potent ethanol-metabolizing activities.

Arginine이 가토 심방근 수축력 및 동방결절세포의 전기생리학적 특성에 미치는 효과

김기순,강석한,고상돈,김진혁 한양대학교 의과대학 1991 한양의대 학술지 Vol.11 No.2

In other to explore effects of arginine, one of the basic amino acid on the cardiac function, changes in atrial contractility and action potentials of the sinus noed cells were determined in the isolated rabbit heart following superfusion with Tyrode's solution containing different amount of arginine. After superfusion with arginine (30mM) the amplitued of action potential (APA) of the sinus node cells generally decreased as a result of a greater decrement in maximum diastolic potential (MDP) than a simultaneous increment of avershoot potential (OS). No sigvificant changes were observed in action potential duration (APD) and rate of spontaneous firing (RSF) of the sinus node cells after superfusion with arginine solutions. At low concentration (3-6mM) arginine decreased atrial contractility while at high concentration (30mM) an initial positive inofropic response (between 3 and 5 min after onset of superfusion )was followed by a negative response. On the other hand, arginine intensified the initial positive inotropic response induced by reducing stimulation frequency of the atrial segment from 2Hz to 0.5Hz. And the negative inotropic effect of arginine (6mM) was abolished in the rabbir atrial segment pretreated with caffeine. The results of present study suggest that arginine can act diretly on the cardiac cells of rabbir heart resulting in an alteration lf transmenbrane potential and atrial contractility.

성장기 암컷 쥐에서 Arginine 첨가 식이가 골 대사 지표 및 호르몬에 미치는 영향

최미자(Choi Mi-Ja) 韓國營養學會 2007 Journal of Nutrition and Health Vol.40 No.4

An important related question is whether arginine has influence bone metabolism. The effect of arginine supplements on bone markers and related hormones were studied in young female Sprague-Dawley rats fed either an arginine supplemented diet or control diet. Twenty four rats (body weight 83±5 g) were randomly assigned to one of two groups, consuming casein or casein with supplemented arginine diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. And bone resorption rate was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Serum osteocalcin, growth hormone, estrogen, insulin-like growth factor-1 (1GF-1), parathyroid hormone (PTH) and calcitonin were analyzed using radioimmunoassay kits. The weight gain and mean food intake were not affected regardless of diets. The rats fed arginine-supplemented diet had not significantly different in ALP, osteocalcin, crosslinks value, PTH, estradiol, and IGF-1 compared to those fed casein diet group. The arginine-supplemented group had significantly higher growth hormone and calcitonin than casein group. This study suggests that arginine is beneficial for bone formation in growing female rats. Therefore exposure to diet which rich in arginine early in life may have benefits for bone formation and osteoporosis prevention. (Korean J Nutr 2007; 40(4): 320~326)

Targeting Arginine-Dependent Cancers with Arginine-Degrading Enzymes: Opportunities and Challenges

Melissa M. Phillips,Michael T. Sheaff,Peter W. Szlosarek 대한암학회 2013 Cancer Research and Treatment Vol.45 No.4

Arginine deprivation is a novel antimetabolite strategy for the treatment of argininedependent cancers that exploits differential expression and regulation of key urea cycle enzymes. Several studies have focused on inactivation of argininosuccinate synthetase 1 (ASS1) in a range of malignancies, including melanoma, hepatocellular carcinoma (HCC), mesothelial and urological cancers, sarcomas, and lymphomas. Epigenetic silencing has been identified as a key mechanism for loss of the tumor suppressor role of ASS1 leading to tumoral dependence on exogenous arginine. More recently, dysregulation of argininosuccinate lyase has been documented in a subset of arginine auxotrophic glioblastoma multiforme, HCC and in fumarate hydratasemutant renal cancers. Clinical trials of several arginine depletors are ongoing, including pegylated arginine deiminase (ADI-PEG20, Polaris Group) and bioengineered forms of human arginase. ADI-PEG20 is furthest along the path of clinical development from combinatorial phase 1 to phase 3 trials and is described in more detail. The challenge will be to identify tumors sensitive to drugs such as ADI-PEG20 and integrate these agents into multimodality drug regimens using imaging and tissue/fluid-based biomarkers as predictors of response. Lastly, resistance pathways to arginine deprivation require further study to optimize arginine-targeted therapies in the oncology clinic.

고콜레스테롤 식이 흰쥐의 과민성방광에서 Aspirin과 Arginine의 병용투여가 방광내압검사와 방광근수축에 미치는 영향

손환철,양지현,김수웅,백재승,구자현,박관진 대한배뇨장애요실금학회 2005 International Neurourology Journal Vol.9 No.1

Purpose: The purpose of this study was to show the effects of aspirin and arginine on hyperactive bladder of hypercholesterol diet rats. Materials and Methods: We used 40, 3-month-old Sprague-Dawley rats. Ten of them received the 2% cholesterol diet for 8 weeks and 10 received cholesterol diet and aspirin treatment and 10 received cholesterol diet, aspirin and arginine treatment. The remaining 10 served as control and were fed a normal diet. Cystometry and bladder muscle strips study were evaluated in each group. Results: Compared with normal control, mean serum cholesterol and body weight significantly elevated in the cholesterol group. Aspirin +/- arginine treatment lessened the increase of body weight and serum cholesterol level. Compared to control, the cholesterol group showed a shorter voiding interval and smaller functional bladder capacity in cystometrogram, however, aspirin +/- arginine treatment group showed no difference in voiding interval and functional bladder capacity. In detrusor muscle strip study, the increase in the proportion of purinergic components and the decrease in the proportion of cholinergic components were observed in the cholesterol group, however, not in aspirin +/- arginine treatment group. Overall, additional effect of arginine on aspirin treatment was negligible. Conclusion: Treatments of aspirin +/- arginine showed significant protective effect on the bladder overactivity induced by hypercholesterol diet in rats. In patients with heart diseases and overactive bladder, aspirin could be a useful treatment for protection and treatment. (J. Korean Continence Society 2005;9:6-12)