• Title/Summary/Keyword: ${\gamma}$-glutamyl kinase

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Isolation of a High-Yield Mutant Strain for L-Proline Production and Its Fermentation Conditions

  • Ryu, Wuk-Sang;Jang, Hyung-Wook;Cho, Kyoung-Hee;Chang, Soon-Jae;Ryu, Yeon-Woo;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.613-618
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    • 1999
  • L-Proline-producing mutant strains were developed by exposing L-glutamic acid-producing bacteria to N-metyl-N-nitro-nitrosoguanidine and UV irradiation. A L-histidine auxotroph of Corynebacterium acetoacidophilum RYU3161(KCTC 0616BP), which was resistant to sulfaguanidine and proline analogs (DHP, AZC, TAC), was isolated. The activity of the mutant strain's $\gamma$-glutamyl kinase was 45% higher than that of the parent strain. The optimum level of L-histidine for production of L-proline was 0.16 g/l. In a 5-1 jar fermenter, the mutant strain produced L-proline at a high concentration (35 g/l) level within 48 h of cultivation.

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In-vitro Production of Glutathione Using Yeast ATP Regeneration System and Recombinant Synthetic Enzymes from Escherichia coli. (효모의 ATP 재생산계와 대장균 유래의 재조합 생산효소를 이용한 in vitro 글루타치온 생산)

  • 고성영;구윤모
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.213-220
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    • 1998
  • An ATP regeneration system was used for the production of glutathione which was synthesized by a sequential action of ${\gamma}$-glutamyl-cysteine synthetase and glutathione synthetase. The synthetases above were produced in the recombinant E. coli (TG1/pDG7) with the highest specific production yield of 31 mg glutathione/g wet cell. Bakers yeast was considered to have economically a better ATP regeneration system although the glutathione production yield was lower than that of acetate kinase. It was also observed that the ATP regeneration system of bakers yeast was superior to that of Saccharomyces cerevisiae ATCC24858. The yield of glutathione production with bakers yeast was 36% with the ATP concentration of 5 mM. To avoid the cysteine limitation during the early phase of glutatione production, an extra cysteine was added at 2 hours after reaction and the production yield increased 1.91 times. The effectiveness of bakers yeast as an ATP regeneration system was proved by several sets of extra feeding experiments. The product inhibition by glutathione above 14 mM was also observed.

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Tissue level of chloramphenicol and haematological changes in olive flounder, Paralichthys olivaceus orally administered chloramphenicol (Chloramphenicol의 경구투여에 따른 양식 넙치, Paralichthys olivaceus의조직내 잔류량 및 혈액학적 변화)

  • Jeong, So-Jeong;Seo, Jeong-Su;Eom, Hye-Gyeong;Kim, Na-Yeong;Lee, Sang-Hwan;Heo, Min-Do;Jeong, Hyeon-Do;Jeong, Jun-Gi
    • Journal of fish pathology
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    • v.17 no.2
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    • pp.113-121
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    • 2004
  • This study was performed to investigate the pathphysiological side effects of chloramphenicol (CAP) on olive flounder, Paralichthys olivaceus. 150 mg/kg/bw/day of CAP was orally administered forcedly to flounder for 4 days and then the flounder was monitored for 11 days. The level of hematocrit was significantly decreased from 2 day to 3 day of CAP treatment in CAP-treated group. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and gamma glutamyl transferase (GGT) were slightly increased during the treatment of CAP and gradually decreased after the treatment of CAP. The levels of blood urea nitrogen (BUN) and creatinine kinase (CK), although not significant, were slightly increased during the treatment of CAP. The level of lactate dehydrogenase (LDH) was not changed during the treatment of CAP but slightly increased after the treatment of CAP. However, no histopathological changes were observed in various organ after the treatment. From these results, it is suggest that CAP can evoke several side effects on haematopoietic organs in olive flounder.

Effects of exercise on hematological and serum biochemical parameters in riding ponies (승용마의 운동에 따른 혈액학 및 혈청 생화학치의 변화)

  • Ko, Jeong-Ja;Lee, Young-Woo;Seo, Jong-Pil;Lee, Kyoung-Kap
    • Korean Journal of Veterinary Research
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    • v.60 no.1
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    • pp.33-37
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    • 2020
  • The purpose of the present study was to investigate changes in hematological and serum biochemical parameters in ponies exercising 3 times a day. The study included 10 healthy 4- to 5-year-old Jeju crossbred ponies used in riding lessons at an equestrian riding school. Hematology and serum chemistry samples were obtained before the initial lesson and after the last lesson of the day. The post-exercise results showed that packed cell volume, white blood cell, red blood cell, hemoglobin, and total protein levels increased significantly (p < 0.05). Serum Na+ also increased significantly (p < 0.01), but serum Ca++ decreased significantly (p < 0.01). Creatinine kinase, aspartate amino transferase, gamma glutamyl transferase, blood urea nitrogen, creatinine, and bilirubin levels increased significantly (p < 0.05), but the glucose level decreased significantly (p < 0.05). Furthermore, the serum cortisol hormone level increased significantly (p < 0.01). The results suggest that participating in riding lessons three times a day may result in various physiological changes, indicating the presence of exercise-related stress in riding ponies.

A Study on the Changes of Blood Mineral and Enzyme Activity During Growth Period of Velvet Antler in Korean Spotted Deer(Cervus Nippon) (꽃사슴의 녹용 성장기간 중 혈액 내 미네랄 및 효소활성 변화에 관한 연구)

  • Kim, M.H.;Moon, S.H.;Lee, C.H.;Jeon, B.T.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.1031-1038
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    • 2003
  • This study was aimed at investigating the change of blood mineral and enzyme activity during growth period of velvet antler in Korean spotted deer. Samples of blood, obtained from the jugular vein of twenty-five deer(4 to 6 year-old males), were taken in 10 days interval from just after casting to 50 days. Deer were randomly selected from the farm, and samples were analyzed for blood parameters like mineral concentration and enzyme activities. No significant differences found in calcium and phosphorus concentration in blood whereas sodium, potassium and chloride concentration were significantly changed with antler growth. There were no significant differences in alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, $\gamma$-glutamyl transferase and lactate dehydrogenase during growth of antler, but alkaline phosphatase concentration was increased with growth of antler, and the highest concentration was obtained on the 50 days after casting. Creatine kinase and lactate dehydrogenase activities for the deer tested in this experiment were higher than those of other animals.

Correlation between Arterial Stiffness and Physiological Parameters (동맥경화도와 생리학적 변수들 간의 상관관계)

  • Shin, Jae-Wook;Seok, Seong-Ja;Lee, Gil-Hyun;Choi, Seok-Cheol;Hyun, Kyung-Yae
    • The Korean Journal of Health Service Management
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    • v.7 no.3
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    • pp.71-82
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    • 2013
  • Arterial stiffness(AS) is an important pathologic state of vascular injury. This study was carried out to elucidate the effect of physiological variables on brachial-ankle pulse wave velocity(BAPWV), index of AS. Four hundred adults(volunteers) participated in this study. Body indices, biochemical, cardiac and inflammatory markers, and right(Rt)- and left(Lt)-BAPWV were measured. Body mass index(BMI), Rt- and Lt-BAPWV, glucose, triglyceride, alkaline phosphatase(ALP), gamma-glutamyl transferase(GGT), creatinine, uric acid, troponin-I(TNI), NT-proBNP and high sensitivity C-reactive protein(hs-CRP) levels were higher than the reference value of each variable. Rt- and Lt-BAPWV were directly correlated with age, body weight, BMI, glucose, ketone, aspartate aminotransferase, alanine aminotransferase, ALP, GGT, total cholesterol, low density lipoprotein, lipoprotein(a), apolipoprotein-B, blood urea nitrogen, heart rate, TNI, creatine kinase, CK-MB, lactic dehydrogenase, myoglobin, hs-CRP, lipase, reumatoid factor, fibrinogen and D-dimer (P<0.05, P<0.01, P<0.001 or P<0.000, respectively), but inversely associated with total bilirubin, uric acid, apolipoprotein-A1 and GFR (P<0.05). These observations suggest that a variety of physiological variables may influence BAPWV, resulting in increased risk or prevention of cardiovascular and/or cerebrovascular attacks. Therefore, physiological variables affecting BAPWV should be regularly controlled.

Biochemical, Haematological and Thyroid Hormone Profile in Healthy Indian Kathiawari Horses

  • Gupta, A.K.;Kumar, Sanjay;Pal, Yash
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.8
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    • pp.1215-1221
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    • 2002
  • Normal haematological and biochemical indices along with thyroid hormone status were studied in healthy Kathiawari horses of different age groups (yearling, young stock, adults and old stock) belonging to either sex. Effect of both age and sex was observed on thyroid hormone levels, haematological and biochemical indices. In females, hemoglobin levels was significantly lower in yearlings than adult animals while total leukocyte counts were higher in yearlings than equids of other age groups. Sex had effect only on total erythrocyte counts, mean corpuscular hemoglobin concentration and mean corpuscular hemoglobin in horses of 1-3 years age group (young stock) and on packed cell volume in adult female and male equids. Among biochemical indices, activities of enzymes were observed to be influenced both by age and sex. Creatine kinase, gamma glutamyl transferase, glutamate pyruvate transaminase, glutamic oxaloacetate transaminase and lactate dehydrogenase activities were significantly higher in young and adult equids than animals of other age groups in Kathiawari horses while activity of alkaline phosphatase was significantly higher in yearlings than equids belonging to other age groups in both male and females. However, activity of sorbitol dehydrogenase was unaltered due to both sex and age factor. Albumin, bilirubin direct, bilirubin total, cholesterol, creatinine, protein, triglyceride and uric acid were statistically different in various age and sex groups of horses. Calcium, magnesium and chloride contents were almost same in various age groups of male horses. Significantly higher levels of $T_3$ and $T_4$ were observed in both male and female yearlings as compared to equids of other age groups in both the sexes.

Regulatory Mechanisms of Angiotensin II on the $Na^+/H^+$ Antiport System in Rabbit Renal Proximal Tubule Cells. I. Stimulatory Effects of ANG II on $Na^+$ Uptake

  • Han, Ho-Jae;Koh, Hyun-Ju;Park, Soo-Hyun
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.413-423
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    • 1997
  • The importance of the kidney in the development of hypertension was first demonstrated by Goldblatt and his colleagues more than fifty years ago. Many hormones and other regulatory factors have been proposed to play a major role in the development of hypertension. Among these factors angiotensia II (ANG II) is closely involved in renal hypertension development since it directly regulates $Na^+$ reabsorption in the renal proximal tubule. Thus the aim of the present study was to examine signaling pathways of low dose of ANC II on the $Na^+$ uptake of primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined seum-free medium. The results were as follows: 1) $10^{-11}$ M ANG II has a significant stimulatory effect on growth as compared with control. Alkaline phosphatase exhibited significantly increased activity. However, leucine aminopeptidase and ${\gamma}-glutamyl$ transpeptidase activity were not significant as compared with control. In contrast to $10^{-11}$ M ANG II stimulated $Na^+$ uptake $(108.03{\pm}2.16% of that of control)$, $10^{-9}$ M ANG II inhibited ($92.42{\mu}2.23%$ of that of control). The stimulatory effect of ANG II on $Na^+$ uptake was amiloride-sensitive and inhibited by losartan (ANG II receptor subtype 1 antagonist) and not by PD123319 (ANG II receptor subtype 2 antagonist). 2) Pertussis toxin (PTX) alone inhibited $Na^+$ uptake by $85.52{\pm}3.52%$ of that of control. In addition, PTX pretreatment prevented the AMG II-induced stimulation of $Na^+$ uptake. 8-Bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP), forskolin, and isobutylmethylxanthine (IBMX) alone inhibited $Na^+$ uptake by $88.79{\pm}2.56,\;80.63{\pm}4.38,\;and\;84.47{\pm}4.74%$ of that of control, respectively, and prevented the ANG II-induced stimulation of $Na^+$ uptake. However, $10^{-11}$ M ANG II did not stimulate cAMP production. 3) The addition of 12-O-te-tradecanoylphorbol-13-acetate (TPA, 0.01 ng/ml) to the PTCs produced significant increase in $Na^+$ uptake ($114.43{\pm}4.05%$ of that of control). When ANG II and TPA were added together to the PTCs, there was no additive effect on $Na^+$ uptake. Staurosporine alone had no effect on $Na^+$ uptake, but led to a complete inhibition of ANG II- or TPA-induced stimulation of Na'uptake. ANG II treatment resulted in a $111.83{\mu}4.51%$ increase in total protein kinase C (PKC) activity. In conclusion, the PTX-sensitive PKC pathway is the main signaling cascade involved in the stimulatory effects of ANG II on $Na^+$ uptake in the PTCs.

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An Empirical Study of the Clinically Reportable Range in Clinical Chemistry (임상보고 가능범위의 실증적 연구)

  • Chang, Sang-Wu;Lee, Sang-Gon;Choi, Ho-Seong;Song, Eun-Young;Park, Yong-Won;Lee, In-Ae
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.1
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    • pp.31-36
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    • 2007
  • The purpose of the clinically reportable range (CRR) in clinical chemistry is to estimate linearity in working range. The reportable range includes all results that may be reliably reported, and embraces two types of ranges: the analytical measurement range (AMR) is the range of analyte values that a method can directly measure on the specimen without any dilution, concentration, or other pretreatment not part of the usual assay process. CAP and JCAHO require linearity on analyzers every six months. The clinically reportable range is the range of analyte values that a method can measure, allowing for specimen dilution, concentration, or other pretreatment used to extend the direct analytical measurement range. The AMR cannot exceed the manufacturer's limits. Establishing AMR is easily accomplished with Calibration Verification Assessment and experimental Linearity. For example: The manufacturer states that the limits of the AST on their instrument are 0-1100. The lowest level that could be verified is 2. The upper level is 1241. The verified AMR of the instrument is 2-1241. The lower limit of the range is 2, because that is the lowest level that could be verified by the laboratory. The laboratory could not use the manufacturer's lower limit of 2 because they have not proven that the instrument values below 2 are valid. The upper limit of the range is 1241, because although the lab has shown that the instrument is linear to 1241, the manufacturer does not make that claim. The laboratory needs to demonstrate the accuracy and precision of the analyzer, as well the validation of the patient AMR. Linearity requirements have been eliminated from the CLIA regulations and from the CAP inspection criteria, however, many inspectors continue to feel that linearity studies are a part of good lab practice and should be encouraged. If a lab chooses to continue linearity studies, these studies must fully comply with the calibration/calibration verification requirements of CLIA and/or CAP. The results of lower limit and upper limit of clinically reportable range were total protein (2.1 - 79.9), albumin (1.3 - 39), total bilirubin (0.2 - 106.2), alkaline phosphatase (13 - 6928.2), aspartate aminotransferase (24 - 7446), alanine aminotransferase (13 - 6724.2), gamma glutamyl transpeptidase (16.64 - 9904.2), creatine kinase (15.26 - 4723.8), lactate dehydrogenase (127.66 - 13231.8), creatinine (0.4 - 129.6), blood urea nitrogen (8.67 - 925.8), uric acid (1.6 - 151.2), total cholesterol (48.52 - 3162), triglycerides (36.91 - 3367.8), glucose (31 - 4218), amylase (21 - 6694.2), calcium (3.1 - 118.2), inorganic phosphorus (1.11 - 108), HDL (11.74 - 666), NA (58.3 - 1800), K (1.0 - 69.6), CL (38 - 1230).

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An Empirical Study of the Recovery Experiment in Clinical Chemistry (임상화학검사실에서 회수율 실험의 실증적 연구)

  • Chang, Sang-Wu;Lee, Sang-Gon;Song, Eun-Young;Park, Yong-Won;Park, Byong-Ok
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.184-188
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    • 2006
  • The purpose of the recovery experiment in clinical chemistry is performed to estimate proportional systematic error. We must know all measurements have some error margin in measuring analytical performance. Proportional systematic error is the type of error whose magnitude increases as the concentration of analyte increases. This error is often caused by a substance in the sample matrix that reacts with the sought for analyte and therefore competes with the analytical reagent. Recovery experiments, therefore, are used rather selectively and do not have a high priority when another analytical method is available for comparison purposes. They may still be useful to help understand the nature of any bias revealed in the comparison of kit experiments. Recovery should be expressed as a percentage because the experimental objective is to estimate proportional systematic error, which is a percentage type of error. Good recovery is 100.0%. The difference between 100 and the observed recovery(in percent) is the proportional systematic error. We calculated the amount of analyte added by multiplying the concentration of the analyte added solution by the dilution factor(mL standard)/(mL standard + mL specimen) and took the difference between the sample with addition and the sample with dilution. When making judgments on method performance, the observed that the errors should be compared to the defined allowable error. The average recovery needs to be converted to proportional error(100%/Recovery) and then compared to an analytical quality requirement expressed in percent. The results of recovery experiments were total protein(101.4%), albumin(97.4%), total bilirubin(104%), alkaline phosphatase(89.1%), aspartate aminotransferase(102.8), alanine aminotransferase(103.2), gamma glutamyl transpeptidase(97.6%), creatine kinase(105.4%), lactate dehydrogenase(95.9%), creatinine(103.1%), blood urea nitrogen(102.9%), uric acid(106.4%), total cholesterol(108.5), triglycerides(89.6%), glucose(93%), amylase(109.8), calcium(102.8), inorganic phosphorus(106.3%). We then compared the observed error to the amount of error allowable for the test. There were no items beyond the CLIA criterion for acceptable performance.

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