• Title/Summary/Keyword: Glycosylated

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Secretory Expression and Purification of the Recombinant Duck Interleukin-2 in Pichia pastoris

  • Du, Cuihong;Han, Long;Xiao, Anfeng;Cao, Minjie
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1264-1269
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    • 2011
  • Interleukin-2 (IL-2) is a vital cytokine secreted by activated T lymphocytes, and plays an important role in the regulation of cellular functions and immunity of animals. In this study, the recombinant duck IL-2 (rduIL-2) was secretory expressed in Pichia pastoris (P. pastoris). The recombinant P. pastoris strain was cultured in shake flasks and then scaled up in a 5.0-l bioreactor. The result showed that the maximal fresh-cell-weight of 594.1 g/l and the maximal $OD_{600}$ of 408 were achieved in the bioreactor. The rduIL-2 was purified by two steps of purification procedures, and approximately 311 mg of rduIL-2/L fermentation supernatant was obtained. SDS-PAGE showed that the purified rduIL-2 constituted a homogeneous band of ~16 kDa or ~14 kDa corresponding to the glycosylated or non-glycosylated duIL-2 protein in size, respectively. The bioactivity of rduIL-2 was determined by lymphocyte proliferation assay. The result indicated that the rduIL-2 greatly promoted the proliferation of ConA-stimulated lymphocytes in vitro. The P. pastoris expression system described here could provide promising, inexpensive, and large-scale production of the rduIL-2, which lays the foundation for development of novel immunoadjuvants to enhance both the immunity of ducks against various infectious pathogens and vaccine efficacy.

Exploring the Nucleophilic N- and S-Glycosylation Capacity of Bacillus licheniformis YjiC Enzyme

  • Bashyal, Puspalata;Thapa, Samir Bahadur;Kim, Tae-Su;Pandey, Ramesh Prasad;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1092-1096
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    • 2020
  • YjiC, a glycosyltransferase from Bacillus licheniformis, is a well-known versatile enzyme for glycosylation of diverse substrates. Although a number of O-glycosylated products have been produced using YjiC, no report has been updated for nucleophilic N-, S-, and C- glycosylation. Here, we report the additional functional capacity of YjiC for nucleophilic N- and S- glycosylation using a broad substrate spectrum including UDP-α-D-glucose, UDP-N-acetyl glucosamine, UDP-N-acetylgalactosamine, UDP-α-D-glucuronic acid, TDP-α-L-rhamnose, TDP-α-D-viosamine, and GDP-α-L-fucose as donor and various amine and thiol groups containing natural products as acceptor substrates. The results revealed YjiC as a promiscuous enzyme for conjugating diverse sugars at amine and thiol functional groups of small molecules applicable for generating glycofunctionalized chemical diversity libraries. The glycosylated products were analyzed using HPLC and LC/MS and compared with previous reports.

Effect of Trichloroacetic Acid on the Solubility of Caseinomacropeptide

  • Shin, Sung-Chul;Jang, Hae-Dong
    • Preventive Nutrition and Food Science
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    • v.7 no.1
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    • pp.43-47
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    • 2002
  • Crude caseinomacropeptide (CMP) was prepared from Na-caseinate using a commercial renneting enzyme. Most of the crude CMP was released from the Na-caseinate by hydrolyzing with the enzyme for 40 min. The hydrolysis of the k-casein with carbohydrate was slower than that of the k-casein without carbohydrate, as shown by the analyses of the sialic acid content and the tricine-SDS-polyacrylamide gel electrophoresis. The yield of crude CMP from Na-caseinate was 3.7%. Cation exchange chromatography showed that the crude CMP consisted of 40.5% CMP and 59.5% caseinogylcomacropetide (CGP). The effect of the TCA concentration on the solubility of CMP and CGP was determined by using crude CMP. The amounts of crude CMP and sialic acid decreased in the proportion to the increase of trichloroacetic acid (TCA) concentration from 2 to 12%, suggesting that the CGP containing carbohydrate, as well as the CMP having no carbohydrate, was precipitated in a range of 4 to 12%, depending on the TCA concentration. This result supports the hypothesis that the different non-glycosylated and glycosylated forms of CMP have different sensitivities to TCA precipitation.

Investigation of Angiotensin Glycosylation by MALDI-TOF and ESI Tandem Mass Spectrometry

  • Park, Soo-Jin;Park, Deok-Hie;Sul, Soo-Hwan;Oh, Sung-Hwan F.;Park, In-Sook;Chung, Doo-Soo;Kim, Hie-Joon;Kim, Min-Sik;Lee, Sang-Won
    • Bulletin of the Korean Chemical Society
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    • v.25 no.12
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    • pp.1791-1800
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    • 2004
  • Angiotensin I, a model decapeptide, was glycosylated and partially hydrolyzed with HCl (6 N, 80 $^{\circ}C$, 4 h), aminopeptidase, and carboxypeptidase Y. A single peptide mass map obtained from truncated peptides in the partial acid hydrolysate of angiotensin and its glycosylation product mixture by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry enabled sequencing of angiotensin by a combinatorial procedure. MALDI-TOF and electrospray ionization (ESI) tandem mass spectrometric results indicate that both the N-terminal amino group of aspartic acid and the guanidinium group of the second residue arginine are glycosylated.

A Survey of Knowledge of Diabetes in Patients with Type 2 Diabetes (제2형 당뇨병 환자의 질병관련 지식)

  • Kim Hee-Seung
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.11 no.1
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    • pp.67-73
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    • 2004
  • Purpose: The purpose of this study was to investigate the knowledge of diabetes and insulin use in patients with type 2 diabetes. Method: Participants (139) were recruited from the endocrinology outpatient department of a tertiary care hospital in an urban city. Knowledge of diabetes knowledge was measured by a brief diabetes knowledge test which had two components: a 14-item general test and a 9-item insulin-use subscale. Result: The average scores for knowledge of diabetes and insulin were 62.4 and 41.8 respectively. Scores were lower for items on the definition of glycosylated hemoglobin, diabetic diet, signs of ketoacidosis, and insulin reaction compared to other items. Knowledge scores were significantly lower for older participants, those who had lower levels of education, those who did not have a spouse, those who did not have a job, those who were not being treated with insulin, and those who tested their blood glucose less than 3 times a day. Conclusion: Definition of glycosylated hemoglobin, diabetic diet, signs of ketoacidosis, and insulin reaction should be included in education for patients with type 2 diabetes. Special education programs should be developed for elderly people, those with low levels of education those without a spouse, or without a job, or who are not being treated with insulin.

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Effect of Telephone Follow-up on Blood Glucose Improvement of Diabetes Mellitus Patients with Depressive Symptoms (전화추후관리가 우울성향을 가진 당뇨병 환자의 혈당개선에 미치는 효과)

  • Song, Min-Sun;Kim, Hee-Seung;Lee, Dong-Han
    • Journal of Korean Biological Nursing Science
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    • v.8 no.2
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    • pp.29-39
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    • 2006
  • The aim of this study was to examine the effect of telephone follow-up on glycemic control, lipid profiles. and treatment method in diabetic patients with depressive symptoms over one year follow-up period. Twenty-two diabetic patients attended the structured diabetes education program between March 2004 and February 2005 and completed a questionnaire using the Beck Depression Inventory(BDI) to measure depressive symptoms. The patients showed depressive symptoms with the criterion being a BDI score at least 16. The intervention was applied to the telephone follow-up for one year. Data were analyzed using Wilcoxon signed rank test and McNemar test. The patients decreased their mean glycosylated hemoglobin(HbAlc) levels by 1.9%(p=0.001). There was a significant mean change in the fasting blood sugar(FBS) level, with a mean change of -61mg/$d{\ell}$(p=0.008). These findings indicated that the telephone follow-up may be effective in the HbAlc and FBS control.

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Factors Associated with Lower Urinary Tract Symptoms in Patients with Type 2 Diabetes Mellitus (제2형 당뇨병 환자의 하부요로증상 관련요인)

  • Song, Hyo Jeong
    • Journal of Korean Biological Nursing Science
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    • v.17 no.1
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    • pp.71-77
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    • 2015
  • Purpose: This study was performed to identify lower urinary tract symptoms (LUTS), and to evaluate the factors affecting LUTS in patients with type 2 diabetes mellitus (T2DM). Methods: The cross sectional study was used with a structured questionnaire to collect data through interviews with 181 T2DM patients and their clinical data from a university hospital diabetes clinic from October 2010 to April 2012. LUTS were measured using the International Prostate Symptom Score (IPSS), depression using the Center for Epidemiologic Studies Depression Scale (CES-D), and glycosylated hemoglobin (HbA1c) from the clinical data. Results: Of all patients with T2DM, the mean IPSS of LUTS was $9.34{\pm}6.86$. Concerning the reported severity of LUTS, 53.6% of the subjects were in the moderate and severe group. In each symptom score of LUTS (range 0-5), nocturia was the highest 2.04, weak stream 1.62, and frequency 1.45. LUTS was significantly predicted by HbA1c and depression, and 14.3% of the variance in LUTS was explained. Conclusion: HbA1c and depression were found to be very important factors associated with LUTS in T2DM patients.

Studies on the Primary Structure of the Alkaline Protease in Neungee [Sarcodon aspratus (Berk.) S. Ito] I. Amino Acid Composition, Chemical Modification and Sequence of the N-terminal Amino Acid (능이[Sarcodon aspratus(Berk.) S. Ito]중 알카리성 단백질가수분해효소의 1차구조에 관한 연구 I. 아미노산 조성, 활성부위 아미노산 및 N-말단 부위의 아미노산 배열)

  • 이태규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.811-814
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    • 1993
  • Properties of a protease purified from Sarcodon asparatus(Berk.) S. Ito have been investigated. The enzyme displays as a glycosylated serine protease. The sequence for the 21 amino acids of the N-terminal side in the enzyme was determined by automated sequence analysis. The sequence was V-T-T-K-Q-T-N-A-P-W-G-L-G-N-I-S-T-T-N-K-L-.

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The Effect of Diabetes, Gestational Diabetes or Pre-eclampsia on Urinary Protein and Mineral Excretion during Pregnancy

  • Joo, Eun-Jung;Janet C. King
    • Preventive Nutrition and Food Science
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    • v.2 no.3
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    • pp.225-231
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    • 1997
  • Thirteen healthy control, 13 pre-eclamptic, 7 diabetic(DM) and 12 gestational diabetic(GDM) pregnant women participated in a study ofthe interrelationships between the levels of protein, calcium, magnesium, phosphorus, zinc and copper in urine. Urinary protein, magnesium and copper levels were significantly higher (p<0.0005, p<0.0003, p<0.005 respectively) in pre-eclamptic women than those of control, DM and GDM women. Urinary zinc excretion in pre-eclamptic women (1.61 mg/g creatinine) was higher than that of DM women (0.81mg/g creatinine); urinary zinc losses of control and GDM women were wre between the other two rups. The GDM women excreted significantly ore phosphorus in their urine in comparison to control and preeclamptic women (p<0.02), but this was not seen in DM women. Among the DM women, urinary protein excretion was positively correlated with glycosylated hemoglobin(r=0.940) and fasting blood glucose concentration (r=0.889). Urinary zinc excretion also was correlated with glycosylated hemoglobin (r=0.853) and fasting blood glucose (r=0.956). In the GDM and pre-eclamptic women there were also significant correlations between urinar calcium and magnesium (r=0.857, r=0.749 respectively) and between urinary protein and copper(r=0.638, r=0.778 respectively).

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Analysis of the relationship between lifestyle habits and glycosylated hemoglobin control based on data from a Health Management Plan

  • Wang, Ya-Chun;Wang, Chi;Shih, Ping-Wen;Tang, Pei-Ling
    • Nutrition Research and Practice
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    • v.14 no.3
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    • pp.218-229
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    • 2020
  • BACKGROUND/OBJECTIVES: Type 2 Diabetes mellitus (T2DM) is a hereditary disease that is also strongly dependent on environmental factors, lifestyles, and dietary habits. This study explored the relationship between lifestyle habits and glycosylated hemoglobin management in T2DM patients to provide empirical outcomes to improve T2DM management and patient health literacy. SUBJECTS/METHODS: This study enrolled 349 diabetic patients with more than 5 care visits to a Diabetes Mellitus care network under the Health Management Plan led by Taiwan Department of Health (DOH). Based on relevant literature, an Outpatient Record Form of Diabetes Mellitus Care was designed and lipid profile tests were conducted for data collection and analysis. RESULTS: When modeling the data, the results showed that the odds for HbA1c > 7.5% in T2DM patients duration over 10 years was 3.785 (P = 0.002) times that in patients with disease duration of fewer than 3 years. The odds of HbA1c > 7.5% in illiterate patients was 3.128 (P = 0.039) times that in patients with senior high school education or above. The odds of HbA1c > 7.5% in patients with other chronic illness was 2.207 (P = 0.019) times that in participants without chronic illness. Among 5 beneficial lifestyle habits, the odds of HbA1c > 7.5% in patients with 2 or 3 good habits were 3.243 (P = 0.003) and 3.424 (P = 0.001) times that in patients with more than 3 good habits, respectively. CONCLUSION: This empirical outcome shows that maintaining a good lifestyle improves T2DM management and patients' knowledge, motivation, and ability to use health information. Patients with longer disease duration, education, or good lifestyle habits had optimal HbA1c management than those in patients who did not. Thus, effective selfmanagement and precaution in daily life and improved health literacy of diabetic patients are necessary to increase the quality of T2DM care.