• Title/Summary/Keyword: Serine

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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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Partial Purification and Characterization of the Alkaline Protease from Baccillus sp. (Bacillus sp.가 생산하는 호알카리성 Protease의 부분정제 및 특성)

  • 안장우;오태광;박용하;박관하
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.344-351
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    • 1990
  • An alkalophilic microoganism producing a detergent-resistant alkaline protease was isolated from soil and identified as Baeiltus sp. The alkaline protease has been partially purified by ammonium sulfate fractionation, DEAE-Cellulose, CM-Cellulose and Sephdex G-100 column chromatography. The purified alkaline protease was highly active at pH 12-13 toward casein and stable at pH values from 6 to ll. The optimum temperature for the enzyme reaction was $55^{\circ}C$. The enzyme was completely inactivated by diisopropyl fluorophosphate (DFP) indicating that the enzyme was serine protease, but considerabiy stable in the presence of surface active agents.

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Expression and Purification of Recombinant Active Prostate-Specific Antigen from Escherichia coli

  • Jeong, Su-Jin;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.840-846
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    • 2007
  • Human prostate-specific antigen(PSA), a 33 kDa serine protease with comprehensive homology to glandular kallikrein, is secreted from prostatic tissue into the seminal fluid and enters into the circulation. The level of PSA increases in the serum of patients with prostatic cancer and hence is widely employed as a marker of the disease status. In particular, an enzymatically active PSA that is a form cleaved at the N-terminal seven-amino-acids prosequence, APLILSR, of proPSA may play an important roll in the progression of prostate cancer. Thus, the presence of the active form would selectively discriminate the cancer from benign prostatic hyperplasia. In this study, we developed a convenient purification method for the acquisition of active PSA and proPSA. Recombinant proPSA and active PSA were expressed directly in Escherichia coli, easily and efficiently isolated from inclusion bodies, refolded, and purified. Moreover, the enzymatic activity of the recombinant active PSA was confirmed as serine protease using chromogenic chymotrypsin substrate. This purified active PSA could be further applied to scrutinize the biological or conformational characteristics of the protein and to develop specific diagnostic and/or therapeutic agents against prostate cancer.

Altered Sulfate Metabolism of Arabidopsis Caused by Beet Severe Curly Top Virus Infection

  • Lee, Hong-Gun;Park, Sung-Hee;Kim, Dong-Giun;Lee, Taek-Kyun;Yum, Seung-Shic;Auh, Chung-Kyoon;Lee, Suk-Chan
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.355-360
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    • 2005
  • Sulfur, an important component of plants, is regulated by a variety of stresses in sulfate assimilation and metabolism. Increase has been observed in the expression of O-acetylserine(thiol)lyase (OASTL) through two-dimensional electrophoresis with the shoot tips of Arabidopsis infected by beet severe curly top geminivirus (BSCTV). With the three- to six-fold increases in the transcript expression of OASTL, serine acetyltransferase (SAT) and $\gamma$-glutmylcysteine synthetase (GSH) were induced over the mock-inoculated organization in each organization through real-time RT-PCR analysis. The expression of those genes might affect the accumulation of anthocyanin in symptomatic tissues and the induction of abnormal callus-like structures formed by additional cell divisions as typical disease symptoms of BSCTV-infected Arabidopsis. This is the first report to describe the collaborative induction of OASTL, SAT, and GSH in virus-infected plants. The changed expressions of OASTL, SAT, and GSH in Arabidopsis infected with BSCTV raises new aspects regarding the biological function of symptomatic tissues related to sulfate metabolism.

Changes of Proteolytic Enzyme Property by Gamma Irradiation (감마선 조사에 의한 단백질분해효소의 특성변화)

  • Yook, Hong-Sun;Lee, Hyun-Ja;Im, Sung-Il;Kim, Sung;Byun, Myung-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1116-1121
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    • 1997
  • Effects of gamma irradiation onf the activity and the properties(amino acid compositions, in vitro digestibility and SDS-PAGE pattern) of proteolytic enzymes were investigated. The proteolytic activity of soluble human serine protease, enzyme in kiwi and pineapple decreased 10% and 30~65% by 5 kGy and 30 kGy, respectively. In dried pancreatin and lysozyme, the proteolytic and antimicrobial activities decreased 6~14% and 10~20% by 5kGy and 40kGy, respectively. The analysis of above 10kGy-irradiated soluble human serine protease by SDS-PAGE revealed radiolysis of the enzyme into protein or peptides of lower molecular weights. The irradiation of skim milk, hammastein casein, and lysozyme up to 40kGy had no deleterious effect on either the in vitro digestibility or amino acid compositions.

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Functional Studies of Cysteine Residues in Human Glutathione S-Transferase P1-1 by Site-Directed Mutagenesis

  • Park, Hui Jung;Lee, Gwang Su;Gong, Gwang Hun
    • Bulletin of the Korean Chemical Society
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    • v.22 no.1
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    • pp.77-83
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    • 2001
  • To gain further insight into the relationship between structure and function of glutathione S-transferase (GST), the four cysteine mutants, C14S, C47S, C101S and C169S, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized glutathione (GSH). The catalytic activities of the four mutant enzymes were characterized with five different substrates as well as by their binding to four different inhibitors. Cys14 seems to participate in the catalytic reaction of GST by stabilizing the conformation of the active-site loop, not in the GSH binding directly. The substitution of Cys47 with serine significantly reduces the affinity of GSH binding, although it does not prevent GSH binding. On the other hand, the substitution of Cys101 with serine appears to change the binding affinity of electrophilic substrate by inducing a conformational change of the $\alpha-helix$ D. Cys169 seems to be important for maintaining the stable conformation of the enzyme. In addition, all four cysteine residues are not needed for the steroid isomerase activity of human glutathione S-transferase P1-1.

Primary metabolic responses in the leaves and roots of bell pepper plants subjected to microelements-deficient conditions

  • Sung, Jwakyung;Lee, Yejin;Lee, Seulbi
    • Korean Journal of Agricultural Science
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    • v.48 no.1
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    • pp.179-189
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    • 2021
  • Plants need essential mineral elements to favorably develop and to complete their life cycle. Despite the irreplaceable roles of microelements, they are often ignored due to the relative importance of macroelements with their influence on crop growth and development. We focused on the changes in primary metabolites in the leaves and roots of bell pepper plants under 6 microelements-deficient conditions: Copper (Cu), Zinc (Zn), Iron (Fe), Manganese (Mn), Boron (B) and Molybdenum (Mo). Bell pepper plants were grown in hydroponic containers, and individual elements were adjusted to 1/10-strength of Hoagland nutrient solution. A remarkable perturbation in the abundance of the primary metabolites was observed for the Fe and B and the Mn and B deficiencies in the leaves and roots, respectively. The metabolites with up-accumulation in the Fe-deficient leaves were glucose, fructose, xylose, glutamine, asparagine and serine. In contrast, the Mn deficiency also resulted in a higher accumulation of glucose, fructose, xylose, galactose, serine, glycine, β-alanine, alanine and valine in the roots. The B deficiency noticeably accumulated alanine, valine and phenylalanine in the roots while it showed a substantial decrease in glucose, fructose and xylose. These results show that the primary metabolism could be seriously disturbed due to a microelement deficiency, and the alteration may be either the specific or adaptive responses of bell pepper plants.

A Study on Salt-fermented Seahorse added with Proteolytic Enzyme (Protamex)

  • LEE, In-Sook;LEE, Min-Ho;JANG, Kyung-Tae
    • The Korean Journal of Food & Health Convergence
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    • v.6 no.6
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    • pp.1-7
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    • 2020
  • We compared the fermentation of 0 to 4 weeks by manufacturing a rapid low salt-fermented seahorse with a commercial Protamex added to the functional food, Hippocampus abdominalis. We studied amino acid composition, content and major amino acids related to flavor during the fermentation process of salt-fermented seahorse. In the enzyme-free group, it showed little change in the content of non-protein nitrogenous compounds, the content of amino acids and degree of hydrolysis. The Protamex enzyme treatment group was rapidly hydrolyzed in one week of ripening, resulting in increased non-protein nitrogenous compounds content, amino acid content and degree of hydrolysis, and minimal changes in the four weeks. The total amino acid contents ratio showed the highest content of glutamic acid in the enzyme additive group, glycine, alanine, which indicates sweet taste, and serine, the content of glycine, alanine, serine, and lysine, indicating sweet taste, has increased significantly over the enzyme-free group. Twenty species of free amino acid in the four-week of salt-fermented seahorse were detected. It detected 43.0% (6 species) in the enzyme-free group and 63.96% (7 species) in the enzyme additive group.

Comparison of taste ingredients between Long-term aged Korean Ganjang and manufactured brewed Ganjang

  • Kyung Tae JANG;In Sook LEE
    • The Korean Journal of Food & Health Convergence
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    • v.9 no.2
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    • pp.19-24
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    • 2023
  • In this study, we tried to compare and analyze the taste components of Korean Ganjang made by a conventional method on a small scale and aged for 3 years and brewed Ganjang produced by a large company. As a result of the study, a total of 22 types of free amino acids in liver were detected. The main amino acids of Korean Ganjang were glutamic acid, lysine, serine, citrulline, alanine, and leucine, then, the main amino acids of brewed Ganjang were leucine, glutamic acid, aspartic acid, valine, serine, alanine. The content of glutamic acid in the composition of free amino acid was 12133.69 mg/mL, more than twice as much as that of Korean Ganjang. The content of leucine (bitter taste) was 5933.37 mg/mL of brewed Ganjang, which was 2.9 to 7 times higher than that of Korean Ganjang. Overall, the content of glass amino acids with savory, sweet, and bitter flavors was found to be very high in brewed Ganjang (BGS) than in Korean Ganjang (KAS, KBS, and KCS). On the other hand, GABA was 456.43 mg/mL for Korean Ganjang KBS, 3.3 times higher than brewed Ganjang. Brewed Ganjang had higher glucose content, inorganic content, iron (Fe), organic acid lactic acid and acetic acid content than Korean Ganjang. Korean Ganjang was found to contain high saturated fatty acids, calcium (Ca), and sodium (Na).

Investigation of the effect of SRSF9 overexpression on HIV-1 production

  • Ga-Na, Kim;Kyung-Lee, Yu;Hae-In, Kim;Ji Chang, You
    • BMB Reports
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    • v.55 no.12
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    • pp.639-644
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    • 2022
  • Serine-arginine-rich splicing factors (SRSFs) are members of RNA processing proteins in the serine-arginine-rich (SR) family that could regulate the alternative splicing of the human immunodeficiency virus-1 (HIV-1). Whether SRSF9 has any effect on HIV-1 regulation requires elucidation. Here, we report for the first time the effects and mechanisms of SRSF9 on HIV-1 regulation. The overexpression of SRSF9 inhibits viral production and infectivity in both HEK293T and MT-4 cells. Deletion analysis of SRSF9 determined that the RNA regulation motif domain of SRSF9 is important for anti-HIV-1 effects. Furthermore, overexpression of SRSF9 increases multiple spliced forms of viral mRNA, such as Vpr mRNA. These data suggest that SRSF9 overexpression inhibits HIV-1 production by inducing the imbalanced HIV-1 mRNA splicing that could be exploited further for a novel HIV-1 therapeutic molecule.