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Enzymatic Preparation and Antioxidant Activities of Protein Hydrolysates from Protaetia brevitarsis Larvae (흰점박이꽃무지 유충 단백가수분해물의 제조 및 항산화 활성)

  • Lee, Hyo-Seon;Ryu, Hee-Jeong;Song, Hyeon-Ji;Lee, Syng-Ook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.10
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    • pp.1164-1170
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    • 2017
  • Protaetia brevitarsis larvae (PBL) has recently been registered as a temporary food in Korea, and this study evaluated the application potential of PBL proteins as health functional food materials. Protein hydrolysates were prepared from PBL powder by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain), and based on the results from the peptide content and SDS-PAGE analyses, PBL treated with alcalase or flavourzyme showed a high degree of hydrolysis (HD) value, whereas the HD value of those treated with neutrase, bromelain, or papain was minimal. The protein hydrolysates showing a high HD value were separated further into the fractions of >3 kDa and <3 kDa by a centrifugal filter system and then lyophilized, and according to the $RC_{50}$ values of the protein hydrolysates (<3 kDa) obtained from three different antioxidant analyses; the alcalase hydrolysates showed the highest antioxidant activity. Therefore, the alcalase hydrolysates were tested further for their inhibitory effects on the peroxidation of linoleic acid by measuring the thiobarbituric acid values. The results showed that the peroxidation of untreated linoleic acid increased dramatically during 6 days of incubation, but a pretreatment with the hydrolysates ($100{\sim}800{\mu}g/mL$) significantly inhibited the linoleic acid peroxidation in a dose-dependent manner for 6 days. Our current studies are focused on the identification of active peptide sequences from alcalase hydrolysates.

Enhancement of Skin Permeation of Wrinkle Improvement Peptides GHKs Using Liposomes Containing Skin Penetrating Peptides (피부 투과 펩티드가 함유된 리포좀을 이용한 주름 개선 펩티드 GHKs의 피부 흡수 증진)

  • Park, Su In;An, Gyu Min;Kim, Min Gi;Heo, Soo Hyeon;Shin, Moon Sam
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.3
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    • pp.853-865
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    • 2019
  • In this study, the skin permeability was measured by adding skin penetrating peptides, arginine oligomers R4(tetra-D-arginine), R6(hexa-D-arginine) to little skin-permeable wrinkle improvement peptides GHK, GHK-Cu, and Pal-GHK liposomes, and the results were analyzed by the following six cases. (1) In cases where only wrinkle improvement peptides GHK, GHK-Cu, and Pal-GHK were contained liposomes; the final cumulative permeations in 24 hours were 6.05%, 7.4%, and 8.83% respectively. (2) In cases where arginine oligomers R4, R6 were added to GHK liposomes; the final cumulative permeations in 24 hours were 13.63% and 7.68%. (3) In cases where R4, R6 were added to GHK-Cu liposomes; the final cumulative permeations in 24 hours were 15.46% and 8.64%. (4) In cases where R4, R6 were added to Pal-GHK liposomes; the final cumulative permeations in 24 hours were 16.9% and 10.67%. (5) In cases where R4 were added to GHK, GHK-Cu, and Pal-GHK liposomes; the final cumulative permeations in 24 hours were 13.63%, 15.46%, and 16.9% respectively. (6) In cases where R6 were added to GHK, GHK-Cu, and Pal-GHK liposomes; the final cumulative permeations in 24 hours were 7.68%, 8.64%, and 10.67% respectively. This experiment showed that skin absorption of GHK was increased by copper ion (Cu2+) and palmitic acid and skin absorption of wrinkle improvement peptides was enhanced by cell penetrating peptides, and R4 showed higher effect than R6 in GHK, GHK-Cu and Pal-GHK. Through this process, we propose broad use and application in wrinkle improvement functional cosmetics by presenting the optimal conditions for increasing skin absorption of GHK, GHK-Cu, thus maximizing its efficacy.

Inhibitory Effect of Protaetiamycine 6 on Neuroinflammation in LPS-stimulated BV-2 Microglia (LPS에 의해 활성화된 미세아교세포에서 흰점박이꽃무지 유래 항균 펩타이드 Protaetiamycine 6의 신경염증 억제 효과)

  • Lee, Hwa Jeong;Seo, Minchul;Baek, Minhee;Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.30 no.12
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    • pp.1078-1084
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    • 2020
  • Protaetia brevitarsis seulensis is an insect belonging to the order Coleoptera. This insect is reported to contain large amounts of physiologically active substances useful for liver protective effect and improvements in blood circulation as well as a broad source of edible protein. Antimicrobial peptides (AMPs) are found in a variety of species, from microorganisms to mammals, and play an important role in the innate immune systems of living things. Microglia are the main source of proinflammatory cytokines and nitric oxide (NO) in the central nervous system. Activated microglia secrete large amounts of neuroinflammatory mediators (e.g., TNF-α, NO, and ROS), which are the main cause of neuronal cell death. In the present study, we investigated the inhibitory effect of Protaetiamycine 6 (PKARKLQKLSAYKTTLRN-NH2), an AMP derived from Protaetia brevitarsis seulensis, on LPS-induced neuroinflammation in BV-2 microglia. Protaetiamycine 6 significantly inhibited NO production without cytotoxicity and decreased the expression levels of inducible NO synthase and cyclooxygenase-2. In addition, Protaetiamycine 6 also reduced the production of neuroinflammatory cytokines on activated BV-2 microglia. These results suggest that Protaetiamycine 6 could be a good source of functional substance to prevent neuroinflammation and neurodegenerative diseases.

Optimization of Peptides Production Derived from By-product Viscera of Yoensan Ogae Meat Process (연산 오계 부산물 내장육으로부터 펩타이드 생산 최적 공정 개발과 생산물의 특성 분석)

  • Ji, Joong Gu;Yoo, Sun Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.3
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    • pp.549-559
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    • 2016
  • Korean Black body fowl (Gallus gallus domesticus; Ogae) designated as a natural monument (registration number 265) has been known as a superb traditional Korean medicine. In this study, The production of peptide from the Viscera Waste of Yeonsan Ogae was optimized using commercial protease (bromelain) by response surface methodology under high pressure process. The range of processes was pressure (30 to 100 MPa), reaction time (1 to 5 h), and substrate concentration (10 to 30%, w/v). After reaction, the degree of hydrolysis, distribution of amino acids, and molecular weight of peptides were investigated. As a results, the optimization conditions were pressure 90 MPa, reaction time 3 to 4 h, and the amount of viscera meat 20% (w/v), respectively. The molecular weight of protein hydrolysates was distributed 400 to 1,000 Da. Accordingly we presumed that most products were peptides. Of those peptides, nonpolar or hydrophobic, polar but uncharged, positively charged, and negatively charged amino acids were 42.03, 26.0, 13.3, and 18.6%, respectively. Because higher amount of hydrophobic amino acids, we expected that those products would be able to utilize as the functional food ingredients.

Elucidation of the Biosynthetic Pathway of Vitamin B Groups and Potential Secondary Metabolite Gene Clusters Via Genome Analysis of a Marine Bacterium Pseudoruegeria sp. M32A2M

  • Cho, Sang-Hyeok;Lee, Eunju;Ko, So-Ra;Jin, Sangrak;Song, Yoseb;Ahn, Chi-Yong;Oh, Hee-Mock;Cho, Byung-Kwan;Cho, Suhyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.505-514
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    • 2020
  • The symbiotic nature of the relationship between algae and marine bacteria is well-studied among the complex microbial interactions. The mutual profit between algae and bacteria occurs via nutrient and vitamin exchange. It is necessary to analyze the genome sequence of a bacterium to predict its symbiotic relationships. In this study, the genome of a marine bacterium, Pseudoruegeria sp. M32A2M, isolated from the south-eastern isles (GeoJe-Do) of South Korea, was sequenced and analyzed. A draft genome (91 scaffolds) of 5.5 Mb with a DNA G+C content of 62.4% was obtained. In total, 5,101 features were identified from gene annotation, and 4,927 genes were assigned to functional proteins. We also identified transcription core proteins, RNA polymerase subunits, and sigma factors. In addition, full flagella-related gene clusters involving the flagellar body, motor, regulator, and other accessory compartments were detected even though the genus Pseudoruegeria is known to comprise non-motile bacteria. Examination of annotated KEGG pathways revealed that Pseudoruegeria sp. M32A2M has the metabolic pathways for all seven vitamin Bs, including thiamin (vitamin B1), biotin (vitamin B7), and cobalamin (vitamin B12), which are necessary for symbiosis with vitamin B auxotroph algae. We also identified gene clusters for seven secondary metabolites including ectoine, homoserine lactone, beta-lactone, terpene, lasso peptide, bacteriocin, and non-ribosomal proteins.

Bovine Growth Hormone and Milk Fat Synthesis: from the Body to the Molecule - Review -

  • Kim, W.Y.;Ha, J.K.;Han, In K.;Baldwin, R.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.4
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    • pp.335-356
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    • 1997
  • Injection of bovine growth hormone (bGH) to lactating dairy cows increases milk yield and yields of milk components including fat. It is generally believed that most of the anabolic effects derived from bGH in animal tissues are primarily mediated by IGF-1. IGF-1 is a strong anabolic peptide in the plasma of animals and exerts mitogenic and metabolic effects on target cells. Contrary to most protein hormones, the majority of IGF-1 in circulation is bound to the binding proteins (IGFBPs) which are known to be responsible for modifying the biological actions of IGF-1, thus making determinations of IGF-1 actions more difficult. On the other hand, fat is a major milk component and the greatest energy source in milk. Currently, the fat content of milk is one of the major criteria used in determining milk prices. It has been known that flavor and texture of dairy products are mainly affected by milk fat and its composition. Acetyl-CoA carboxylase (ACC) is the rate limiting enzyme which catalyzes the conversion of acetyl-CoA to malonyl-CoA for fatty acid synthesis in 1ipogenic tissues of animals including bovine lactating mammary glands. In addition to the short-tenn hormonal regulation of ACC by changes in the catalytic efficiency per enzyme molecule brought about by phosphorylation and dephosphorylation of the enzyme, the long-term hormonal regulation of ACC by changes in the number of enzyme molecules plays an essential role in control of ACC and lipogenesis. Insulin, at supraphysiological concentrations, binds to IGF-1 receptors, thereby mimicking the biological effects of IGF-1. The receptors for insulin and IGF-1 share structural and functional homology. Furthermore, epidermal growth factor increased ACC activity in rat hepatocytes and adipocytes. Therefore, it can be assumed that IGF-1 mediating bGH action may increase milk fat production by stimulation ACC with phosphorylation (short term) and/or increasing amounts of the enzyme proteins (long term). Consequently, the main purpose of this paper is to give the readers not only the galactopoietic effects of bGH, but also the insight of bGH action with regard to stimulating milk fat synthesis from the whole body to the molecular levels.

Effect of Dietary Soybean Hydrolysate on Plasma Lipid Profiles, Select Biochemical Indexes, and Histopathological Changes in Spontaneously Hypertensive Rats (대두가수분해물 유래 펩타이드 투여가 자발성 고혈압 흰쥐의 혈중 지질수준, 생화학적 지표 및 조직병리학적 변화에 미치는 영향)

  • Chung, Sun-Wha;Choi, Min-Ah;Park, Jeong-Soon;Kim, Kil-Soo;Chung, Dae-Kyun;Nam, Hee-Sop;Shin, Zae-Ik;Yu, Ri-Na
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.1101-1108
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    • 1999
  • Soybean (SoyPep) was prepared and evaluated their action on blood lipid profiles, select biochemical indexes and histopathological changes in spontaneously hypertensive rat (SHR). Male SHRs were divided into three groups and fed the experimental diet containing 0% (control), 3%, 10% SoyPep for 8 weeks. Lipids, antioxidative vitamin levels, glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) activities in plasma were measured, and angiotensin I converting enzyme (ACE) activities in kidney, vascular tissue and plasma were also measured. In addition, histopathological changes in various organs were investigated. Dietary SoyPep was effective for lowering plasma levels of triglyceride and LDL-cholesterol. ACE activities in aorta and kidney were decreased in the group fed SoyPep compared to control. There was no difference of plasma GOT and GPT activities, and no histopathological difference in various organs among the experimental groups. Our results suggest that SoyPep has beneficial effects on diseases such as hypertension and atherosclerosis. SoyPep could be useful to develop a novel functional safe food additive for preventing the chronic diseases.

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EST Analysis system for panning gene

  • Hur, Cheol-Goo;Lim, So-Hyung;Goh, Sung-Ho;Shin, Min-Su;Cho, Hwan-Gue
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2000.11a
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    • pp.21-22
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    • 2000
  • Expressed sequence tags (EFTs) are the partial segments of cDNA produced from 5 or 3 single-pass sequencing of cDNA clones, error-prone and generated in highly redundant sets. Advancement and expansion of Genomics made biologists to generate huge amount of ESTs from variety of organisms-human, microorganisms as well as plants, and the cumulated number of ESTs is over 5.3 million, As the EST data being accumulate more rapidly, it becomes bigger that the needs of the EST analysis tools for extraction of biological meaning from EST data. Among the several needs of EST analyses, the extraction of protein sequence or functional motifs from ESTs are important for the identification of their function in vivo. To accomplish that purpose the precise and accurate identification of the region where the coding sequences (CDSs) is a crucial problem to solve primarily, and it will be helpful to extract and detect of genuine CD5s and protein motifs from EST collections. Although several public tools are available for EST analysis, there is not any one to accomplish the object. Furthermore, they are not targeted to the plant ESTs but human or microorganism. Thus, to correspond the urgent needs of collaborators deals with plant ESTs and to establish the analysis system to be used as general-purpose public software we constructed the pipelined-EST analysis system by integration of public software components. The software we used are as follows - Phred/Cross-match for the quality control and vector screening, NCBI Blast for the similarity searching, ICATools for the EST clustering, Phrap for EST contig assembly, and BLOCKS/Prosite for protein motif searching. The sample data set used for the construction and verification of this system was 1,386 ESTs from human intrathymic T-cells that verified using UniGene and Nr database of NCBI. The approach for the extraction of CDSs from sample data set was carried out by comparison between sample data and protein sequences/motif database, determining matched protein sequences/motifs that agree with our defined parameters, and extracting the regions that shows similarities. In recent future, in addition to these components, it is supposed to be also integrated into our system and served that the software for the peptide mass spectrometry fingerprint analysis, one of the proteomics fields. This pipelined-EST analysis system will extend our knowledge on the plant ESTs and proteins by identification of unknown-genes.

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[ $P2X_2$ ] Receptor Activation Potentiates PC12 Cell Differentiation Induced by ACAP in Acidic Environments

  • Lee, Myung-Hoon;Nam, Jin-Sik;Ryu, Hye-Myung;Yoo, Min;Lee, Moon-Hee
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.197-206
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    • 2007
  • P2X receptors are membrane-bound ion channels that conduct $Na^+,\;K^+$, and $Ca^{2+}$ in response to ATP and its analogs. There are seven subunits identified so far ($P2X_1-P2X_7$). $P2X_2$ receptors are known to be expressed in a wide range of organs including brains and adrenal grands. PC12 cells are originated from adrenal grand and differentiated by nerve growth factor or pituitary adenylate cyclase activating poly peptide (PACAP). Previous studies indicate that $P2X_2$ receptor activation in PC12 cells couples to $Ca^{2+}-dependent$ release of catecholamine and ATP. It is known that acidic pH potentiates ATP currents at $P2X_2$ receptors. This leads to a hypothesis that $P2X_2$ receptors may play an important role in PC12 cell differentiation, one of the characteristics of which is neurite outgrowth, induced by the hormones under lower pH. In the present study, we isolated several clones which potentiate neurite outgrowth by PACAP in acidic pH (6.8), but not in alkaline pH (7.6). RT-PCR and electrophysiology data indicate that these clones express only functional $P2X_2$ receptors in the absence or presence of PACAP for 3 days. Potentiation of neurite outgrowth resulted from PACAP (100 nM) in acidic pH is inhibited by the two P2X receptor antagonists, suramin and PPADS ($100\;{\mu}M)$ each), and exogenous exprerssion of ATP-binding mutant $P2X_2$ receptor subunit ($P2X_2[K69A]$). However, acid sensing ion channels (ASICs) are not involved in PACAP-induced neurite outgrowth potentiation in lower pH since treatments of an inhibitor of ASICs, amyloride ($10\;{\mu}M$), did not give any effects to neurite extension. The vesicular proton pump ($H^+-ATPase$) inhibitor, bafilomycin (100 nM), reduced neurite extension indicating that ATP release resulted from $P2X_2$ receptor activation in PC12 cells is needed for neurite outgrowth. These were confirmed by activation of mitogen activated protein kinases, such as ERKs and p38. These results suggest roles of ATP and $P2X_2$ receptors in hormone-induced cell differentiation or neuronal synaptogenesis in local acidic environments.

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Are Biomarkers Predictive of Anthracycline-Induced Cardiac Dysfunction?

  • Malik, Abhidha;Jeyaraj, Pamela Alice;Calton, Rajneesh;Uppal, Bharti;Negi, Preety;Shankar, Abhishek;Patil, Jaineet;Mahajan, Manmohan Kishan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2301-2305
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    • 2016
  • Background: The early detection of anthracycline- induced cardiotoxicity is very important since it might be useful in prevention of cardiac decompensation. This study was designed with the intent of assessing the usefulness of cardiac troponin T (cTnT) and NT- Pro BNP estimation in early prediction of anthracycline induced cardiotoxicity. Materials and Methods: In this prospective study histologically proven breast cancer patients who were scheduled to receive anthracycline containing combination chemotherapy as a part of multimodality treatment were enrolled. Baseline cardiac evaluation was performed by echocardiography (ECHO) and biomarkers like cardiac troponin T (cTnT) and N terminal- pro brain natriuretic peptide (NT- Pro BNP). All patients underwent cTnT and NT- Pro BNP estimation within 24 hours of each cycle of chemotherapy and were followed up after 6 months of initiation of chemotherapy. Any changes in follow up ECHO were compared to ECHO at baseline and cTnT and NT- Pro BNP levels after each cycle of anthracycline-based chemotherapy. Results: Initial data were obtained for 33 patients. Mean change in left ventricular diastolic diameter (LVDD) within 6 months was $0.154{\pm}0.433cms$ (p value=0.049). Seven out of 33 patients had an increase in biomarker cTnT levels (p value=0.5). A significant change in baseline and follow up LVDD was observed in patients with raised cTnT levels (p value=0.026) whereas no change was seen in ejection fraction (EF) and left atrial diameters (LAD) within 6 months of chemotherapy. NT- Pro BNP levels increased in significant number of patients (p value ${\leq}0.0001$) but no statistically significant change was observed in the ECHO parameters within 6 months. Conclusions: Functional monitoring is a poorly effective method in early estimation of anthracycline induced cardiac dysfunction. Estimation of biomarkers after chemotherapy may allow stratification of patients in various risk groups, thereby opening window for interventional strategies in order to prevent permanent damage to the myocardium.