• Title/Summary/Keyword: protease inhibition

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Effect of Solid-State Fermented Brown Rice Extracts on 3T3-L1 Adipocyte Differentiation

  • Su Bin Ji;Chae Hun Ra
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.926-933
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    • 2023
  • Aspergillus oryzae KCCM 11372 was used to enhance the production of β-glucan using humidity control strategies. Under conditions of 60% humidity, solid-state fermentation (SSF) increased the yields of enzymes (amylase and protease), fungal biomass (ergosterol), and β-glucan. The maximum concentrations obtained were 14800.58 U/g at 72 h, 1068.14 U/g at 120 h, 1.42 mg/g at 72 h, and 12.0% (w/w) at 72 h, respectively. Moreover, the β-glucan containing fermented brown rice (β-glucan-FBR) extracts at concentrations of 25-300 ㎍/ml was considered noncytotoxic to 3T3-L1 preadipocytes. We then studied the inhibitory effects of the extracts on fat droplet formation in 3T3-L1 cells. As a result, 300 ㎍/ml of β-glucan-FBR extracts showed a high inhibition of 38.88% in lipid accumulation. Further, these extracts inhibited adipogenesis in the 3T3-L1 adipocytes by decreasing the expression of C/EBPα, PPARγ, aP2, and GLUT4 genes.

Characteristic of Glasswort (Salicornia Herbacea L.) Mixture Fermentation Utilizing Aspergillus oryzae (황국균을 이용한 퉁퉁마디(Salicornia herbacea L.) 혼합물의 발효 특성)

  • Kim, Hae-Seop;Park, In-Bae;Lee, Young-Jae;Shin, Gung-Won;Lim, Joo-Young;Park, Jeong-Wook;Jo, Young-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.9
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    • pp.1384-1390
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    • 2010
  • In this study, we investigated the quality of glasswort (Salicornia herbacea L.) mixture fermented by Aspergillus oryzae at $30^{\circ}C$ for 7 days. Changes of pH, titratable acidity, amino-nitrogen content, reducing sugar content, activities of $\alpha$-amylase and protease and number of mold were determined in the course of the fermentation. Angiotensin converting enzyme (ACE) inhibition activities and electron donating ability (EDA) were measured after 7 days fermentation. The pH lowered from 6.19~6.22 into the level of 5.41~6.08 after fermentation for 7 days. Titratable acidity increased from 0.59~0.68 into the highest level of 0.95~1.13% after 6 days fermentation. Furthermore, the amino-nitrogen content increased from 128.0~167.2 mg% to 159.4~209.0 mg% after fermentation for 7 days. For the reducing sugar content, it decreased from 2.0~5.9% into the level of 0.4~1.1% during 7 days fermentation. The number of molds decreased from $10^7\;CFU/g$ to approximately $10^6\;CFU/g$ after 5 days fermentation. $\alpha$-Amylase activity showed from the beginning of the fermentation in all samplings, but protease activity reached the value of food code standards after 5 days of fermentation. ACE inhibition activities were slightly increased from 9.5~16.6% to 19.3~22.7% and electron donating ability were slightly increased from 55.6~57.8% to 60.9~62.7% after 7 days of fermentation.

Physiological Function and Enzyme Activity of Koji Cultured by Aspergillus oryzae CNU O4-5 in the Various Grain Materials (원료(原料)의 종류(種類)와 형태(形態)에 따른 Aspergillus oryzae 국(麴)의 생리활성(生理活性) 및 효소력(酵素力))

  • Choi, Seong Hyun;Jeong, Jae-Hong;Lee, Sang Duck;Son, Hwa Young;Sung, Chang;Oh, Man Jin;Kim, Seung Yeol
    • Korean Journal of Agricultural Science
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    • v.24 no.2
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    • pp.283-289
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    • 1997
  • CNU O4-5 isolated from conventional Meju, which is used as raw material for making a soybean fermentation food, identified as an Aspergillus oryzae. To make koji, Aspergillus oryzae CNU O4-5 was cultured for 3-4 days at $30^{\circ}C$ with various grain materials such as flour, soybean powder, flour+soybean powder(1:1), soaked soybean and rice. The koji was evaluated for analyze the angiotesin converting enzyne(ACE) inhibition, antioxidative activity, superoxide dismutase(SOD) activity, amylase and protease activity. $\alpha$-amylase and glucoamylase activities of flour koji were higher than those of the koji soybean powder. However neutral and alkaline protease activities of flour koji were lower than those of flour+soybean powder and soybean koji. Amylase and protease activities of kojies of soaked soybean and rice showed very low level. The range of the ACE inhibition rate by hot water fraction of the kojies, which are cultured with various gain materials, were from 45% to 54%. The anti oxidative activity of ethanol-fraction of koji, which is made by using the soybean powder or soaked soybean, prolonged for 6 days in lard at $60^{\circ}C$. The SOD activity of grinded fraction of koji, which is made by using the flour or soybean powder, was same as 2,000 units per g of each koji.

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Some Physiological Activity of Phenolic Substances in Plant Foods (식물성 식품중 페놀성 물질의 몇가지 생리활성)

  • Lee, Jung-Hi;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.26 no.3
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    • pp.317-323
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    • 1994
  • Nine plant foods (persimmon leaf, perilla seed, Chinese quince, ginger root, walnut, mugwort leaf, arrowroot, buckwheat and sorghum) rich in phenolic substances were examined for their effects on the digestive enzymes, food-poisoning bacteria and mutagenicity/antimutagenicity by Ames test. Among tested samples, Chinese quince significantly inhibited the $\alpha-amylase$ activity (97%), exhibiting an uncompetitive inhibition type. Protease activity was inhibited by Chinese quince (86%), persimmon leaf (51%) and mugwort leaf (20%), in which mugwort extract exhibited a noncompetitive type. Lipase was activated >50% by all samples. The inhibition of $\alpha-amylase$ was highly correlated with the content of condensed tannin (r=0.89) and the inhibition of protease, with total phenolic content (r=0.84). Total phenolies fraction of tested samples showed the growth inhibition toward E. coli. Streptococcus faecalis and Salmonella enteritidis, in which the effect of perilla, sorghum and arrowroot was the highest for E. coli. Standard phenolics and food samples did not show any mutagenicity toward Salmonella typhimurium TA98 and TA100. Tannic acid inhibited the mutation of the two strains by benzo[a]pyrene whereas total phenolics fractions of Chinese quince and walnut exhibited antimutagenicity to a lesser extent.

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Antioxidant Activity of Ogae Egg White Protein Hydrolysates using commercial Protease (단백질 분해효소를 이용한 오계란 가수분해물의 항산화 활성)

  • Ha, Yoo Jin;Ji, Joong Gu;Yoo, Sun Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.631-642
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    • 2017
  • Protein hydrolysates derived from plants and animals having antioxidant, suppression of hypertension, immunodulatory, alleviation of pain, and antimicrobial activity has been known as playing important role like hormone. This study was performed to hydrolysis of Ogae egg white protein using the six proteases. The antioxidant activity of the produced peptides was analyzed. As a result, the maximum value of hydrolysis was protamex(46.3%), DPPH radical scavenging was bromelain(57.23%), hydroxy radical scavenging was alcalase(30.21%), superoxide radical scavenging was alcalase(58.07%), and $Fe^{2+}$ chelation ability was alcalase(72.06%). Furthermore, the antioxidant Inhibition concentration ($IC_{50}$) of peptides was evaluated for each enzyme. As a result, the maximum value of alcalase was $Fe^{2+}$ cheating ability($IC_{50}$, 1.24 mg/mL), bromelain was DPPH radical scavenging($IC_{50}$, 2.46 mg/mL), flavourzyme was $Fe^{2+}$ cheating ability($IC_{50}$, 1.25 mg/mL), neutrase was DPPH radical scavenging($IC_{50}$, 3.64 mg/mL), papain was DPPH radical scavenging ($IC_{50}$, 3.82 mg/mL) and protamex was DPPH radical scavenging($IC_{50}$, 1.93 mg/mL). Therefore, we expect that peptides produced from Ogae egg white protein using protease enzyme are useful as an antioxidant functional food ingredients.

Production and Separation of Anti-hypertensive Peptide during Chunggugjang Fermentation with Bacillus subtilis CH-1023 (청국장 발효과정 중 항고혈압성 peptide의 생산 및 분리)

  • Cha, Woen-Suep;Bok, Su-Kyung;Kim, Myoung-Uk;Chun, Sung-Sook;Choi, Ung-Kyu;Cho, Young-Je
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.247-252
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    • 2000
  • As functionality investigation of Korean traditional soybean fermentation foods, an antihypertensive peptide was separated during Chunggugjang fermentation by Bacillus subtilis CH-1023 and investigated inhibitory effect against angiotensin converting enzyme. After incubation at $20^{\circ}C,\;30^{\circ}C,\;40^{\circ}C,\;50^{\circ}C,\;60^{\circ}C$ for the $0{\sim}72$ hrs, protein content, protease activity and angiotensin converting enzyme inhibitory rate were determined. The protein content and protease activity were increased and reached maximum at 60 hrs fermentation with $40^{\circ}C$ and decreased after the 60 hrs fermentation. The optimum condition for antihypertensive peptide from Chunggugjang was appeared for 60 hrs at $40^{\circ}C$. Crude extract of Chunggugjang was partially purified by Amicon YM-3 membrane filtration and Sephadex G-10, G-25 gel filtration. The purified peptide showed inhibitory rate of 94.3% with 0.5 mg peptide content. The most prominent amino acid composition of the peptide from Chunggugjang was alanine, followed by phenylalanine, histidine.

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Production and Separation of Angiotension Converting Enzyme Inhibitor during Natto Fermentation (납두 발효과정 중 Angiotensin Converting Enzyme 저해물질의 생성 및 분리)

  • Cho, Young-Je;Cha, Woen-Suep;Bok, Su-Kyung;Kim, Myung-Uk;Chun, Sung-Sook;Choi, Ung-Kyu;Kim, Soon-Hee;Park, Kyung-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.737-742
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    • 2000
  • As functionality investigation of a soybean fermentation food, a angiotensin converting enzyme inhibitory peptide was separated during natto fermentation by Bacillus natto and inhibitory effect was investigated. After incubation at each 2$0^{\circ}C$, 3$0^{\circ}C$, 4$0^{\circ}C$, 5$0^{\circ}C$, 6$0^{\circ}C$ for the 0~72 hr, protein content, protease activity and angiotensin converting enzyme inhibition were determined. The protein content and protease activity were increased and reached maximum at 60 hr fermentation with 4$0^{\circ}C$ and decreased after the 60 hr fermentation during natto fermentation. The optimum condition for angiotensin converting enzyme inhibitors was appeared at fermentation for 60 hr at 4$0^{\circ}C$. Crude extract of natto was partially purified by Amicon membrane YM-3 and Sephadex G-10, G-25 gel filtration, stepwise. The inhibitory rate was increased in a concentration dependent manner, espcially the most potent activity about 74.74% at 1.0 mg peptide content. The most prominent amino acid of the peptide from natto was alanine, followed by phenylalnine, histidine.

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Curcumin-Induced Apoptosis of A-431 Cells Involves Caspase-3 Activation

  • Shim, Joong-Sup;Lee, Hyung-Joo;Park, Sang-shin;Cha, Bong-Gee;Chang, Hae-Ryong
    • BMB Reports
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    • v.34 no.3
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    • pp.189-193
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    • 2001
  • Curcumin a yellow pigment from Curcuma Tonga, has been known to possess antioxidative and anticarcinogenic properties, as well as to induce apoptosis in some cancer cells. There have been, however, several contradictory reports that hypothesized curcumin (a hydrophobic molecule) can bind a membrane Gpid bilayer and induce nonspecific cytotoxicity in some cell lines. Why curcumin shows these contradictory effects is unknown. In A-431 cells, growth inhibition by curcumin is due mostly to the specific inhibition of the intrinsic tyrosine kinase activity of the epidermal growth factor receptor, as reported earlier by Korutla et al. Thus, we assumed that the cell death of A-431 by curcumin might be due to the specific induction of apoptosis. In this paper we clearly show that curcumin induces apoptosis in A-431 cells. The cureumin-induced cell death of A-431 exhibited various apoptotic features, including DNA fragmentation and nuclear condensation. Furthermore, the curcumin-induced apoptosis of A-431 cells involved activation of caspase-3-like cysteine protease. Involvement of caspase-3 was further confirmed by using a caspase-3 specific inhibitor, DEVD-CHO. In another study, decreased nitric oxide (NO) production was also shown in A-431 cells treated with curcumin, which seems to be the result of the inhibition of the iNOS expression by curcumin, as in other cell lines. However, 24 h after treatment of curcumin there was increased NO production in A-431 cells. This observation has not yet been clearly explained. We assumed that the increased NO production may be related to denitrosylation of the enzyme catalytic site in caspase-3 when activated. Taken together, this study shows that the cell death of A-431 by curcumin is due to the induction of apoptosis, which involves caspase-3 activation.

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Overexpression of three related root-cap outermost-cell-specific C2H2-type zinc-finger protein genes suppresses the growth of Arabidopsis in an EAR-motif-dependent manner

  • Song, Sang-Kee;Jang, Hyeon-Ung;Kim, Yo Han;Lee, Bang Heon;Lee, Myeong Min
    • BMB Reports
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    • v.53 no.3
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    • pp.160-165
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    • 2020
  • The root meristem of Arabidopsis thaliana is protected by the root cap, the size of which is tightly regulated by the balance between the formative cell divisions and the dispersal of the outermost cells. We isolated an enhancer-tagged dominant mutant displaying the short and twisted root by the overexpression of ZINC-FINGER OF ARABIDOPSIS THALIANA1 (ZAT1) encoding an EAR motif-containing zinc-finger protein. The growth inhibition by ZAT1 was shared by ZAT4 and ZAT9, the ZAT1 homologues. The ZAT1 promoter was specifically active in the outermost cells of the root cap, in which ZAT1-GFP was localized when expressed by the ZAT1 promoter. The outermost cell-specific expression pattern of ZAT1 was not altered in the sombrero (smb) or smb bearskin1 (brn1) brn2 accumulating additional root-cap layers. In contrast, ZAT4-GFP and ZAT9-GFP fusion proteins were distributed to the inner root-cap cells in addition to the outermost cells where ZAT4 and ZAT9 promoters were active. Overexpression of ZAT1 induced the ectopic expression of PUTATIVE ASPARTIC PROTEASE3 involved in the programmed cell death. The EAR motif was essential for the growth inhibition by ZAT1. These results suggest that the three related ZATs might regulate the maturation of the outermost cells of the root cap.

Extract of Linum usitatissimum L. inhibits Coxsackievirus B3 Replication through AKT Signal Modulation (아마인 추출물의 AKT 신호 조절을 통한 콕사키바이러스 증식억제)

  • Shin, Ha-Hyeon;Moon, Sung-Jin;Lim, Byung-Kwan;Kim, Jin Hee
    • Korean Journal of Pharmacognosy
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    • v.49 no.4
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    • pp.291-297
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    • 2018
  • Coxsackievirus B3 (CVB3) is a very well-known causative agent for viral myocarditis and meningitis in human. However, the effective vaccine and therapeutic drug are not developed yet. CVB3 infection activates host cell AKT signaling. Inhibition of AKT signaling pathway may attenuate CVB3 replication and prevent CVB3-mediate viral myocarditis. In this study, we determined antiviral effect of the selected natural plant extract to develop a therapeutic drug for CVB3 treatment. We screened several chemically extracted natural compounds by using HeLa cell-based cell survival assay. Among them, Linum usitatissimum L. extract was selected for antiviral drug candidate. L. usitatissimum extract significantly decreased CVB3 replication and cell death in CVB3 infected HeLa cells with no cytotoxicity. CVB3 protease 2A induced eIF4G1 cleavage and viral capsid protein VP1 production were dramatically decreased by L. usitatissimum extract treatment. In addition, virus positive and negative strand genome amplification were significantly decreased by 1 mg/ml L. usitatissimum extract treatment. Especially, L. usitatissimum extract was associated with inhibition of AKT signal and maintain mTOR activity. In contrast, Atg12 and LC3 expression were not changed by L. usitatissimum extract treatment. In this study, the potential AKT signal inhibitor, L. usitatissimum extract, was significantly inhibited viral genome replication and protein production by inhibition of AKT signal. These results suggested that L. usitatissimum extract is a novel therapeutic agent for treatment of CVB3-mediated diseases.