• Title/Summary/Keyword: protease production

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Production of Selenium Peptide by Autolysis of Saccharomyces cerevisiae

  • Lee Jung-Ok;Kim Young-Ok;Shin Dong-Hoon;Shin Jeong-Hyun;Kim Eun-Ki
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1041-1046
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    • 2006
  • Selenium-containing peptide (selenium peptide) was produced by autolysis of total proteins of Saccharomyces cerevisiae grown with inorganic selenium. Selenium peptide exhibited antioxidant activity as a glutathione peroxidase (GPx) mimic, and its activity was dependent on the hydrolysis methods. The GPx-like activity of the hydrolyzed selenium peptide increased 2.7-folds when digested by protease, but decreased by acid hydrolysis. During the autolysis of the yeast cell, the GPx-like activity and selenium content increased 4.3- and 2.3-folds, respectively, whereas the average molecular weight (MW) of selenium peptide decreased 70%. The GPx-like activity was dependent on the MW of selenium peptide and was the highest (220 U/mg protein) at 9,500 dalton. The maximum GPx-like activity (28,600 U/g cell) was obtained by 48 h of autolysis of the cells, which were precultured with 20 ppm of selenate. Selenium peptide showed little toxicity, compared with highly toxic inorganic selenium. These results show the potential of selenium peptide as a nontoxic antioxidant that can be produced by simple autolysis of yeast cells.

The Effects of DL-methionine and DL-methionine Hydroxy Analogue on Growth Performance, Contents of Serum Amino Acids and Activities of Digestive Proteases in Broilers

  • Lu, J.J.;Huang, C.W.;Chou, R.G.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.714-718
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    • 2003
  • This study was conducted to determine the effect of DL-methionine (Met) and DL-methionine hydroxy analogue (MHA) supplementation on the growth performance, contents of serum amino acids and activities of digestive protease in broiler chicks from 0 to 3 weeks old . There were three treatments in this study: (1) control (basal diet), (2) 0.24% Met supplementation and (3) 0.368% MHA supplementation. The results showed that Met and MHA supplementation did not significantly (p>0.05) improve feed efficiency and body weight gain for broilers from 0-1, 0-2 and 0-3 weeks of age. The serum levels of homocystine, methionine and taurine were significantly (p<0.05) higher with supplementation of Met or MHA than with control. The pepsin activity of proventriculus was increased with (p<0.05) Met supplementation at 21 days of age and with MHA supplementation at 7 and 14 days of age. The trypsin activity was also increased (p<0.05) with MHA supplementation at 7 days of age. The chymotrypsin activity in pancreas and the dipeptidase activity in small intestinal mucosa and content were not affected (p>0.05) by Met or MHA supplementation.

Comparison of Digestive Function Among Rabbits, Guinea-Pigs, Rats and Hamsters. II. Digestive Enzymes and Hindgut Fermentation

  • Yu, Bi;Chiou, Peter Wen-Shyg;Kuo, Chung-Yi
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.11
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    • pp.1508-1513
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    • 2000
  • The aim of this trial was to study the response of laboratory animals including omnivores (rats) and herbivores (rabbits, guinea pigs and hamsters) to the same level of dietary fiber on their digestive enzymes and hindgut fermentation. Ten weanling animals of each of four species, rabbits, guinea-pigs, rats and Syrian hamster, were fed a basal diet of 18% crude protein and 10% crude fiber for six weeks. The digesta and tissue of each intestinal segment were collected to measure the activity of digestive enzymes. Rabbits contained the highest secreted pepsin activity in the stomach, whereas rats contained the highest protease and ${\alpha}-amylase$ activity in the small intestine, and lower fibrous hydrolases in the hindgut than rabbits, guinea pigs and hamsters. The total VFA productions in the caecum and colon were highest in rats, followed by hamsters and rabbits, while the guinea pigs contained the lowest VFA and a different pattern of VFA molar ratio from the other laboratory animals. The degree of hindgut fermentation in these laboratory animals was in reverse to the trend for their fiber digestion.

재래식 메주에서 분리한 효모들의 각종 효소활성과 가능성

  • Lee, Jong-Soo;Yi, Sung-Hun;Kwon, Su-Jin;Ahn, Cheol;Yoo, Jin-Young
    • Microbiology and Biotechnology Letters
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    • v.25 no.5
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    • pp.448-453
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    • 1997
  • Enzyme activities, production of killer toxin and some functionality of forty seven yeasts isolated from traditional Meju were investigated in culture broth and cell free extracts. Activities of $\alpha$-galactosidase, invertase and inulinase were detected in cell free extracts of 38 strains, 43 strains and 45 strains, respectively and acidic and neutral protease activities also were detected in culture broth of all the strains, $\beta$-Galactosidase activity was detected in cell free extracts of OE-20 and S-14 strains. Killer toxins were produced by OE-12, S-8 (Candida spp.), OE-19 (Zygosaccharomyces spp.) and S-3 (Saccharomyces spp.). Culture broth of OE-23 and S-9 showed 61.3% and 59.2% of antioxidant activity to $\alpha$, $\alpha$-diphenyl-$\beta$-picrylhydrazyl(DPPH), but nitrite-scavenging ability as well as inhibition of tyrosinase and polyphenol oxidase were not appeared in all the strains.

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Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage

  • Lee, Kang Pa;Kim, Jai-Eun;Park, Won-Hwan
    • Nutrition Research and Practice
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    • v.9 no.6
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    • pp.586-591
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    • 2015
  • BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount$^{TM}$ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2',7'-dichlorofluoroscein diacetate ($H_2DCF$-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and $10{\mu}M$) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole ($3{\mu}M$)-treated cells in a dose-dependent manner. Rhamnetin (1 and $3{\mu}M$) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.

Changes in Microflora and Enzyme Activities of Traditional Meju during Fermentation at Sunchang Area (순창지역 메주 발효 중 미생물과 효소역가의 변화)

  • 유진영;김현규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.3
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    • pp.448-454
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    • 1998
  • Meju for doenjang and kochujang was prepared as a model at Sunchang areaand monitored with major changes. Kochujang meju was prepared on September 12 and doenjang meju on November 12, 1995. Kochjang meju was found to be naturally fermented at 80~90% RH, 15~2$0^{\circ}C$ and doenjang meju was at 80~90% RH and 0~5$^{\circ}C$. The shapes of kochujang meju and doenjang meju were doughnut-type and rectangular, respectively. Weight losses during fermentation were 48% and 28%, respectively. The pH drop and acip production of kochujang meju were negligible. However, pH of doenjang meju decreased from 6.29 to 5.88 and acidity increased from 0.08 to 0.23% as lactic acid. Protein in meju was found to be rapidly solubilized during the early stage of fermentation. Soluble protein cotents of kochujang meju after 7 days and 60 days were 8.23%, respectively. The doenjang mejus were 2.15% after 20 days and 5.72% after 60 days. Soluble suger content increased with the fermentation time. The soluble sugar content was higher in kochjang meju. Acidic protease was highly produced during meju fermentation. $\alpha$-Amylase and $\beta$-amylase were detected in the kochujang meju, of which glutinous rice consisted, but negligible in doenjang meju. Lipase was detected in kochujang meju, but was, negligible in doenjang meju. Microbial population increased drastically after 7 days of fermentation in kochujang meju and 20 days of fermentation in doenjang meju.

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Biological Control of Apple Anthracnose by Paenibacillus polymyxa APEC128, an Antagonistic Rhizobacterium

  • Kim, Young Soo;Balaraju, Kotnala;Jeon, Yongho
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.251-259
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    • 2016
  • The present study investigated the suppression of the disease development of anthracnose caused by Colletotrichum gloeosporioides and C. acutatum in harvested apples using an antagonistic rhizobacterium Paenibacillus polymyxa APEC128 (APEC128). Out of 30 bacterial isolates from apple rhizosphere screened for antagonistic activity, the most effective strain was APEC128 as inferred from the size of the inhibition zone. This strain showed a greater growth in brain-heart infusion (BHI) broth compared to other growth media. There was a reduction in anthracnose symptoms caused by the two fungal pathogens in harvested apples after their treatment with APEC128 in comparison with non-treated control. This effect is explained by the increased production of protease and amylase by APEC128, which might have inhibited mycelial growth. In apples treated with different APEC128 suspensions, the disease caused by C. gloeosporioides and C. acutatum was greatly suppressed (by 83.6% and 79%, respectively) in treatments with the concentration of $1{\times}10^8$ colony forming units (cfu)/ml compared to other lower dosages, suggesting that the suppression of anthracnose development on harvested apples is dose-dependent. These results indicated that APEC128 is one of the promising agents in the biocontrol of apple anthracnose, which might help to increase the shelf-life of apple fruit during the post-harvest period.

The Effect of Poria cocos Extract to Inhibit Enterovirus Replication (적복령 추출물의 심근염 유발 엔테로바이러스 증식 억제 효과)

  • Han, Jae-Young;Kim, Jin Hee;Lim, Byung-Kwan
    • Korean Journal of Pharmacognosy
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    • v.47 no.2
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    • pp.137-142
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    • 2016
  • Enterovirus is a common cause of several severe diseases such as myocarditis, hand-foot-mouth disease, and meningitis in children and adult. There are many try to develop new antiviral drug for direct treatment in virus infection. However, synthetic chemical antiviral drug is not working. To overcome this limitation, we examined plant extracts. The antiviral effect of plant extracts was screened by HeLa cell survival assay in coxsackievirus B3 (CVB3) infection. We observed a strong antiviral effect of Poria cocos extract in a dose-dependent manner (1 mg/ml~0.01 mg/ml). P. cocos extract (1 mg/ml) treatment was dramatically decreased virus protease 2A induced eIF4G-I cleavage and virus capsid protein VP1 production. CVB3 positive and negative strand RNA amplification were significantly reduced in P. cocos extract treatment. P. cocos extract completely blocked early time activation of ERK and AKT activity in CVB3 infection. Taken together these data indicate that the treatment of P. cocos extract strongly inhibit CVB3 replication. Poria cocos extract may possible to developed as a therapeutic agent for enterovirus.

Laboratory Culture Media-Dependent Biocontrol Ability of Burkholderia gladioli strain B543

  • Bae, Yeoung-Seuk;Park, Kyung-Seok;Choi, Ok-Hee
    • The Plant Pathology Journal
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    • v.23 no.3
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    • pp.161-165
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    • 2007
  • Cultivation of a biocontrol agent on a certain medium often results in reduced biocontrol efficacy and alters physiological state. In our previous study, Burkholderia gladioli strain B543 with long-term subculture on tryptic soy agar resulted in significantly reduced biocontrol ability against cucumber damping-off caused by P. ultimum. Therefore, we investigated the influence of laboratory culturing media on biocontrol activity and physiological state of Burkholderia gladioli strain B543 by using long-term repeated culture on a certain medium. When isolate B543 were successionally cultured on King's B agar (KBA), tryptic soy agar, nutrient agar (NA), or soil extract agar more than 20 times, the isolate cultured on KBA or NA showed a significantly enhanced biocontrol efficacy and higher population density in the rhizosphere of cucumber compared to that of the others. However, the isolates cultured on KBA more than 20 times showed the lowest production of protease, siderophore, or antifungal substance(s), measured by skim milk agar, Chrome-Azurol-S agar, and potato dextrose agar amended with 10% of the culture filtrate, respectively. Our results suggest that adaptation to proper culturing medium can alter biocontrol ability and physiological state, and we must consider laboratory media in optimizing the use of biocontrol agents.

EFFECT OF HEAT TREATMENT ON NUTRITIONAL VALUE OF WINGED BEAN (Psophocarpus tetragonolobus) AS COMPARED TO SOYBEAN I. CHEMICAL CHARACTERISTICS OF TREATED WINGED BEAN

  • Mutia, R.;Uchida, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.1
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    • pp.19-26
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    • 1993
  • The effect heat treatment (autoclave) on nutritional value of winged bean as compared to soybean has been investigated. The winged bean and soybean were obtained from local cultivar grown in Indonesia. The beans were autoclaved at $120^{\circ}C$ for 15, 30, 45, 60 or 90 minutes, respectively before being ground for chemical analysis. Trypsin inhibitors of winged bean and soybean decreased (p < 0.05) along with decreasing of urease activity as heating time increased from 0 to 90 minutes. Heat treatment significantly (p < 0.05) reduced protein solubility in 0.2% potassium hydroxide of winged bean as well as soybean. In vitro protein digestibility was significantly (p < 0.05) improved by heating treatment (15 to 60 min of autoclaving), however, excessive heating (90 min of autoclaving) decreased the digestibility of winged beans. Excessive heating had adverse effect on lysine, cystine and methionine contents of winged beans. The results of this study suggested that autoclaving at $120^{\circ}C$ within 45 minutes should be adequate to remove protease inhibitors and could improve protein digestibility of winged beans.