• Title/Summary/Keyword: Protease digestion

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Organic Acid Composition and Flavor Characteristics of Lactic Acid Fermented Cereal Beverages

  • Yi, Do-Youn;Kim, Gi-Myung;Lee, Ki-Young
    • Journal of Microbiology and Biotechnology
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    • v.3 no.2
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    • pp.129-133
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    • 1993
  • The effect of different compositions of organic acids on the flavor profile of 10% sugar solution was investigated by the response surface methodology, and the results were used to evaluate the flavor characteristics of lactic acid fermented cereal beverages. A mixture of extruded rice flour (10%) and soymilk (7.8% dry matter) was fermented with Leuconostoc mesenteroides (Sikhae). Depending on the substrate pretreatments, for example, the malt or amylase digestion and the proteolytic enzyme hydrolysis, the sugar and organic acid composition of the product varied. The organic acid composition of the fermented beverages was in the ranges of 0.44-0.55% lactic acid, 0.05-0.09% acetic acid and 0.07-0.09% citric acid, while that of commercial apple juice was 1.59% malic acid and 0.49% acetic acid. The flavor profiles of fermented beverages added with 10% sucrose were compared to those of apple juice and a model mixture containing 0.48% citric acid, 0.39% lactic acid and 0.12% acetic acid in 10% sugar solution. The QDA diagram of fermented beverages approached to that of apple juice, when the substrate was digested by amylase but not by protease.

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Functional Identification of an 8-Oxoguanine Specific Endonuclease from Thermotoga maritima

  • Im, Eun-Kyoung;Hong, Chang-Hyung;Back, Jung-Ho;Han, Ye-Sun;Chung, Ji-Hyung
    • BMB Reports
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    • v.38 no.6
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    • pp.676-682
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    • 2005
  • To date, no 8-oxoguanine-specific endonuclease-coding gene has been identified in Thermotoga maritima of the order Thermotogales, although its entire genome has been deciphered. However, the hypothetical protein Tm1821 from T. maritima, has a helix-hairpin-helix motif that is considered to be important for DNA binding and catalytic activity. Here, Tm1821 was overexpressed in Escherichia coli and purified using Ni-NTA affinity chromatography, protease digestion, and gel filtration. Tm1821 protein was found to efficiently cleave an oligonucleotide duplex containing 8-oxoguanine, but Tm1821 had little effect on other substrates containing modified bases. Moreover, Tm1821 strongly preferred DNA duplexes containing an 8-oxoguanine:C pair among oligonucleotide duplexes containing 8-oxoguanine paired with four different bases (A, C, G, or T). Furthermore, Tm1821 showed AP lyase activity and Schiff base formation with 8-oxoguanine in the presence of $NaBH_4$, which suggests that it is a bifunctional DNA glycosylase. Tm1821 protein shares unique conserved amino acids and substrate specificity with an 8-oxoguanine DNA glycosylase from the hyperthermophilic archaeon. Thus, the DNA recognition and catalytic mechanisms of Tm1821 protein are likely to be similar to archaeal repair protein, although T. maritima is an eubacterium.

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.

Identificaiton of the dITP- and XTP-Hydrolyzing Protein from Escherichia coli

  • Chung, Ji-Hyung;Park, Hyun-Young;Lee, Jong-Ho;Jang, Yang-Soo
    • BMB Reports
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    • v.35 no.4
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    • pp.403-408
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    • 2002
  • A hypothetical 21.0 kDa protein (ORF O197) from Escherichia coli K-12 was cloned, purified, and characterized. The protein sequence of ORF O197(termed EcO197) shares a 33.5% identity with that of a novel NTPase from Methanococcus jannaschii. The EcO197 protein was purified using Ni-NTA affinity chromatography, protease digestion, and gel filtration column. It hydrolyzed nucleoside triphosphates with an O6 atom-containing purine base to nucleoside monophosphate and pyrophosphate. The EcO197 protein had a strong preference for deoxyinosine triphosphate (dITP) and xanthosine triphosphate (XTP), while it had little activity in the standard nucleoside triphosphates (dATP, dCTP, dGTP, and dTTP). These aberrant nucleotides can be produced by oxidative deamination from purine nucleotides in cells; they are potentially mutagenic. The mutation protection mechanisms are caused by the incorporation into DNA of unwelcome nucleotides that are formed spontaneously. The EcO197 protein may function to eliminate specifically damaged purine nucleotide that contains the 6-keto group. This protein appears to be the first eubacterial dITP-and XTP-hydrolyzing enzyme that has been identified.

Seed Transmission Rates of Bean pod mottle virus and Soybean mosaic virus in Soybean May Be Affected by Mixed Infection or Expression of the Kunitz Trypsin Inhibitor

  • Nam, Moon;Bae, Hanhong;Hammond, John;Domier, Leslie L.;Youn, Young-Nam;Lee, Bong-Choon;Lim, Hyoun-Sub
    • Research in Plant Disease
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    • v.19 no.2
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    • pp.114-117
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    • 2013
  • To facilitate their spread, plant viruses have developed several methods for dispersal including insect and seed transmission. While insect transmission requires virus stability against insect digestion, seed-transmitted viruses have to overcome barriers to entry into embryos. Bean pod mottle virus (BPMV) is transmitted through seed at levels typically below 0.1%, but co-infection with Soybean mosaic virus (SMV) enhanced the seed transmission rate of BPMV in one experiment. In contrast, the rate of SMV seed transmission was not affected by BPMV co-infection. In a second preliminary study, the rate of SMV transmission was lower in an isoline of Williams 82 that contained a null mutation for the Kunitz trypsin inhibitor gene than in Williams 82. In this preliminary study, we observed that factors such as protease inhibitor expression and dual infection may affect the frequency of seed transmission of BPMV and SMV.

Rapid Isolation of Genomic DNA from Normal and Apoptotic Cells Using Magnetic Silica Resins

  • Park, Jee-Sun;Park, Jung-Hyun;Na, Shin-Young;Choe, Soo-Young;Choi, Sang-Nam;You, Kwan-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.890-894
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    • 2001
  • The isolation of genomic DNA from mammalian cells is usually performed by cell lysis followed by protein digestion, extraction, and finally, ethanol precipitation of the chromosomal DNA. However, in the case of large sample numbers or when only small amounts of starting materials are available, such conventional methods are not efficient and are cumbersome to be applied. Some alternative methods have been described as well as having commercial DNA isolation kits to be available, nevertheless, there is room left for much improvement. In the present study, a novel method is introduced, where it simplifies conventional protocols by omitting some time-consuming steps such as protease incubation or DNA precipitation and its resuspension. Using paramagnetic silica resins, the genomic DNA was purified over a magnetic field, and the bound DNA was eluted with a low-salt buffer. The fidelity and effectiveness of this novel method was determined by using normal and apoptotic cells as a starting material and then compared to other protocols. The high speed and convenience along with its high efficiency in detecting apoptotic chromosomal DNA will prove this method to be an improved alternative in the isolation of genomic DNA from mammalian cells.

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Characterization of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju

  • Lim, Sung-Mee;Park, Mi-Yeon;Chang, Dong-Suck
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.49-57
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    • 2005
  • Enterococcus faecium MJ-14, having strong antilisterial activity, was isolated from Korean fermented food, Meju. MJ-14 showed the same phenotypic characteristics, but different sugar utilization, as reference strain, E. faecium KCCM12118. It could utilize D-xylose, amygdaline, and gluconate, whereas E. faecium KCCM12118 could not. Optimal condition for bacteriocin production by E. faecium MJ-14 was at $37^{\circ}C$ and pH 7.0. Bacteriocin activity appeared in mid exponential phase and increased rapidly up to stationary phase. Activity was significantly promoted in MRS broth containing 3.0% glucose, 1.5% lactose, 2.0% peptone, or 1.5% tryptone. Bacteriocins effectively inhibited Enterococcus faecalis and Listeria spp. of Gram-positive bacteria, and Helicobacter pylori of Gram-negative bacteria, but did not inhibit yeasts and molds. They were stable against heat (for 30 min at $100^{\circ}C$), pH (3.0-9.0), long-term storage (for 60 days at 4 or $-20^{\circ}C$), and enzymatic digestion by catalase, proteinase K, papain, lysozyme, trypsin, chymotrypsin, and lipase, etc. Bacteriocin activity was completely inhibited by protease and pepsin, and 50% by ${\alpha}$-amylase. Studies on PCR detection of enterocin structural genes revealed bacteriocins are identical to enterocins A and B.

Preparation of Egg White Liquid Hydrolysate (ELH) and Its Radical-Scavenging Activity

  • Noh, Dong Ouk;Suh, Hyung Joo
    • Preventive Nutrition and Food Science
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    • v.20 no.3
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    • pp.183-189
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    • 2015
  • In the present study, an optimum protease was selected to hydrolyze the egg white liquid protein for the antioxidant peptides. Alcalase treatment yielded the highest amount of ${\alpha}$-amino groups (15.27 mg/mL), while the control (no enzymatic hydrolysis) showed the lowest amount of ${\alpha}$-amino groups (1.53 mg/mL). Alcalase also gave the highest degree of hydrolysis (DH) value (43.2%) and was more efficient for egg white liquid hydrolysis than the other enzymes. The Alcalase hydrolysate had the highest radical-scavenging activity (82.5%) at a concentration of 5.0 mg/mL. The conditions for enzymatic hydrolysis of egg white liquid with Alcalase were selected as substrate : water ratio of 2:1. Five percent Alacalse treatment did not show significant (P>0.05) increases of DH and ${\alpha}$-amino nitrogen content after 24 hhydrolysis. Thirty two hour-hydrolysis with 5% Alcalase is sufficient to make antioxidative egg white liquid hydrolysate from egg white liquid. DPPH and ABTS radical-scavenging activities were significantly (P<0.05) higher after enzymatic digestion. These results suggest that active peptides released from egg-white protein are effective radical-scavengers. Thus, this approach may be useful for the preparation of potent antioxidant products.

Sludge reduction by Enzyme Pretreatment (효소 전처리를 통한 슬러지 저감)

  • 김정래;심상준;최수형;염익태
    • KSBB Journal
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    • v.19 no.2
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    • pp.93-97
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    • 2004
  • We investigate the effect of enzyme pretreatment using protease, carbohydrase, and lipase on improvement of sludge treatment efficiency by measuring SCOD and TCOD. The enzyme-pretreatment increases SCOD of excess sludge. In addition, the amount of sludge reduction during digestion, in terms of SCOD and TCOD, are enhanced by enzyme-pretreatment. Among pretense, carbohydrase, and lipase, pretense showed the best enhancement of the sludge treatment efficiency. Sludge digestion followed by ozone and enzyme treatments showed more effective sludge treatment when compared with ozone treatment alone. Therefore, we expect that enzyme pretreatment can be used as a useful tool for enhancing the sludge treatment efficiency.

Preparation of Calcium-fortified Soymilk and in Vitro Digestion Properties of Its Protein and Calcium (칼슘강화 두유의 제조 및 단백질과 칼슘의 체외 소화특성)

  • Pyun, Jin-Won;Hwang, In-Kyeong
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.995-1000
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    • 1996
  • The present study was attempted to preprare calcium-fortified soymilk using proteases to improve calcium intolerance of soymilk protein and to evaluate its nutritional properties. The protease from Bacillus polymyxa was chosen as an enzyme source because it produced the least bitter taste and calcium-aggregation of soymilk among various enzymes. The optimum treatment time was 10 minutes at $50^{\circ}C$ for the best result. In vitro protein digestibility of calcium-fortified soymilks was comparable with that of control soymilk. Calcium in the digested soymilks was mostly in the ionic form and the amount of ionic calcium increased in accordance with the amount of fortified calcium in soymilk. This suggests that fortified calcium in the soymilk is bioavailable.

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