• Title/Summary/Keyword: proteinase K

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Biomolecular Strategies for Preparation of High Quality Surimi-Based Products

  • Nakamura Soichiro;Ogawa Masahiro
    • Preventive Nutrition and Food Science
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    • v.10 no.2
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    • pp.191-197
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    • 2005
  • There exist two interesting phenomena in making seafood products from surimi. When salted surimi is kept at a constant low temperature $(4\~40^{\circ}C)$, its rheological properties change from sol to gel, which is called 'setting'. Seafood processors can exploit changes that occur during setting in preparation of surimibased products, because heating at high temperatures, after the pre-heating during the setting process, enhances the gel-strength of salted surimi. Contrarily, when salted surimi or low-temperature set gel is heated at moderate temperatures $(50\~70^{\circ}C)$, a deterioration of gel is observed. The phenomenon is termed 'modori'. In the modori temperature range, heat-stable cysteine proteinases such as cathepsin B, H, Land L-Iike hydrolyze the myosins responsible for gel-formation, resulting in gel weakening modori. This article reviews molecular events occurring during gel setting that improve the quality of surimi-based products, and inhibition of modori by applying proteinase inhibitors. Application of recombinant protein technology to surimi-based products is introduced and its prospects for practical use are discussed.

Effect of Proteinase Activity on the Cheddar Cheese Quality (단백분해 효소 활성(蛋白分解 酵素 活性)이 Cheddar Cheese의 품질(品質)에 미치는 영향(影響))

  • Kim, Min-Bae
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.2
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    • pp.157-164
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    • 1996
  • This study aimed increase the quality during ripening of Cheddar cheese made with proteinase-negative mutant of Streptococcus lactis KCTC 1913 selected by curing. The degradation of protein during cheese ripening were investigated by electrophoresis and chromatography. The results were summarized as follow ; 1. The number of lactic acid bacteria decreased with the ripening stage, and that of the control cheese decreased faster than that of the cheese made with mutant. 2. Polyacrylamide gel electrophoretic analysis of cheese caseins revealed no difference between the cheese made with mutant and the control cheese, but differences along with the ripening stage were evident. 3. On Sephadex G-25 column chromatography, the extracts of bitter components from the green cheese and 3 month ripended cheese were fractionated into 3 fractions. With the progress of ripening, bitter peptides were degraded to rather small peptides or free amino acids. 4. Sensory evaluation of the 3 month ripended Cheddar cheese found no significant differences in color but the cheese made with mutant evidenced higher palatability in flavor and better texture than the control cheese. 5. The yields of the cheddar cheese made with mutant was 0.14% higher than that of the control cheese.

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Natural Products as Manipulators of Rumen Fermentation

  • Wallace, R. John;McEwan, Neil R.;McIntosh, Freda M.;Teferedegne, Belete;Newbold, C. James
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.10
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    • pp.1458-1468
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    • 2002
  • There is increasing interest in exploiting natural products as feed additives to solve problems in animal nutrition and livestock production. Essential oils and saponins are two types of plant secondary compounds that hold promise as natural feed additives for ruminants. This paper describes recent advances in research into these additives. The research has generally concentrated on protein metabolism. Dietary essential oils caused rates of NH$_3$ production from amino acids in ruminal fluid taken from sheep and cattle receiving the oils to decrease, yet proteinase and peptidase activities were unchanged. Hyper-ammonia-producing (HAP) bacteria were the most sensitive of ruminal bacteria to essential oils in pure culture. Essential oils also slowed colonisation and digestion of some feedstuffs. Ruminobacter amylophilus may be a key organism in mediating these effects. Saponin-containing plants and their extracts appear to be useful as a means of suppressing the bacteriolytic activity of rumen ciliate protozoa and thereby enhancing total microbial protein flow from the rumen. The effects of some saponins seems to be transient, which may stem from the hydrolysis of saponins to their corresponding sapogenin aglycones, which are much less toxic to protozoa. Saponins also have selective antibacterial effects which may prove useful in, for example, controlling starch digestion. These studies illustrate that plant secondary compounds, of which essential oils and saponins comprise a small proportion, have great potential as 'natural' manipulators of rumen fermentation, to the potential benefit of the farmer and the environment.

Bacillus subtilis as a Tool for Screening Soil Metagenomic Libraries for Antimicrobial Activities

  • Biver, Sophie;Steels, Sebastien;Portetelle, Daniel;Vandenbol, Micheline
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.850-855
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    • 2013
  • Finding new antimicrobial activities by functional metagenomics has been shown to depend on the heterologous host used to express the foreign DNA. Therefore, efforts are devoted to developing new tools for constructing metagenomic libraries in shuttle vectors replicatable in phylogenetically distinct hosts. Here we evaluated the use of the Escherichia coli-Bacillus subtilis shuttle vector pHT01 to construct a forest-soil metagenomic library. This library was screened in both hosts for antimicrobial activities against four opportunistic bacteria: Proteus vulgaris, Bacillus cereus, Staphylococcus epidermidis, and Micrococcus luteus. A new antibacterial activity against B. cereus was found upon screening in B. subtilis. The new antimicrobial agent, sensitive to proteinase K, was not active when the corresponding DNA fragment was expressed in E. coli. Our results validate the use of pHT01 as a shuttle vector and B. subtilis as a host to isolate new activities by functional metagenomics.

Isolation of Bacteriocin-producing Lactic Acid Bacteria from Human Intestines and the Characteristics of their Bacteriocins (Bacteriocin을 생산하는 장내 유산균의 분리 및 Bacteriocin 특성조사)

  • 김정환;맹길재;김정상;지근억
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1228-1236
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    • 1997
  • Lactobacillus strains were isolated from volunteer's feces, including from newly-born infants and adults in their 20's, by using differential MRS-BPB plates. Total 56 presumptive Lactobacillus strains were isolated and the bacteriocin productions by the isolates were examined by agar diffusion method. Six bacteriocin-producing strains were confirmed. Among them, two isolates, HU-1 and H22-3, showed the most outstanding antimicrobial activities, which were not affected by pH adjustments or catalase treatments of culture. HU-1 was originated from a two-years old boy and H22-3 was originated from a newly-born infant. HU-1 and H22-3 had the same morphology(short rod) when examined by scanning electron microscope, and the same biochemical traits including growth temperature range, salt tolerance and sugar-fermenting abilities. But the growth-inhibition spectrum and plasmid profiles of HU-1 and H22-3 were different. Both strains inhibited the growth of various Gram (+) microorganisms including Listeria monocytogenes. Micrococcus luteus, and Staphylococcus aureus in addition to many species of lactic acid bacteria, indicating the production of broad-spectrum bacteriocins. Bacteriocins produced by HU-1 and H22-3 were stable up to 90℃, 15 min heat treatments. Their activities were not affected by pepsin or trypsin treatments but destroyed by proteinaseK or pronase treatments.

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Anticariogenic Activities of Lactobacillus sakei K-7 Isolated from Kimchi (김치로부터 분리한 Lactobacillus sakei K-7의 항충치 활성 특성)

  • Moon, Jin-Seok;Ahn, Ji-Eun;Han, A-Reum;Heo, Jeong-Seon;Eom, Hyun-Ju;Shin, Chul-Soo;Choi, Hye-Sun;Han, Nam-Soo
    • KSBB Journal
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    • v.26 no.6
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    • pp.513-516
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    • 2011
  • The occurrence of dental caries is mainly associated with oral pathogens, especially cariogenic Streptococcus mutans. The aim of this study was to isolate and characterize lactic acid bacterium showing inhibitory activity against cariogenic Streptococcus mutans. As results, an isolate with strong inhibitory activity was obtained from Kimchi and it was identified as Lactobacillus sakei by API and 16S rRNA gene analyses. This strain secreted an inhibitory compound in cell growth medium and the activity of the compound was completely disappeared by proteinase K revealing the fact that the compound is proteinous substance, bacteriocin. Optimal culture condition for bacteriocin production by Lb. sakei K-7 was at pH 7.5 and $37^{\circ}C$ for 18 h. Oral administration of this isolate may give anticariogenic and probiotic effects on hosts.

Properties of a Bacteriocin Produced by Bacillus subtilis EMD4 Isolated from Ganjang (Soy Sauce)

  • Liu, Xiaoming;Lee, Jae Yong;Jeong, Seon-Ju;Cho, Kye Man;Kim, Gyoung Min;Shin, Jung-Hye;Kim, Jong-Sang;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1493-1501
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    • 2015
  • A Bacillus species, EMD4, with strong antibacterial activity was isolated from ganjang (soy sauce) and identified as B. subtilis. B. subtilis EMD4 strongly inhibited the growth of B. cereus ATCC14579 and B. thuringiensis ATCC33679. The antibacterial activity was stable at pH 3-9 but inactive at pH 10 and above. The activity was fully retained after 15 min at 80℃ but reduced by 50% after 15 min at 90℃. The activity was completely destroyed by proteinase K and protease treatment, indicating its proteinaceous nature. The bacteriocin (BacEMD4) was partially purified from culture supernatant by ammonium sulfate precipitation, and Q-Sepharose and Sephadex G-50 column chromatographies. The specific activity was increased from 769.2 AU/mg protein to 8,347.8 AU/mg protein and the final yield was 12.6%. The size of BacEMD4 was determined to be 3.5 kDa by Tricine SDS-PAGE. The N-terminal amino acid sequence was similar with that of Subtilosin A. Nucleotide sequencing of the cloned gene confirmed that BacEMD4 was Subtilosin A. BacEMD4 showed bactericidal activity against B. cereus ATCC14579.

Overexpression of jasmonic acid carboxyl methyltransferase increases tuber yield and size in transgenic potato

  • Sohn, Hwang-Bae;Lee, Han-Yong;Seo, Ju-Seok;Jung, Choon-Kyun;Jeon, Jae-Heung;Kim, Jeong-Han;Lee, Yin-Won;Lee, Jong-Seob;Cheong, Jong-Joo;Choi, Yang-Do
    • Plant Biotechnology Reports
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    • v.5 no.1
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    • pp.27-34
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    • 2011
  • Jasmonates control diverse plant developmental processes, such as seed germination, flower, fruit and seed development, senescence and tuberization in potato. To understand the role of methyl jasmonate (MeJA) in potato tuberization, the Arabidopsis JMT gene encoding jasmonic acid carboxyl methyltransferase was constitutively overexpressed in transgenic potato plants. Increases in tuber yield and size as well as in vitro tuberization frequency were observed in transgenic plants. These were correlated with JMT mRNA level-- the higher expression level, the higher the tuber yield and size. The levels of jasmonic acid (JA), MeJA and tuberonic acid (TA) were also higher than those in control plants. Transgenic plants also exhibited higher expression of jasmonate-responsive genes such as those for allene oxide cyclase (AOC) and proteinase inhibitor II (PINII). These results indicate that JMT overexpression induces jasmonate biosynthesis genes and thus JA and TA pools in transgenic potatoes. This results in enhanced tuber yield and size in transgenic potato plants.

An Immunological Approach to ABA Receptor and its Gene

  • Xie Zhou;Jin, Zhen-hua;Zheng, Zhi-fu;Kai Xia;Zhang, Neng-gang;Wan, Yin-sheng;Sang, Yong-ming;Chen, Kao-shan;Liu, Shi-ming
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.06a
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    • pp.68-78
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    • 1996
  • Two types of immunoloigcal probes, anti-ABBP Abs, have been developed. The purified ABBP from ABA-C1-BSA-sepharose 4B column was identified by PAGE and appeared in one band of about 56KD, as well as showed a specific binding ability and a high affinity for ABA (Kd2.0$\times$10-9 mol/L). Unexpectedly, the existence of rRNA with a length of around 300 nucleotides could be found, when the ABBP was digested with proteinase K and identified by eletrophorsis on an agarose gel (1%). As a result, about 120 cDNA clones coding maize 17s RNA and only one cDNA clone coding ABBP (24cDNA) were obtained from 200,000 seperated phage plaques by the anti-ABBP pAbs. 24cDNA had 1075bp and contained an open reading frame coding 254 amino acids. The anti-idiotypic Ab raised against an ABA MAb showed the ability of either mimicking ABA or competing with ABA. The localization of ABBPs in plant cell was investigated.

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First Evidence that Sika Deer (Cervus nippon) Velvet Antler Extract Suppresses Migration of Human Prostate Cancer Cells

  • Tang, YuJiao;Jeon, Byong-Tae;Wang, Yanmei;Choi, Eun-Ju;Kim, Yon-Suk;Hwang, Jin-Woo;Park, Pyo-Jam;Moon, Sang Ho;Kim, Eun-Kyung
    • Food Science of Animal Resources
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    • v.35 no.4
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    • pp.507-514
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    • 2015
  • Deer velvet antler (DVA) is one of the most popular medicines in China. Numerous studies have demonstrated that velvet antler possess biological effects. However, data regarding its anti-migration activity on prostate cancer is scarce. In this study, we investigated the inhibitory effect of top DVA (T-DVA) on the expression of prostate-specific antigen (PSA) and migration-related genes in the human prostate cancer cell, LNCaP. The T-DVA down-regulated the expression of PSA. In addition, the RadiusTM assay revealed that T-DVA inhibited the migration behavior of prostate cancer cells. Furthermore, the expression of matrix metalloproteinase (MMP)-9 and vascular endothelial growth factor (VEGF) was also decreased with T-DVA. On the contrary, T-DVA increased the tissue inhibition of metallo-proteinase (TIMP)-1 and (TIMP)-2. Taken together, our findings indicate that the T-DVA possesses anti-migration activity on prostate cancer cells. This is the first study of DVA to report the anti-migration activity on prostate cancer.