• Title/Summary/Keyword: Proteinase

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Fermentation of rice flour with Weissella koreensis HO20 and Weissella kimchii HO22 isolated from kimchi and its use in the making of jeolpyeon (김치유산균(Weissella koreensis HO20, Weissella kimchii HO22)으로 발효한 쌀가루의 이화학적 특성 및 이를 이용한 절편의 제조)

  • Choi, Hyejung;Lee, Hwawon;Yoon, Sun
    • Korean journal of food and cookery science
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    • v.29 no.3
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    • pp.267-274
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    • 2013
  • Demand for a rice cake, a popular traditional food in Korea, is rising, but its industrial-scale production is extremely difficult due to its short shelf-life caused by starch retrogradation and microbial spoilage. By means of the sourdough fermentation technique, we attempt to develop rice cakes with a longer shelf-life. Heterofermentative lactic acid bacteria (Weissella koreensis HO20, Weissella kimchii HO22) isolated from kimchi were used to ferment wet-milled rice flour for their abilities to produce exopolysaccharides and to inhibit the microbial spoilage of rice cakes. After 24 hr of fermentation at $25^{\circ}C$, viable cell counts in rice dough increased from $10^6$ CFU/g to $10^8$ CFU/g and total titratable acidity increased from 0.05% to 0.20%, whereas pH decreased from 6.5 to 5.1. Fermented rice flour showed significantly lower peak, trough, and final viscosities as well as breakdown and setback viscosities measured by rapid viscoanalyzer. Both lactic acid bacteria showed in vitro antifungal activity against Penicillium crustosum isolated from rice cakes. The antifungal activity remained constant after the treatments with heat, proteinase K and trypsin, but fell significantly by increase of pH. Rice cakes made of fermented rice flour were found to retard mycelial growth of P. crustosum. The degree of retrogradation as measured by the hardness of the rice cake was significantly reduced by the use of fermented rice flour. The results suggest that use of fermented rice flour has a beneficial role in retarding starch retrogradation and in preventing fungal growth, hence extending the shelf-life of rice cakes.

Profiling of differential expressed proteins from various explants in Platycodon grandiflorum

  • Kim, Hye-Rim;Kwon, Soo Jeong;Roy, Swapan Kumar;Kamal, Abu Hena Mostafa;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Cho, Kab Yeon;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.131-131
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    • 2017
  • Though the Platycodon grandiflorum, has a broad range of pharmacologic properties, but the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two-dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}2-fold$) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, the frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). Taken together, the protein profile may provide insight clues for better understanding the characteristics of proteins and its metabolic activities in various explants of this essential medicinal plant P. grandiflorum.

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Inhibition of Growth and Microcystin Toxicity, and Characterization of Algicidal Substances from Lactobacillus graminis against Microcystis aeruginosa (Microcystis aeruginosa에 대한 Lactobacillus graminis의 성장 억제능, microcystin 분해 및 살조 물질의 특성)

  • Joo, Jae-Hyoung;Park, Bum Soo;Lee, Eun-Seon;Kang, Yoon-Ho;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
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    • v.49 no.3
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    • pp.176-186
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    • 2016
  • For several decades, lactic acid bacterium (Lactobacillus graminis: LAB) has been generally recognized as safe. To develop the pan-environmental bio-control agent, algicidal activity of the live LAB cell and its culture filtrate (CF) was examined against Microcystis aeruginosa. LAB cells perfectly lysed M. aeruginosa within 3 days, while the CF had a less effect than the live cells, approximately 78% inhibition of algal growth during a same culture period. The concentration of microcystin in alone culture of M. aeruginosa was $7.1{\mu}gL^{-1}$, but gradually increased and leach $158.5{\mu}gL^{-1}$ on 10 days. However, LAB cells clearly decreased the microcystin by $10.3{\mu}gL^{-1}$ in the same period, approximately 93.5%. CF of LAB showed a strong algicidal activity over 75% between pH 2-7, 91.3% by the treatment of proteinase K, 87.8% by below 3 kDa in particle size, and 75.3% by heat treatment, respectively. Of five solvents, fractions of CF passed through solvents diethyl ether and ethyl acetate showed an obvious algicidal activity in the algal-lawn test. Among 5 fractions purified by silica-gel TLC plate, two spots showed a most strong removal activity on M. aeruginosa. Another analysis of GC indicate that CF contained six representative fatty acids. Even though most of these substance have been known as an anti-algal substance against M. aeruginosa, oleic acid is the most effective. These results suggested that the culture filtrate or specific substances, like a fatty acids, in comparison with live L. graminis can be a successful and eco-friendly agent to control Microcystis bloom.

사람 난포액에 존재하는 Matrix Metalloproteinase-2 Isoform의 동정

  • 나경아;김지수;심명선;권혁찬;이승재;윤용달;김해권
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.108-108
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    • 2002
  • 포유동물의 암컷 생식기관에 존재하는 다양한 종류의 matrix metallo-proteinase (MMP)는 난소와 자궁의 구성성분의 주기적인 변화를 조절하며 이중 난소의 MMP는 난포의 성장과 배란 그리고 퇴화 동안 조직재구성에 매우 중요한 역할을 한다고 알려져 있다. 본 실험에서는 근래에 새로 발견된 사람의 난포액에 존재하는 분자량 약 110kDa인 MMP-2 isoform GA110을 동정하고 자 하였다. 난포액으로부터 GA110 단백질을 분리하기 위하여 난포액에 5mM ethylenediaminetetraacetic acid(EDTA)를 처리한 후 DEAE Sepharose Fast Flow를 이용한 chromatography를 시행하였다. 그 결과, 난포액 단백질들은 0.2M NaCl 의 분획에서 GA110 활성을 나타내었고 anti-human MMP-2 antibody에 대한 면역반응도 뚜렷이 나타났다. DEAE Sepharose Fast Flow에서 얻은 분획 중 GA110의 활성과 면역반응을 모두 나타내는 분획만을 모아 Gelatin Sepharose 4B affinity chromatography로 다시 분리하였다 분리한 결과 resin에 흡착된 단백질 (eluate) 분획들에서 매우 뚜렷한 GA110 gelatinase 활성을 나타내었으며 면역반응 또한 관찰되었다. 이 분획들의 단백질을 농축한 후 zymography를 시행하여 나타난 GA110 단백질 band를 잘라 내었으며 이로부터 단백질을 electroelution하여 농축한 후 reducing agent인 2-mercaptoethanol를 처리하였다. 이를 전기영동 후 MMP-2 (propeptide region) antibody를 사용하여 immunoblotting 한 결과 70-72kDa의 단백질만이 면역반응을 나타내었다. 마지막으로 위와 같이 준비된 70-72kDa 단백질의 아미노산 서열을 Edman degradation 방법으로 분석하였다. 그 결과 이 단백질의 N 말단의 10개의 아미노산 배열 순서가 알려진 사람의 proMMP-2의 전체 배열순서 중 propetide domain의 N 말단에서부터 다섯 번째에서 시작하여 10개의 아미노산의 서열과 정확하게 일치하였다. 위 결과들로 미루어 사람의 난포액에 존재하는 MMP-2의 새로운 isoform인 GA110은 70-72kDa의 ProMMP-2가 disulfide bond를 통해 homodimer 구조를 이루고 있는 것으로 여겨진다.

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Immunoblot patterns of clonorchiasis (면역이적법에 의한 간흡충 항원분획과 감염자의 항체반응 양상)

  • 홍성태;고원규
    • Parasites, Hosts and Diseases
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    • v.35 no.2
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    • pp.87-94
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    • 1997
  • Clonorchis sinearis is a liver fluke which is the most prevalent helminth of humans in Korea. The better diagnostic measure of clonorchiasis is required for its nationwide control program. The present study observed antigenic bands of C. sinensis and reacting immunoglobulins in serum of infected residents. Adult C. sinensis were recovered from experimentally infected rabbits and soluble crude extract of the worms was used as the antigen after supplementation of E-64, a cysteine proteinase inhibitor. SDS- PAGE of the crude antigen resolved more than 20 protein bands between 200 and 14 kDa. The sera of infected humans collected at an endemic village showed specific IgG and IgE antibodies but little IgM and IgA antibodies. The protein bands of 94, 80, 72, 68, 52, 47, 43, 37, 34, and 28-25 kDa strongly reacted with serum Ig(GMA) or IgG antibody and 64, 62, 52, 47,44, 34,28, and 26 kDa bands reacted with serum IgE antibody. However, the 94, 80, 72, 68, 64, 62, 52, 47, and 40 kDa bands of C. sinensis antigen were found non specific. The protein bands of 43, 34, and 28-25 kDa of C. sinensis are primary target molecules of further analysis.

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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.

Characterization of Two Algal Lytic Bacteria Associated with Management of the Cyanobacterium Anabaena flos-aquae

  • Kim, Jeong-Dong;Lee, Choul-Gyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.382-390
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    • 2006
  • Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

Immunohistochemical Study on the Effects of Cortex Acanthopanacis Senticosi on the Expression of Calpain in Collagen-Induced Arthritis in Rats (교원질로 유발된 관절염에서 가시오가피가 calpain의 발현에 미치는 영향에 대한 면역조직화학적 연구)

  • 오충환;김순중;서일복
    • The Journal of Korean Medicine
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    • v.23 no.4
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    • pp.98-104
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    • 2002
  • Objectives: Calpain, a calcium-dependent cysteine proteinase, may be one of the proteolytic enzymes that mediate cartilage degradation associated with rheumatoid arthritis. The object of this study is to ascertain immunohistochemically whether calpain is present in the inflamed joints of collagen-induced arthritis of rats, and examine the effect of Cortex Acanthopanacis Senticosi on the expression of calpain. Methods: Male Lewis rats, around 200g of body weight, were immunized with bovine type II collagen. After 3 weeks from first immunization, rats were divided into arthritic control (n=6) group and Cortex Acanthopanacis Senticosi-treated (n=6) group. Non-immunized rats served as the normal (n=6) group. All animals were sacrificed at 15 days post-treatment and tibiotarsal joints were removed. Calpain immunohistochemistry was performed on the midsagittal section of the tibiotarsal joint. Results: All animals of the control and treated groups showed ankylosing osteoarthritis. However, the animals of the treated group showed alleviation in the fibrous ankylosis, destruction of articular cartilage and destruction of subchondral bony tissue compared with the animals of the control group. Calpain was expressed in the chondrocyte lacunae of growing articular cartilage, in the skeletal muscle fibers, in the peripheral nerves, and in the vessel walls around the joints of all groups. In the control and treated groups, calpain was also expressed in proliferating synovial epithelia, subsynovial stroma cells, surface of articular cartilage, and fibrous pannus around destructive subchondral bony tissue. However, the expression density of calpain in the treated group was diminished compared with the control group, especially in surface of articular cartilage and fibrous pannus. Conclusions: These observations indicated that calpain plays an important role in the destruction of cartilage and bone in collagen-induced arthritis of rats, and also indicated that Cortex Acanthopanacis Senticosi inhibits the development of arthritis by decreasing the expression of calpain.

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Proteolytic System of Streptococcus thermophilus

  • Rodriguez-Serrano, G.M.;Garcia-Garibay, M.;Cruz-Guerrero, A.E.;Gomez-Ruiz, L.;Ayala-Nino, A.;Castaneda-Ovando, A.;Gonzalez-Olivares, L.G.
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1581-1588
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    • 2018
  • The growth of lactic acid bacteria (LAB) generates a high number of metabolites related to aromas and flavors in fermented dairy foods. These microbial proteases are involved in protein hydrolysis that produces necessary peptides for their growth and releases different molecules of interest, like bioactive peptides, during their activity. Each genus in particular has its own proteolytic system to hydrolyze the necessary proteins to meet its requirements. This review aims to highlight the differences between the proteolytic systems of Streptococcus thermophilus and other lactic acid bacteria (Lactococcus and Lactobacillus) since they are microorganisms that are frequently used in combination with other LAB in the elaboration of fermented dairy products. Based on genetic studies and in vitro and in vivo tests, the proteolytic system of Streptococcus thermophilus has been divided into three parts: 1) a serine proteinase linked to the cellular wall that is activated in the absence of glutamine and methionine; 2) the transport of peptides and oligopeptides, which are integrated in both the Dpp system and the Ami system, respectively; according to this, it is worth mentioning that the Ami system is able to transport peptides with up to 23 amino acids while the Opp system of Lactococcus or Lactobacillus transports chains with less than 13 amino acids; and finally, 3) peptide hydrolysis by intracellular peptidases, including a group of three exclusive of S. thermophilus capable of releasing either aromatic amino acids or peptides with aromatic amino acids.

Proteome Profiling Unfurl Differential Expressed Proteins from Various Explants in Platycodon Grandiflorum

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Cho, Kab-Yeon;Boo, Hee-Ock;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.97-106
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    • 2015
  • Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}$ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.