• Title/Summary/Keyword: acid proteinase

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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.

Identification of a pr 1-like Gene of Entomopathogenic Fungus, Beauveria bassiana F-101 Isolated from Thecodiplosis japonensis

  • Shin Sang Chul;Roh Jong Yul;Shim Hee Jin;Kim Soon Kee;Kim Chul Su;Park Il Kwon;Jeon Mun Jang;Je Yeon Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.2
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    • pp.131-136
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    • 2005
  • Beauveria bassiana F-101, which has high toxicity toward Acantholyda parki as well as Thecodiplosis japonensis, was an isolate to develop an alternative control system against the major forest pests. Up to now, in B. bassiana, only one pr1 gene has been isolated and characterized. Therefore, we here reported the identification of a pr1-like gene, which would be a factor of toxicity from B. bassiana F-101. The oligonucleotides for the amplification of the pr1-like gene, were chosen based on the conserved regions of the subtilisin family enzymes, pr1 genes of B. bassiana and Metarhizium anisopliae, and proteinase K of Tritirachium album. The cloned PCR fragment had 1111 bp including 52 bp intron. The deduced Pr1-like peptide showed a low identity with Pr1s of entomopathogenic fungi such as B. bassiana Pr1 (BbPr1) and M. anisopliae Pr1 (MaPr1) as well as the proteinase K of T. album (TaPrK). Instead, the deduced peptide had a substantially high amino acid sequence identity $(>65\%)$ with the serine proteases of Magnaporthe grisea (MgSPM1) and Podospora anserina (PaPspA). These results, therefore, appear to suggest that the putative Pr1-like peptide of B. bassiana F-101 belongs to the subtilisin-like serine protease family and may be a novel gene.

Characterization of the Bacteriocin Produced by a Leuconostoc mesenteroides Strain Inhibiting the Growth of Lactobacillus sakei (Lactobacillus sakei 생육저해활성 보유 Leuconostoc mesenteroides가 생산하는 Bacteriocin의 특성)

  • Lee, Kwang-Hee;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.390-396
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    • 2011
  • Lactobacillus sakei is known to be the most populous lactic acid bacteria in over-ripened kimchi. Twenty three strains of Leuconostoc species inhibiting the growth of Lb. sakei were isolated from kimchi and amongst these the Leuconostoc mesenteroides strain CK0122 exhibited the highest antagonistic activity against Lb. sakei. The culture supernatant of the strain CK0122 was fractionated by a molecular weight cutter and lyophilized. The fraction with a molecular weight of less than 3,000 Da showed antagonistic activity against Lb. sakei. The antibacterial activity of the active fraction was sensitive to proteinase K treatment, confirming its proteinaceous nature (bacteriocin). The crude bacteriocin was active in the pH range of 4 to 7 and extremely stable after 15 min of heat treatment at $121^{\circ}C$. The crude bacteriocin inhibited the growth of Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Alcaligenes xylosoxydans, Flavobacterium sp., and Salmonella typhimurium.

Proteolytic Enzymes Distributed in the Tissues of Dark Fleshed Fish 1. Comparison of the Proteolytic Activity of the Tissue Extracts from the Meat of Mackerel and Sardine (혈합육어의 조직중에 분포하는 단백질분해효소 1. 고등어와 정어리 육조직중의 단백질분해효소의 활성비교)

  • PYEUN Jae-Hyeung;KIM Hyeung-Rak;CHO Jin-Guen
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.5
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    • pp.469-476
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    • 1986
  • Proteolytic activity of the tissue extracts from the muscle of mackerel, Scomber japonicus, and sardine, Sardinops melanostcta, was comparence with referenced to the optimum reaction condition. Thermal stability and change of proteolytic activity of the tissue extracts during storage were investigated. The existence of acid, weak acid and alkaline proteinase was identified in the ordinary and dark muscle of the mackerel and sardine. Specific activity of acid proteinase was stronger than weak acid or alkaline proteinase in the both fish. The proteolytic activity of the tissue extracts on the optimum reaction condition was: ordinary muscle of mackerel, 0.12 nM-Tyr. eq./mg-prot. /min. at pH 3.0 and $50^{\circ}C$; dark muscle of mackerel, 0.36 nM-Tyr. eq./mg-prot. /min. at pH 3.0 and $45^{\circ}C$; ordinary muscle of sardine, 0.45 nM-Tyr. eq./mg-prot. /min. at pH 2.4 and $45^{\circ}C$; dark muscle of sardine, 0.24 nM-Tyr. eq./mg-port. /min. at pH 2.4 and $45^{\circ}C$. The proteinases distributed in the muscle of mackerel and sardine were stable with the heat treatment at $45^{\circ}C$ for 5 minutes, but those in the dark muscle of mackerel was stable with the treatment at $5^{\circ}C$ for 5 minutes. The proteinases from the muscle were slowly inactivated with the whole storage days at $5^{\circ}C\;and\;-15^{\circ}C$, those were more stable at $-15^{\circ}C\;than\;5^{\circ}C$ storage.

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Purification and Characterization of Fibrinolytic Enzyme from Armillariella mellea (뽕나무버섯으로부터 Fibrinolytic enzyme의 정제 및 특성 연구)

  • Kim, Jun-Ho;Kim, Yang-Sun
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.583-588
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    • 1998
  • A fibrinolytic enzyme has been isolated from the edible honey mushroom, Armillariella mellea and purified. The apparent molecular mass of purified enzyme was estimated to be 19800Da by SDS polyacryl amide gel electrophoresis and 19900Da by gel filtration, indicating that it was a monomer. The enzyme was optimal at pH 7, suggesting that the purified enzyme was a neutral proteinase. It shows the maximum fibrinolytic activity at $55^{\circ}C$, is completely inactivated above $65^{\circ}C$, and still indicates 40% of activity at $37^{\circ}C$. The fibrinolytic activity has been decreased by the addition of EDTA. Fifteen amino acid sequence was determined by protein sequencing techniques.

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Immunostimulating Components from the Root of Angelica gigas Nakai (참당귀(Angelica gigas Nakai) 뿌리의 면역증강활성 성분)

  • Ahn, Kyung-Seop;Sim, Woong-Seop;Kim, Hwan-Mook;Han, Sang-Bae;Kim, Ik-Hwan
    • Korean Journal of Pharmacognosy
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    • v.27 no.3
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    • pp.254-261
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    • 1996
  • A potent immuno-stimulating activity was detected from the watersoluble and ethanol-precipitated crude extract (AG-1) of the root of Angelica gigas Nakai. The crude extract was fractionated into two fractions, an acidic AG-2 and a neutral AG-3 fraction by DEAE-cellulose adsorption. The two fractions contained polysaccharides of which M.W. were 10 Kdal and >2,000 Kdal respectively, proteins, and various inorganic components. The immunostimulating activities of two fractions were not reduced by proteinase K, acid or alkali treatment. The polysaccharides obtained from the root of A. gigas were mainly composed of arabinose, galactose, and galacturonic acid. These results indicated that immuno- stimulating components of the root of A. gigas was a kind of pectic polysaccharides or arabinogalactans.

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Thrombin Induced Apoptosis through Calcium-Mediated Activation of Cytosolic Phospholipase A2 in Intestinal Myofibroblasts

  • Mi Ja Park;Jong Hoon Won;Dae Kyong Kim
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.59-67
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    • 2023
  • Thrombin is a serine protease that participates in a variety of biological signaling through protease-activated receptors. Intestinal myofibroblasts play central roles in maintaining intestinal homeostasis. In this study, we found that thrombin-induced apoptosis is mediated by the calcium-mediated activation of cytosolic phospholipase A2 in the CCD-18Co cell. Thrombin reduced cell viability by inducing apoptosis and proteinase-activated receptor-1 antagonist attenuated thrombin-induced cell death. Endogenous ceramide did not affect the cell viability itself, but a ceramide-mediated pathway was involved in thrombin-induced cell death. Thrombin increased intracellular calcium levels and cytosolic phospholipase A2 activity. The ceramide synthase inhibitor Fumonisin B1, intracellular calcium chelator BAPTA-AM, and cytosolic phospholipase A2 inhibitor AACOCF3 inhibited thrombin-induced cell death. Thrombin stimulated arachidonic acid release and reactive oxygen species generation, which was blocked by AACOCF3, BAPTA-AM, and the antioxidant reagent Trolox. Taken together, thrombin triggered apoptosis through calcium-mediated activation of cytosolic phospholipase A2 in intestinal myofibroblasts.

ISOLATION AND CHERACTERIZATION OF ACTINIDIN GENE FROM CHINESE WILD KIWI FRUIT

  • Lee, Nam-Keun;Hahm, Young-Tae
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.527-530
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    • 2000
  • A kiwi fruit ,called as the Chinese gooseberry, is originated from the Yangtze River Valley of Northern China and Zhejiang Province on the cost of Eastern China. Around 1950, a large mass production began at New Zealand with an Improved breeding. Plant origin actinidin from kiwi fruit belongs to the papain family of cysteine proteinase, which in plants includes papain from papaya, bromelain from pineapple, Cl4 protease from tomato and aleurain from barley. Actinidin is involved in the ripening-related gene family. In this study, protease gene of chinese wild kiwi fruit was isolated and characterized. 1.2kb PCR-amplified fragment was obtained from the total RNA using RT-PCR. pWACT-1 was obtained by subcloning of amplified fragment into pGEM-T Easy cloning vector and analyzed nucleotide sequence by DNA sequencing and amino acid sequence. In Result, high levels of homology between wild kiwi and New Zealand cultured-kiwi was obtained.

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Optimization of Protein Extraction for Lichen Thalli

  • Kondratiuk, Anna S.;Savchuk, Oleksiy M.;Hur, Jae-Seoun
    • Mycobiology
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    • v.43 no.2
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    • pp.157-162
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    • 2015
  • Lichen-forming fungal proteins have been seldom searched due to many difficulties in their extraction. Phenols, quinones, proteases, and other components released during cell disruption have been known to be the greatest challenges related to protein extraction from lichens. To overcome these problems and maintain good electrophoretic resolution and high protein concentration, an extraction buffer containing polyvinylpolypyrrolidone, ascorbic acid, Triton X-100, polyethylene glycol, proteinase, and oxidase inhibitors in sodium phosphate buffer was developed. This extraction buffer showed high efficiency for all lichen species tested in the study.

Plant Growth Regulator Produced by Streptomyces sp. (Part II) Conditions of Production and Some Properties of the Plant Growth Regulator

  • 김광현;서정훈
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1978.10a
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    • pp.207.5-208
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    • 1978
  • Effects of the plant growth regulator (P. G. R.)on the reaction of proteinase, $\gamma-amylase$ and acid phosphatase were investigated, and also were the conditions of production of P. G. R. by Stroptomyces sp. 445. The P. G. R. had no effect on the act ivities of such enzymes in mung bean seedling. But in germinating seed previously treated with P. G. R. it effected the activity of protease in cotyledon. In the conditions of production of P. G. R., the maxim, activity was appeared in shaking cutlure at $30^{\circ}C$ for 5 days, and by the addition of peptone or casein hydrolysate as nitrogen source, soluble starch as carbon source, and sulfur as metal ion.

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