• Title/Summary/Keyword: 항세균성 단백질

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Purification of Antibacterial Proteins in Heliothis assulta Injected with Bacillus thuringiensis (Bacillus thurinRiens영의 주입에 의해 유도된 담배나방의 항세균 단백질의 정제)

  • 유종명;정성은이형철
    • The Korean Journal of Zoology
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    • v.37 no.2
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    • pp.274-280
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    • 1994
  • 담배나방(Heliothis assulta) 혈림프 항세균성 단백질의 특성을 알아보기 위해 종령 유충의 혈강으로 고라구연us thuringiens논 균을 주입하였다. 혈림프의 항세근적 활성은 주입후 48시간 경과군에서 가장 높았으며, E. coli와 풍 thuringiensis에 대하여 항세균성을 보이는 두 개의 단백질(G-1 및 G-11)들을 cation exchange chromatography와 gel filtration을 통해 정제하였다 49KD와 46KB의 두 subunit로 구성된 G-1은 높은 glvcine(17 13 mole %) 및 alanine(13.88 mole %) 함량을 보였으며, 14KD의 단일 pclvpeptide인 G-11에서는 histidine(16 14 mole %)과 glvclne(14.27 mole %)의 함량이 높아, 이러한 높은 glvcine 함량에 있어서 곤충에서 나타나는 다른 항세균성 단백질과 공통점을 보인다.

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Characterization and Expression of Antibacterial Protein Gene, Nuecin (곤충세포주에서 누에신 단백질의 발현 및 성상구명)

  • 윤은영;구태원;황재삼;김상현;강석우;김근영;진병래
    • Journal of Sericultural and Entomological Science
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    • v.44 no.2
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    • pp.64-68
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    • 2002
  • The antibacterial protein gene, nuecin was expressed in Sf9 cells using baculovirus expression vector system (BEVS). The antibactetial activity of mature nuecin against Pectobacterium carotovorum subsp. carotovorum, Ralstonia solanacearum and Pseudomonas tolaasii was significantly high, demonstrating that nuecin had a wider antibacterial spectrum on gram negative and positive bacteria. The result appears to be superior to other antibacterial peptide, attacin. The nuecin was purified by SP-sepharose and Mono Q HR ion-exchange chromatography, and then by Superdex 200 HR 10/30 column. The purified nuecin is quite stable at 80$\^{C}$ and 100$\^{C}$ for several hours of incubation and in a wide pH range (pH 2-12).

Molecular Cloning and Expression of the Novel Attacin-Like Antibacterial Protein Gene Isolated from the Bombyx mori (집누에로부터 새로운 attactin 유산 항세균성 펩타이드 유전자의 분리 및 발현)

  • 윤은영;김상현;강석우;진병래;김근영;김호락;한명세;강석권
    • Korean journal of applied entomology
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    • v.36 no.4
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    • pp.331-340
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    • 1997
  • Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

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Bacillus polyfermenticus CJ9, Isolated from Meju, Showing Antifungal and Antibacterial Activities (메주로부터 분리한 항진균 및 항세균 활성의 Bacillus polyfermenticus CJ9)

  • Jung, Ji-Hye;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.340-349
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    • 2009
  • A CJ9 bacterial strain, which showed antifungal and antibacterial activities, was isolated from meju and identified as Bacillus polyfermenticus based on Gram staining, biochemical properties, as well as its 16S rRNA sequence. B. polyfermenticus CJ9 showed the antimicrobial activity against the various pathogenic molds, yeasts, and bacteria. The antibacterial activity was stable in the pH 5.0~9.0, but the activity was lost at $37^{\circ}C$ for 24 hr. The antifungal activity was stable in the pH range of 3.0~9.0 and reduced at $121^{\circ}C$ for 15 min, but antifungal activity was not completely destroyed. The antibacterial activity was completely inactivated by proteinase K, protease, trypsin, and $\alpha$-chymotrypsin. The antifungal activity was also completely inactivated by protease and $\alpha$-chymotrypsin, and reduced its activity by proteinase which indicated that the antifungal and antibacterial compounds have proteineous nature. The apparent molecular mass of the partially purified antifungal compound, as indicated by using the direct detection method in Tricine-SDS-PAGE, was approximately 1.4 kDa. The molecular mass of the antibacterial compound could not be determined because of its heat-liable characteristic.

An Antibacterial Lectin from Lampteromyces japonicus (화경버섯의 항세균성 렉틴)

  • Yoon, Joo-Ok;Min, Tae-Jin;Yoon, Hee-Sik
    • The Korean Journal of Mycology
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    • v.23 no.1 s.72
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    • pp.46-52
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    • 1995
  • A lectin was isolated from the fruiting bodies of Lampteromyces japonicus by preparative PAGE and named LJAP (Lampteromyces japonicus antibacterial protein). LJAP was a polymeric protein of more than one hundred kDa consisting of 17-kDa subunits. The amino acid analysis revealed a high content of serine, glycine, and acidic amino acids. LJAP has an excellent antibacterial activity for Escherichia coli, JM 109, K 12, HB 101, and JW 380. By the inhibition assay of the antibacterial activity, a glycoprotein, asialofetuin was confirmed as the best inhibitor. This is the first lectin isolated and characterized its antibacterial and agglutination activities from the family Lampteromyces.

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Characterization of Bacteriocin-Like Substances Produced by Bacillus subtilis MJP1 (Bacillus subtilis MJP1이 생산하는 Bacteriocin-Like Substances)

  • Yang, Eun-Ju;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.339-346
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    • 2007
  • The MJP1 bacterial strain, which possesses antifungal activity, was isolated from meju and identified as Bacillus subtilis based on its morphological and biochemical properties, as well as its 16S rRNA sequence. Antimicrobial activity was found against various species of Gram-positive bacteria, yeasts, and molds, including food-spoilage microorganisms. The antifungal activity was found to be stable after heat and proteolytic enzyme treatment, and in the pH range of $6.0{\sim}10.0$. The antibacterial activity was stable in the pH range of $6.0{\sim}10.0$, but about 50% of the activity was lost after 24 hr at $30^{\circ}C$. The antibacterial compound was also inactivated by proteolytic enzyme treatment, indicating its proteinaceous nature. The apparent molecular masses of the partially purified antifungal and antibacterial compounds, as indicated by using the direct detection method in Tricine-SDS-PAGE, were approximately 2.4 kDa and 4.5 kDa, respectively. These studies suggest that B. subtilis MJP1 produces two bacteriocin-like substances with antifungal and antibacterial activities.

Expression of Attacin-like Antibacterial Protein NUECIN in Pleurotus ostreatus

  • Kim, Beom-Gi;Yun, Eun-Young;Yoo, Youngbok
    • Journal of Mushroom
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    • v.9 no.3
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    • pp.91-95
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    • 2011
  • Pleurotus ostreatus, the oyster mushroom, is one of the most important edible mushrooms. It is especially susceptible to bacterial blotch disease, which is caused by Pseudomonas tolaasii. In order to develop bacterial blotch disease-resistant transgenic mushroom, NUECIN cDNA, a gene for an antibacterial peptide cloned from Bombyx mori, was overexpressed in Pleurotus ostreatus. NUECIN cDNA was fused to the ${\beta}$-TUBULIN promoter of oyster mushroom and co-transformed with the pTRura3-2 vector into the uracil auxotrophic mutant strain. Twelve transformants containing the NUECIN gene were identified by genomic PCR and Southern blot analysis. NUECIN gene expression was confirmed by Northern blot analysis. Three transformants showed the transcriptional expression of the gene. However, we could not detect expression of the protein in the transformants. This study showed the possibility of transgenic mushroom development for disease resistance.

Characterization of Antibacterial Compounds from Bacillus polyfermenticus CJ6 and Its Growth Inhibition Effect on Food-Borne Pathogens (Bacillus polyfermenticus CJ6가 생산하는 항세균 물질의 특성 및 병원성 식중독 미생물의 성장 억제 효과)

  • Jung, Ji-Hye;Chang, Hae-Choon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.6
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    • pp.903-911
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    • 2011
  • In this study, Bacillus polyfermenticus CJ6 harboring antibacterial activity was isolated from meju. The antibacterial activity of Bacillus polyfermenticus CJ6 was stable in the pH range of 3.0~9.0, but it disappeared after culture at $70^{\circ}C$ for 24 hr. Antibacterial activity was inactivated by proteinase K, protease, and ${\alpha}$-chymotrypsin, indicating its proteinaceous nature. The growth inhibitory effects of B. polyfermenticus CJ6 culture on food-borne pathogens such as Staphylococcus aureus, Salmonella Typhi, Listeria monocytogenes, and Escherichia coli O157:H7 were examined in this study. Approximately 6~6.2 log CFU/mL of each pathogen was co-cultured with B. polyfermenticus CJ6 in a 50 mL culture volume for 24 hr. Growth of S. aureus and L. monocytogenes was completely inhibited after 3 hr of incubation. Growth of S. Typhi and E. coli O157:H7 was also completely inhibited after 6 hr of incubation. The antibacterial compounds from B. polyfermenticus CJ6 were purified by solid phase extraction (C18 Sep-pak cartridge), recycling preparative HPLC, and analytical HPLC. Ultra-high performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compounds, which were confirmed to be five peptides (757.4153 Da, 750.3444 Da, 1024.5282 Da, 1123.6083 Da, and 1617.8170 Da).

천마를 첨가한 두부의 품질 특성변화

  • 김진성;이준호;하영선
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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    • pp.162.1-162
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    • 2003
  • 천마(Gastrodia Rhizoma)는 뽕나무 버섯과 편리공생하는 난초과(Orcbidan ceae)에 속하는 다년생 초본인 천마(수자해좇, 정풍초, Gastrodia elata Blume)의 뿌리를 지칭하는 것으로서 잎이 없으며, 땅속의 괴경이 한약재로 이용된다. 주로 고혈압, 신경성질환, 당뇨병 등의 성인병 뿐 아니라 스트레스해소, 피로회복 등에 효과가 있는 것으로 알려져 있는 한약재이다. 천마의 주요 효능으로는 항경련 작용 및 항간질 작용에 대한 보고가 있으며, 약리작용을 나타내는 주요성분은 gastrodin, p-hydroxy benzyl alcohol, p-hydroxy benzyl aldehyde, vanillin등의 ployphenol성분으로 알려지고 있다. 한편 두부는 동물성 단백질 식품과는 다르게 열량과 포화지방 함량이 낮고 콜레스테롤이 함유되어 있지 않으며 ,가격이 저렴하여 경제적인 단백질 식품으로 각광받고 있다. 따라서 임상적, 약리적 효능이 있는 물질을 강화하여 기존두부에 비해 영양학적인 기능성을 향상시킬 목적으로 두유에 천마즙을 혼합하여 제조하였고, 기능성 두부제조시 가수량, 가열시간 및 천마즙의 첨가량이 두부수율에 미치는 영향과 텍스쳐, 색도, 탁도, 세균수 등의 품질특성을 일반 두부와 비교·검토하였다.

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