• 제목/요약/키워드: Biochemical characterization

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Exoproduction and Biochemical Characterization of a Novel Serine Protease from Ornithinibacillus caprae L9T with Hide-Dehairing Activity

  • Li, Xiaoguang;Zhang, Qian;Gan, Longzhan;Jiang, Guangyang;Tian, Yongqiang;Shi, Bi
    • Journal of Microbiology and Biotechnology
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    • 제32권1호
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    • pp.99-109
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    • 2022
  • This study is the first report on production and characterization of the enzyme from an Ornithinibacillus species. A 4.2-fold increase in the extracellular protease (called L9T) production from Ornithinibacillus caprae L9T was achieved through the one-factor-at-a-time approach and response surface methodological optimization. L9T protease exhibited a unique protein band with a mass of 25.9 kDa upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This novel protease was active over a range of pH (4-13), temperatures (30-80℃) and salt concentrations (0-220 g/l), with the maximal activity observed at pH 7, 70℃ and 20 g/l NaCl. Proteolytic activity was upgraded in the presence of Ag+, Ca2+ and Sr2+, but was totally suppressed by 5 mM phenylmethylsulfonyl fluoride, which suggests that this enzyme belongs to the serine protease family. L9T protease was resistant to certain common organic solvents and surfactants; particularly, 5 mM Tween 20 and Tween 80 improved the activity by 63 and 15%, respectively. More importantly, L9T protease was found to be effective in dehairing of goatskins, cowhides and rabbit-skins without damaging the collagen fibers. These properties confirm the feasibility of L9T protease in industrial applications, especially in leather processing.

코코넛 오일로부터 유래된 당계 비이온 계면활성제 합성 및 계면 특성 연구 (Synthesis of Saccharide Nonionic Biosurfactants from Coconut Oil and Characterization of Their Interfacial Properties)

  • 조선희;이예진;박기호;임종주
    • 공업화학
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    • 제30권4호
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    • pp.435-444
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    • 2019
  • 본 연구에서는 코코넛 오일을 원료로 사용하여 2종의 비이온 당계 생체계면활성제 GP-6 및 GP-7을 합성하였으며, FT-IR, $^1H-NMR$$^{13}C-NMR$ 분광광도계를 이용하여 구조를 규명하였다. 합성한 계면활성제에 대하여 임계 마이셀 농도, 정적 및 동적 표면장력, 계면장력, 유화 작용, 습윤성 및 거품 안정성 등과 같은 계면 물성을 측정한 결과, GP-6 및 GP-7 모두 우수한 계면 특성을 가진 것으로 평가되었다. 또한 Terg-o-tometer를 사용하여 평가한 세척력 측정 결과에 의하면 GP-6 및 GP-7 모두 세제 제품에서 사용되는 기존 계면활성제들과 비교하여 양호한 세척력을 나타내었다. 생분해성, 급성 경구 독성, 급성 피부 자극 및 급성 안자극 검사 결과, GP-6 및 GP-7 모두 저 독성, 저자극성 및 우수한 환경친화성을 가지고 있어서 세제 제품에 적용 가능함을 나타냈으며, 특히 GP-6가 GP-7보다 계면활성이 우수하고 저자극성이고 쉽게 생분해 될 수 있기 때문에 세제 제조에 더 적합할 것으로 판단되었다.

Characterization of Vibrio harveyi, the Causal Agent of Vibriosis in Cultured Marine Fishes in Korea

  • Won, Kyoung-Mi;Kim, Su-Mi;Park, Soo-Il
    • Fisheries and Aquatic Sciences
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    • 제9권3호
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    • pp.123-128
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    • 2006
  • An epizootic causing mortality among cultured marine finfishes occurred in 1999 in the province of Kyoungsang, Korea. The disease was characterized by the presence of enterocele, abdominal swelling, and gastroenteritis. The causative bacteria were isolated from olive flounder (Paralichthys olivaceus), black rockfish (Sebastes schlegeli), turbot (Scophthalmus maximus) and the rearing water. These bacteria showed swarming activity on agar plates and yellowish or greenish colonies on thiosulfate-citrate-bile salts-sucrose (TCBS) agar plates, but no luminescence. The pathogen was identified as Vibrio harveyi based on morphological and biochemical characteristics and the sequence of l6S rDNA. The lethal doses (LD$_{50}$) of olive flounder and black rockfish were estimated to be $1.24\times10^6-1.36\times10^8$ and $3.24\times10^5-5.8\times10^7$ CFU/fish respectively following intraperitoneal injection.

Comparative Genetic Diversity in Natural and Hatchery Populations of Indian Major Carps (C. catla and L. rohita)

  • Rana, R.S.;Bhat, K.V.;Lakhanpal, S.;Lakra, W.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권9호
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    • pp.1197-1203
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    • 2004
  • This study deals with the characterization of three populations (two hatchery and one natural) of Indian major carps Catla catla and Labeo rohita from different locations in India. The genetics of Indian major carps has been completely obscure and this is the first report on comparative allozyme variations in natural and hatchery population. The total 10 biochemical genetic markers used to measure interspecific and intraspecific level of diversity. The allele frequency data indicate different level of genetic variability in three populations. The hatchery population exhibited least polymorphism, low level of heterozygosity and genetic diversity.

Characterization of the Lectin Purified from Canavalia ensiformis Shoots

  • Roh, Kwang-Soo;Park, Na-Young
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권4호
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    • pp.334-340
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    • 2005
  • Lectin is a cell-agglutinating and carbohydrate-binding protein present in many plants. The lectin of Canavalia ensiformis shoot with specific affinity for D-glucose was purified by affinity chromatography using Sephadex G-100, and some of its biochemical characterizations were studied. Lectin was purified 8.87-fold and exhibited final specific activity of 225.74 units/mg protein with a $2.3\%$ yield. SDS-PAGE analysis demonstrated that the purified shoot lectin exists as a tetramer of 102 kD, composed of two subunits with molecular weight of 29 and 22 kD. The purified lectin was observed to agglutinate rabbit blood cell. The optimal temperature for the activity of this lectin was $40^{\circ}C$, and this lectin was relatively stable to heat with the highest activity at $50{\~}60^{\circ}C$. The maximal activity was observed at pH 7.2.

Characterization of 1,4-Benzoquinone Reductase from Bovine Liver

  • Kim, Kyungsoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권4호
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    • pp.216-220
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    • 2002
  • 1,4-Benzoquinone reductase was purified to electrophoretic homogeneity from bovine liver, and the purified enzyme found to have a molecular mass of 29 kDa, as determined by sodium dodecyl sulfate- polyacrylamide gel electrophoresis The enzyme exhibited pH optimum between 8.0 and 8.5. The apparent fm for 1,4-benzoqulnone was 1.643 mM, and the apparent Km for NADH was 1.837 mM. Various divalent cations, such as Hg$\^$2+/, Cu$\^$2+/, and Zn$\^$2+/, exhibited strong inhibitory effects. The enzyme activity was also strongly inhibited by quercetin, dicumarol, and benzoic acid. Incubation of the enzyme with N-bromosuccinimide and pyridoxal 5’-phosphate led to inhibitions of 100% and 99%, respectively. Accordingly, these results suggest that trypto-phan and Iysine residues are Involved at or near the active sites of the enzyme.

Expression, Purification, and Characterization of Prothrombin Kringle 2

  • Rhim, Tai-Youn;Kim, Eun-kyung;Park, Chan-Soo;Kim, Soung-Soo
    • BMB Reports
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    • 제32권2호
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    • pp.147-153
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    • 1999
  • Previously, we reported that the prothrombin kringle 2 (fragment 2), induced by LPS administration into rabbit, inhibited bFGF-stimulated BCE cell growth (Lee et al., 1998). In this study, we cloned and overexpressed the kringle 2 domain of rabbit and human prothrombin as a fusion protein with the pelB leader sequence in E. coli using the T7 promoter. The fusion protein was cleaved during translocation into the peri plasmic space, and cleaved recombinant protein was readily isolated from whole cell lysate by DEAE-Sepharose and Sephacryl S-200 gel filtration chromatography. Both the recombinant rabbit and human prothrombin kringle 2 showed very similar biochemical and functional characteristics to the rabbit prothrombin kringle 2 purified from rabbit serum, in terms of abnormal electrophoretic migration and endothelial cell growth inhibitory activity.

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Isolation and Characterization of Epidermal Mucus from Hirudo nipponia

  • Lee, Ju-Yun;Joe, Cheol-O;Kang, Ke-Won
    • BMB Reports
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    • 제29권3호
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    • pp.248-252
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    • 1996
  • The epidermal organ of the leech contains a complex glycoprotein molecule of mucus. The mucus excreted from annelids plays Significant role in protection against desiccation and parasites. Mucus from the Korean native leech, H. nipponia, was investigated for biochemical characteristics for possible development of biomaterials of cosmetic and pharmaceutical agents. The leech skin mucus was heavily glycosylated mucin-like protein with a high molecular weight comprised 80% carbohydrate and 20% protein. Threonine, serine, and glycine were the major components of the isolated protein and these accounted for 50% of total amino acids. The carbohydrate portion contained glucosamine, galactosamine. galactose, glucose. mannose and sialic acid in oligosaccharide form linked with threonine and serine residues of the glycoprotein.

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Purification and Biochemical Properties of Glutathione S-Transferase from Lactuca sativa

  • Park, Hee-Joong;Cho, Hyun-Young;Kong, Kwang-Hoon
    • BMB Reports
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    • 제38권2호
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    • pp.232-237
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    • 2005
  • A glutathione S-transferase (GST) from Lactuca sativa was purified to electrophoretic homogeneity approximately 403-fold with a 9.6% activity yield by DEAE-Sephacel and glutathione (GSH)-Sepharose column chromatography. The molecular weight of the enzyme was determined to be approximately 23,000 by SDS-polyacrylamide gel electrophoresis and 48,000 by gel chromatography, indicating a homodimeric structure. The activity of the enzyme was significantly inhibited by S-hexylGSH and S-(2,4-dinitrophenyl) glutathione. The enzyme displayed activity towards 1-chloro-2,4-dinitrobenzene, a general GST substrate and high activities towards ethacrynic acid. It also exhibited glutathione peroxidase activity toward cumene hydroperoxide.

길항성 근원 세균이 딸기 시설재배에서 발생하는 잿빛곰팡이병의 생물학적 제어에 미치는 영향 (Effects of Antagonistic Rhizobacteria on the Biological Control of Gray Mold in Greenhouse Grown Strawberry Plants)

  • 조정일;조자용;양승렬
    • 한국유기농업학회지
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    • 제13권2호
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    • pp.161-173
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    • 2005
  • This study was carried out to clarify the effects of antifungal bacterial strains isolated from the greenhouse soil grown strawberry plants on the growth inhibition of plant pathogen, gray mold (Botrytis cinerea) infected in strawberry plants in Damyang and Jangheung districts. Antagonistic bacterial strains were isolated and investigated into the antagonistic activity against gray mold. Screened ten bacterial strains which strongly inhibited Botrytis cinerea were isolated from the greenhouse grown strawberry plants, and the best antifungal microorganism designated as SB 143 was finally selected. Antifungal bacterial strain SB 143 was identified to be the genus Bacillus sp. based on the morphological and biochemical characterization. Bacillus sp. SB 143 showed 59.4% of antifungal activity against Botrytis cinerea. By the bacterialization of culture broth and heated filtrates of culture broth, Bacillus sp. SB 143 showed 93.1% and 32.1% of antagonistic activity against Botrytis cinerea.

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