Effect of incubation time and pH on the protein characterization of the aqueous soluble phase of acidified mackerel by-product

SUMMARYCharacterization of the incubation time from infection to onset is important for understanding the natural history of infectious diseases. Attempts to estimate the incubation time distribution for novel A(H1N1v) have been, up to now, based on limited data or peculiar samples. We characterized this distribution for a generic group of symptomatic cases using laboratory-confirmed swine influenza case-information. Estimates of the incubation distribution for the pandemic influenza were derived through parametric time-to-event analyses of data on onset of symptoms and exposure dates, accounting for interval censoring. We estimated a mean of about 1·6–1·7 days with a standard deviation of 2 days for the incubation time distribution in those who became symptomatic after infection with the A(H1N1v) virus strain. Separate analyses for the <15 years and ⩾15 years age groups showed a significant (P<0·02) difference with a longer mean incubation time in the older age group.

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Characterization of Glycoside Hydrolase Family 5 Proteins in Schizosaccharomyces pombe

Eukaryotic Cell ◽

10.1128/ec.00187-10 ◽

2010 ◽

Vol 9 (11) ◽

pp. 1650-1660 ◽

Cited By ~ 11

Author(s):

Encarnación Dueñas-Santero ◽

Ana Belén Martín-Cuadrado ◽

Thierry Fontaine ◽

Jean-Paul Latgé ◽

Francisco del Rey ◽

...

Keyword(s):

Cell Wall ◽

Schizosaccharomyces Pombe ◽

Protein Characterization ◽

Wall Material ◽

Glycoside Hydrolase Family ◽

Content Type ◽

Hydrolysis Of ◽

Controlled Hydrolysis ◽

Hydrolase Family

ABSTRACT In yeast, enzymes with β-glucanase activity are thought to be necessary in morphogenetic events that require controlled hydrolysis of the cell wall. Comparison of the sequence of the Saccharomyces cerevisiae exo-β(1,3)-glucanase Exg1 with the Schizosaccharomyces pombe genome allowed the identification of three genes that were named exg1 + (locus SPBC1105.05), exg2 + (SPAC12B10.11), and exg3 + (SPBC2D10.05). The three proteins have different localizations: Exg1 is secreted to the periplasmic space, Exg2 is a membrane protein, and Exg3 is a cytoplasmic protein. Characterization of the biochemical activity of the proteins indicated that Exg1 and Exg3 are active only against β(1,6)-glucans while no activity was detected for Exg2. Interestingly, Exg1 cleaves the glucans with an endohydrolytic mode of action. exg1 + showed periodic expression during the cell cycle, with a maximum coinciding with the septation process, and its expression was dependent on the transcription factor Sep1. The Exg1 protein localizes to the septum region in a pattern that was different from that of the endo-β(1,3)-glucanase Eng1. Overexpression of Exg2 resulted in an increase in cell wall material at the poles and in the septum, but the putative catalytic activity of the protein was not required for this effect.

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Genomic, Tissue Expression, and Protein Characterization of pCLCA1, a Putative Modulator of Cystic Fibrosis in the Pig

Journal of Histochemistry & Cytochemistry ◽

10.1369/jhc.2009.954594 ◽

2009 ◽

Vol 57 (12) ◽

pp. 1169-1181 ◽

Cited By ~ 15

Author(s):

Stephanie Plog ◽

Lars Mundhenk ◽

Nikolai Klymiuk ◽

Achim D. Gruber

Keyword(s):

Cystic Fibrosis ◽

Tissue Expression ◽

Protein Characterization

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Optimization of Expression Condition, Two Dimensional And Melting Point-Based Characterization of Recombinant Human Interferon Alpha-2a Fusion and Non Fusion Forms

ANNALES BOGORIENSES ◽

10.14203/ann.bogor.2018.v22.n2.57-64 ◽

2018 ◽

Vol 22 (2) ◽

pp. 57

Author(s):

Ratih Asmana Ningrum ◽

Widdya Kusuma Wardhani ◽

Ike Wahyuni ◽

Apon Zaenal Mustopa

Keyword(s):

Interferon Alpha ◽

Protein Characterization ◽

Protein Yield ◽

Size Exclusion ◽

Human Interferon ◽

Two Dimensional ◽

Cancer Treatments ◽

Sds Page ◽

Bca Assay

Recombinant Human Interferon Alpha-2a (rhIFNα-2a) is a therapeutic protein that used in hepatitis and cancer treatments. In our previous research, we developed higher molecular weight of the protein through human serum albumin fusion. The fusion and non fusion form of rhIFNα-2a were produced in Pichia pastoriswith 86 kDa and 19 kDa in size respectively. In previous research, protein yield was not reproducible due to unoptimized expression conditions. This reseach was aimed to optimize expression condition process and to characterize the fusion and non fusion forms of rhIFNα-2a. The parameters to observe in overproduction include nutrient (media and methanol concentration) and non nutrient (temperature andincubation period). Affinity and size exclusion cromatographicwere compared in protein purification. BCA assay was used to determine quantity of protein. Protein characterization was conducted using two-dimensional SDS PAGE and denaturation analyses. The optimal condition of expression was achieved using complex media with 1% of methanol for 3 day incubation period at 25°C. The protein yield was reproducible and higher comparing to previous research. Affinity chromatography resulted in higher purity of the proteins comparing to size exclusions. Characterization using two dimensional gel analysis revealed that isoelectric point of rhIFNα-2a is 6.5 for fusion form and 6.0 for non fusion form. The melting points of fusion protein were 56°C and 62°C whilst that of non fusion was 56°C.

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OPTIMIZATION AND CHARACTERIZATION OF EXO-POLYGALACTURONASE BY Aspergillus niger CULTURED VIA SOLID STATE FERMENTATION

Jurnal Teknologi ◽

10.11113/jt.v81.12222 ◽

2018 ◽

Vol 81 (1) ◽

Author(s):

Halifah Pagarra ◽

Roshanida A. Rahman ◽

Nur Izyan Wan Azelee ◽

Rosli Md Illias

Keyword(s):

Solid State ◽

Aspergillus Niger ◽

Moisture Content ◽

Solid State Fermentation ◽

Incubation Time ◽

Industrial Applications ◽

Screening Process ◽

Industrial Sectors ◽

Polygalacturonase Production

Polygalacturonases represent an important member of pectinases group of enzymes with immense industrial applications. The activity of exo-polygalacturonase produced by Aspergillus niger was studied in solid state fermentation (SSF) using Nephrolepis biserrata leaves as substrate. Central composite design (CCD) was used to optimize four significant variables resulted from the screening process that has been initially analyzed for the production of exo-polygalacturonase which are incubation time, temperature, concentration of pectin and moisture content. The optimum exo-polygalacturonase production obtained was 54.64 U/g at 120 hours of incubation time, temperature at 340C, 5.0 g/L of pectin concentration and 75.26% of moisture content. For partial characterization of exo-polygalacturonase, the optimum temperature and pH were obtained at 50°C and pH 4.0, respectively. SDS-PAGE analysis showed that molecular weight of exo-polygalacturonase were 35 and 71 kDa. This study has revealed a significant production of exo-polygalacturonase by A. niger under SSF using cheap and easily available substrate and thus could found immense potential application in industrial sectors and biotechnology

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A Novel Karyoskeletal Protein: Characterization of Protein NO145, the Major Component of Nucleolar Cortical Skeleton inXenopus Oocytes

Molecular Biology of the Cell ◽

10.1091/mbc.12.12.3904 ◽

2001 ◽

Vol 12 (12) ◽

pp. 3904-3918 ◽

Cited By ~ 8

Author(s):

Sandra Kneissel ◽

Werner W. Franke ◽

Joseph G. Gall ◽

Hans Heid ◽

Sonja Reidenbach ◽

...

Keyword(s):

Divalent Cations ◽

Protein Characterization ◽

Electron Microscopic Level ◽

Expression Library ◽

Electron Microscopic ◽

Egg Formation ◽

Complex Protein ◽

Calculated Mass ◽

Synaptonemal Complex Protein

The nucleolus is a ubiquitous, mostly spheroidal nuclear structure of all protein-synthesizing cells, with a well-defined functional compartmentalization. Although a number of nonribosomal proteins involved in ribosome formation have been identified, the elements responsible for the shape and internal architecture of nucleoli are still largely unknown. Here, we report the molecular characterization of a novel protein, NO145, which is a major and specific component of a nucleolar cortical skeleton resistant to high salt buffers. The amino acid sequence of this polypeptide with a SDS-PAGE mobility corresponding to M r 145,000 has been deduced from a cDNA clone isolated from a Xenopus laevis ovary expression library and defines a polypeptide of 977 amino acids with a calculated mass of 111 kDa, with partial sequence homology to a synaptonemal complex protein, SCP2. Antibodies specific for this protein have allowed its recognition in immunoblots of karyoskeleton-containing fractions of oocytes from differentXenopus species and have revealed its presence in all stages of oogenesis, followed by a specific and rapid degradation during egg formation. Immunolocalization studies at the light and electron microscopic level have shown that protein NO145 is exclusively located in a cage-like cortical structure around the entire nucleolus, consisting of a meshwork of patches and filaments that dissociates upon reduction of divalent cations. We propose that protein NO145 contributes to the assembly of a karyoskeletal structure specific for the nucleolar cortex of the extrachromosomal nucleoli ofXenopus oocytes, and we discuss the possibility that a similar structure is present in other cells and species.

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