scholarly journals Overview of the most commonly used methods in allergen characterization

2005 ◽  
Vol 70 (3) ◽  
pp. 347-360 ◽  
Author(s):  
Tanja Cirkovic-Velickovic ◽  
Marija Gavrovic-Jankulovic ◽  
Ratko Jankov
Keyword(s):  
Biochemical Characterization ◽  
Protein Structures ◽  
Protein Molecule ◽  
Underlying Disease ◽  
Type I ◽  
Molecular Weight Determination ◽  
Aminoacid Sequence ◽  
Allergen Molecule

The characterization of an allergen is a troublesome and difficult process, as it requires both the precise biochemical characterization of a (glyco)protein molecule and the establishment of its susceptibility to IgE antibodies, as they are the main link to histamine release in some hypersensitivity states (type I allergies). As the characterization of an allergen includes molecular weight determination of the allergenic molecule, its structure determination, physicochemical properties, IgE binding properties of the allergen molecule, and its allergenicity, an overall review of which biochemical and immunochemical methods are used in achieving this goal are presented in this paper. The information on the molecular level on the structures of allergens indicates that allergens are considerably heterogeneous protein structures, and that there is no particular aminoacid sequence which is responsible for the allergenicity. Therefore, information gained from detailed structural, functional and immunochemical studies of these intriguing molecules, which nowadays modulate a variety of pathophysiological conditions, would greatly improve our understanding of the underlying disease mechanisms, and the way to handle them.

Expression and biochemical characterization of a novel type I pullulanase from Bacillus megaterium

2016 ◽  
Vol 39 (3) ◽  
pp. 397-405 ◽  
Author(s):  
Shaoqing Yang ◽  
Qiaojuan Yan ◽  
Qingdan Bao ◽  
Jingjing Liu ◽  
Zhengqiang Jiang
Keyword(s):  
Bacillus Megaterium ◽  
Biochemical Characterization ◽  
Type I

scholarly journals Genetic and Biochemical Characterization of a Novel Monoterpene ɛ-Lactone Hydrolase from Rhodococcus erythropolis DCL14

2001 ◽  
Vol 67 (2) ◽  
pp. 733-741 ◽  
Author(s):  
Cécile J. B van der Vlugt-Bergmans ◽  
Mariët J. van der Werf
Keyword(s):  
Ring Opening ◽  
Biochemical Characterization ◽  
Rhodococcus Erythropolis ◽  
Open Reading Frames ◽  
Apparent Homogeneity ◽  
Terminal Amino ◽  
Acyl Coenzyme A ◽  
Reading Frames

ABSTRACT A monoterpene ɛ-lactone hydrolase (MLH) from Rhodococcus erythropolis DCL14, catalyzing the ring opening of lactones which are formed during degradation of several monocyclic monoterpenes, including carvone and menthol, was purified to apparent homogeneity. It is a monomeric enzyme of 31 kDa that is active with (4R)-4-isopropenyl-7-methyl-2-oxo-oxepanone and (6R)-6-isopropenyl-3-methyl-2-oxo-oxepanone, lactones derived from (4R)-dihydrocarvone, and 7-isopropyl-4-methyl-2-oxo-oxepanone, the lactone derived from menthone. Both enantiomers of 4-, 5-, 6-, and 7-methyl-2-oxo-oxepanone were converted at equal rates, suggesting that the enzyme is not stereoselective. Maximal enzyme activity was measured at pH 9.5 and 30°C. Determination of the N-terminal amino acid sequence of purified MLH enabled cloning of the corresponding gene by a combination of PCR and colony screening. The gene, designated mlhB(monoterpene lactone hydrolysis), showed up to 43% similarity to members of the GDXG family of lipolytic enzymes. Sequencing of the adjacent regions revealed two other open reading frames, one encoding a protein with similarity to the short-chain dehydrogenase reductase family and the second encoding a protein with similarity to acyl coenzyme A dehydrogenases. Both enzymes are possibly also involved in the monoterpene degradation pathways of this microorganism.

scholarly journals Purification and biochemical characterization of pullulanase type I fromThermus caldophilusGK-24

1996 ◽  
Vol 138 (2-3) ◽  
pp. 147-152 ◽  
Author(s):  
Cheorl-Ho Kim ◽  
Oyekanmi Nashiru ◽  
Jeong Heon Ko
Keyword(s):  
Biochemical Characterization ◽  
Type I

scholarly journals Isolation and Characterization of DNA from Root Tubers

2020 ◽  
pp. 61-67
Author(s):  
O. G. Dawodu ◽  
O. R. Agbeni ◽  
V. Iwu
Keyword(s):  
Gel Electrophoresis ◽  
Manihot Esculenta ◽  
Pcr Amplification ◽  
Colocasia Esculenta ◽  
Marker Genes ◽  
Molecular Weight Determination ◽  
Isolation And Characterization ◽  
Sequencing Method

This study reports a modified SDS extraction technique for isolation of DNA from root tubers namely Dioscorea (yam), Manihot esculenta (cassava) and Colocasia esculenta (cocoyam). DNA extraction in many plants is very difficult because of secondary metabolites that interfere with DNA isolation procedure. Young fresh leaves were collected from the selected tubers and DNA extracted using SDS-extraction protocol, the DNA isolated from the extracted leaves was subjected to gel electrophoresis for determining their purity and concentration and PCR amplification was carried out on the extracted DNA, afterwards gel-electrophoresis was performed for molecular weight determination of the samples. DNA sequencing were performed on both reverse and forward using Sanger sequencing method which gave the nucleotide bases of the samples. DNA isolated by modified SDS protocol method was pure and highest level of purity was obtained from Manihot esculenta, Colocasia esculenta and Dioscorea which are 1.77, 1.72 and 1.20 with concentration of 537.7 ng/µl, 992.4 ng/µl and 149.8 ng/µl respectively. The DNA isolated from root tubers was pure and of good quality, and the isolated DNA was characterized by sequencing. The isolated DNA was successfully sequenced which coded for marker genes of the respective root tubers.

scholarly journals Structural characterization of UDP-galactopyranose mutase from eukaryotic pathogens

2013 ◽  
Author(s):  
◽  
Richa Dhatwalia
Keyword(s):  
Drug Target ◽  
Pathogenic Bacteria ◽  
Leishmania Major ◽  
Biochemical Characterization ◽  
Three Dimensional ◽  
Protozoan Parasites ◽  
Enzymatic Mechanism ◽  
Structural Details

UDP-galactopyranose mutase (UGM) is a unique flavoenzyme that catalyzes the interconversion between UDP-galactopyranose (UDP-Galp) and UDP-galactofuranose (UDP-Galf), without any net transfer of electrons. UGM is a central enzyme involved in the biosynthesis of galactofuranose (Galf). Galf forms a major component of different glycoconjugate structures, lipids and polysaccharides of disease-causing fungi, Aspergillus fumigatus and protozoan parasites such as Trypanosoma cruzi and Leishmania major. Current treatments for diseases caused by these pathogens are limited and use compounds that are either highly toxic or expensive. Thus, new drug development strategies are required for combating these lethal diseases. The unique chemistry of UGMs and its implication in the virulence of pathogenic bacteria, fungi and protozoa and its absence in humans make it a potential drug target. Though bacterial UGMs have been somewhat characterized in detail using structural and biochemical methods, major questions about the catalytic and structural properties of eukaryotic UGMs remain unanswered. Thus, the determination of three-dimensional structures of eukaryotic UGMs might help us in elucidating the enzymatic mechanism of this class of enzymes and potential inhibitor design. The research described in this dissertation address these longstanding questions by providing the first three-dimensional structural details and biochemical characterization of eukaryotic UGMs.

Molecular cloning and biochemical characterization of a heat-stable type I pullulanase from Thermotoga neapolitana

2011 ◽  
Vol 48 (3) ◽  
pp. 260-266 ◽  
Author(s):  
Jinho Kang ◽  
Kyung-Min Park ◽  
Kyoung-Hwa Choi ◽  
Cheon-Seok Park ◽  
Go-Eun Kim ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Biochemical Characterization ◽  
Thermotoga Neapolitana ◽  
Type I ◽  
Stable Type ◽  
Heat Stable
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