10 research outputs found
Polycyclic aromatic hydrocarbons: characteristics and its degradation by biocatalysis remediation
An excessive released of polycyclic aromatic hydrocarbons (PAHs) to surroundings is
one of the major factors that cause environmental pollution to increase globally. This
issue had gained scientist’s attention to study PAHs biodegradation pathways and their
toxicity towards humans and the environment. They found that the major mechanism
responsible for the ecological recovery of PAH-contaminated sites happened to be from
the microbial degradation process. However, there are a few limitations faced by the
PAHs degrading bacteria where the bacteria die due to extremely polluted areas. This
leads the researchers to utilize genetic engineering to produce enzymes that can
withstand and survive in extreme environments. Recent information and technology
such as path sources, properties and biochemical pathways by means to produce the
simplest and less harmful components in polluted ecosystems are discussed in this
review. In-depth studies in regards to bacteria biocatalysis involving bacterialproduced-enzymes to degrade PAHs help develop new methods to enhance the
bioremediation effectiveness in the future
Tropomyosin, the major tropical oyster Crassostrea belcheri allergen and effect of cooking on its allergenicity
Characterization of immunoglobulin E-binding proteins (IgE) of scomberomorus commerson lacepede
The objective of this study was to determine the Immunoglobulin E-binding proteins (IgE) and major allergens of Scomberomorus commerson Lacepede(Narrow-barred Spanish mackerel). Allergen extracts were obtained from uncooked and cooked fish by homogenization in phosphate-buffered saline followed by continuous extraction at 4oC or on ice. Protein profiles and IgEbinding patterns were then detected by means of sodium dodecyl polyacrylamidegel electrophoresis (SDS-PAGE) and immunoblotting using sera from patients sensitized to the fish. SDS-PAGE of the uncooked fish extracts revealed 26 protein bands in the range of about 11 to >175 kD, while the cooked extracts produced fewer protein bands. Immunoblotting demonstrated 17 IgE-binding bands, ranging in molecular weight from 11 to 151 kD. Two components with molecular weight of about ~50 and 42 kD showed the highest frequency of IgE-binding
(62.2 and 51.4% respectively) and were identified as the major allergens of this fish allergy. Other IgE-binding proteins including a protein at ~12 kD which was equivalent in size to parvalbumin were identified as the minor
allergens
Characterization of Ras k 1 a novel major allergen in Indian mackerel and identification of parvalbumin as the major fish allergen in 33 Asia‐Pacific fish species
Tropomyosin and Actin Identified as Major Allergens of the Carpet Clam (Paphia textile) and the Effect of Cooking on Their Allergenicity
Objectives. To identify the major allergenic proteins of clam (Paphia textile) and to investigate the effect of different cooking methods on the allergenicity of these identified proteins. Methods. Clam protein extracts were separated by denaturing polyacrylamide gel electrophoresis. IgE reactive proteins were then analyzed by immunoblotting with sera from patients with positive skin prick tests (SPT) to the raw clam extract. Mass spectrometry was used to identify the major allergenic proteins of this clam. Results. Raw extract showed 12 protein bands (18–150 kDa). In contrast, fewer protein bands were seen in the boiled extract; those ranging from 40 to 150 kDa were denatured. The protein profiles were similarly altered by frying or roasting. The immunoblots of raw and boiled extracts yielded 10 and 2 IgE-binding proteins, respectively. The fried and roasted extracts showed only a single IgE-binding protein at 37 kDa. Mass spectrometry analysis of the 37 and 42 kDa major allergens indicated that these spots were tropomyosin and actin, respectively. Conclusion. The two major allergens of Paphia textile were identified as the thermostable tropomyosin and a new thermolabile allergen actin
Identification of Major and Minor Allergens of Black Tiger Prawn ( Penaeus monodon ) and King Prawn ( Penaeus latisulcatus )
Background: Prawns and shrimp are a frequent cause of seafood allergy
mediated by IgE antibodies. Penaeus monodon and Penaeus latisulcatus
, commonly known as black tiger prawn and king prawn, respectively, are
among the most frequently consumed prawns in Malaysia. The aim of thi s
study was to identify the IgE-binding proteins of these 2 prawn
species. Methods: Raw and boiled prawn extracts were prepared and then
resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis
(SDS-PAGE). IgE-immunoblotting was then performed using sera from
patients with positive skin prick tests to the raw prawn extracts.
Results: SDS-PAGE analysis of the raw extracts of both prawn species
revealed 23 protein bands; the boiled extracts yielded fewer protein
bands. The bands in the range of 40 to 100 kDa were sensitive to heat
and therefore were not found in the boiled extracts. Immunoblot of raw
extracts of black tiger prawns and king prawns yielded 14 and 11
IgE-binding proteins, respectively, with molecular weights of between
15 and 200 kDa. Proteins at 36, 42, and 49 kDa were detected as the
major allergens in both species of prawns. A protein of 75 kDa was also
identified as a major allergen in black tiger prawns. Other potential
allergens were also observed at various molecular masses. Conclusion:
Proteins of 36, 42, and 49 kDa were identified as the major allergens
of both species of prawns. The 36 and 42 kDa proteins are hypothesised
to be tropomyosin and arginine kinase, respectively. A high molecular
weight protein of 75 kDa was found to be an additional major allergen
in black tiger prawns
Analysis of Pseudomonas sp. polycyclic aromatic hydrocarbons degradation proteins by LC-MS/MS
The uncontrolled release of Polycyclic Aromatic Hydrocarbons (PAHs) had warned of some environmental issues. A bioremediation method by introducing PAHs-degraders bacteria into the contaminated area had issued with a few limitations including survival of strains and against international legislation which we believed that the problems can be reduced with biocatalysis remediation strategies. Biocatalysis of PAHs was detected from biostimulated-PAHs degrading enzymes (PDEs) from Pseudomonas putida strain B4 (PPB4) (Figure 1). Extracellular PDEs were determined by production of 9,10 anthraquinone from PAH biodegradation assay. 2DE analysis was done to compare the expression level of biostimulated and non-stimulated protein. A spot was detected as an upregulated protein while 3 spots were found to be downregulated proteins. These spots undergo LC-MS analysis for protein identification purposes utilizing SEQUEST HT software against the most recent Uniprot database. The results revealed that the upregulated proteins are chaperonin protein (WP_010952474.1), ATP synthase subunit beta (WP_003253197.1) and two dehydrogenase proteins, which are dihydrolipoyl dehydrogenase (WP_010954949.1) and malate dehydrogenase (WP_004575488.1). While downregulated proteins were identified as azurin (WP_003249580.1) and cold-shock protein (WP_003250656.1). In future, these proteins will be further expressed and tested for their ability to work together to achieve the best enzymes consortium for biocatalysis remediation