MfLIP1, a gene encoding an extracellular lipase of the lipid-dependent fungus Malassezia furfur

Malassezia furfur is a dimorphic fungus and a member of the normal cutaneous microflora of humans. However, it is also a facultative pathogen, associated with a wide range of skin diseases. One unusual feature of M. furfur is an absolute dependency on externally provided lipids which the fungus hydrolyses by lipolytic activity to release fatty acids necessary for both growth and pathogenicity. In this study, the cloning and characterization of the first gene encoding a secreted lipase of M. furfur possibly associated with this activity are reported. The gene, MfLIP1, shows high sequence similarity to other known extracellular lipases, but is not a member of a lipase gene family in M. furfur. MfLIP1 consists of 1464 bp, encoding a protein with a molecular mass of 54·3 kDa, a conserved lipase motif and an N-terminal signal peptide of 26 aa. By using a genomic library, two other genes were identified flanking MfLIP1, one of them encoding a putative secreted catalase, the other a putative amine oxidase. The cDNA of MfLIP1 was expressed in Pichia pastoris and the biochemical properties of the recombinant lipase were analysed. MfLip1 is most active at 40 °C and the pH optimum was found to be 5·8. The lipase hydrolysed lipids, such as Tweens, frequently used as the source of fatty acids in M. furfur media, and had minor esterase activity. Furthermore, the lipase is inhibited by different bivalent metal ions. This is the first molecular description of a secreted lipase from M. furfur.

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Characterization of the Gene Encoding the Major Secreted Lysophospholipase A of Legionella pneumophila and Its Role in Detoxification of Lysophosphatidylcholine

Infection and Immunity ◽

10.1128/iai.70.11.6094-6106.2002 ◽

2002 ◽

Vol 70 (11) ◽

pp. 6094-6106 ◽

Cited By ~ 77

Author(s):

Antje Flieger ◽

Birgid Neumeister ◽

Nicholas P. Cianciotto

Keyword(s):

Fatty Acids ◽

Legionella Pneumophila ◽

Lipolytic Activity ◽

Cholesterol Acyltransferase ◽

Phospholipase A ◽

Wild Type ◽

Gene Encoding ◽

Acyltransferase Activity ◽

Lipolytic Enzymes ◽

Cloned Gene

ABSTRACT We previously showed that Legionella pneumophila secretes, via its type II secretion system, phospholipase A activities that are distinguished by their specificity for certain phospholipids. In this study, we identified and characterized plaA, a gene encoding a phospholipase A that cleaves fatty acids from lysophospholipids. The plaA gene encoded a 309-amino-acid protein (PlaA) which had homology to a group of lipolytic enzymes containing the catalytic signature GDSL. In Escherichia coli, the cloned gene conferred trypsin-resistant hydrolysis of lysophosphatidylcholine and lysophosphatidylglycerol. An L. pneumophila plaA mutant was generated by allelic exchange. Although the mutant grew normally in standard buffered yeast extract broth, its culture supernatants lost greater than 80% of their ability to release fatty acids from lysophosphatidylcholine and lysophosphatidylglycerol, implying that PlaA is the major secreted lysophospholipase A of L. pneumophila. The mutant's reduced lipolytic activity was confirmed by growth on egg yolk agar and thin layer chromatography and was complemented by reintroduction of an intact copy of plaA. Overexpression of plaA completely protected L. pneumophila from the toxic effects of lysophosphatidylcholine, suggesting a role for PlaA in bacterial detoxification of lysophospholipids. The plaA mutant grew like the wild type in U937 cell macrophages and Hartmannella vermiformis amoebae, indicating that PlaA is not essential for intracellular infection of L. pneumophila. In the course of characterizing plaA, we discovered that wild-type legionellae secrete a phospholipid cholesterol acyltransferase activity, highlighting the spectrum of lipolytic enzymes produced by L. pneumophila.

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Cloning and Characterization of the Gene Encoding the Major Cell-Associated Phospholipase A of Legionella pneumophila, plaB, Exhibiting Hemolytic Activity

Infection and Immunity ◽

10.1128/iai.72.5.2648-2658.2004 ◽

2004 ◽

Vol 72 (5) ◽

pp. 2648-2658 ◽

Cited By ~ 51

Author(s):

Antje Flieger ◽

Kerstin Rydzewski ◽

Sangeeta Banerji ◽

Markus Broich ◽

Klaus Heuner

Keyword(s):

Legionella Pneumophila ◽

Hemolytic Activity ◽

Genomic Library ◽

Lipolytic Activity ◽

Acanthamoeba Castellanii ◽

Intracellular Survival ◽

Phospholipase A ◽

Wild Type ◽

Gene Encoding ◽

Human Red Blood Cells

ABSTRACT Legionella pneumophila, the causative agent of Legionnaires' disease, is an intracellular pathogen of amoebae, macrophages, and epithelial cells. The pathology of Legionella infections involves alveolar cell destruction, and several proteins of L. pneumophila are known to contribute to this ability. By screening a genomic library of L. pneumophila, we found an additional L. pneumophila gene, plaB, which coded for a hemolytic activity and contained a lipase consensus motif in its deduced protein sequence. Moreover, Escherichia coli harboring the L. pneumophila plaB gene showed increased activity in releasing fatty acids predominantly from diacylphospho- and lysophospholipids, demonstrating that it encodes a phospholipase A. It has been reported that culture supernatants and cell lysates of L. pneumophila possess phospholipase A activity; however, only the major secreted lysophospholipase A PlaA has been investigated on the molecular level. We therefore generated isogenic L. pneumophila plaB mutants and tested those for hemolysis, lipolytic activities, and intracellular survival in amoebae and macrophages. Compared to wild-type L. pneumophila, the plaB mutant showed reduced hemolysis of human red blood cells and almost completely lost its cell-associated lipolytic activity. We conclude that L. pneumophila plaB is the gene encoding the major cell-associated phospholipase A, possibly contributing to bacterial cytotoxicity due to its hemolytic activity. On the other hand, in view of the fact that the plaB mutant multiplied like the wild type both in U937 macrophages and in Acanthamoeba castellanii amoebae, plaB is not essential for intracellular survival of the pathogen.

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Complex processing of lipids from liver of the gonatid squid Berryteuthis magister

Izvestiya TINRO ◽

10.26428/1606-9919-2014-176-34-41 ◽

2014 ◽

Vol 176 (1) ◽

pp. 288-294

Author(s):

Ekaterina V. Ermolenko ◽

Ruslan M. Sultanov ◽

Sergey P. Kasyanov ◽

Yury G. Blinov

Keyword(s):

Fatty Acids ◽

Skin Diseases ◽

Malignant Tumors ◽

Raw Materials ◽

Senile Dementia ◽

North West ◽

The North ◽

Radio Therapy ◽

Wide Range ◽

Main Component

Berryteuthis magister is one of the most common commercial species in the North-West Pacific, but its liver never was utilized before, though it was a rich source of the lipids with simple ether links, 1- О -alkyl-glycerol ethers (АGE), and ω3 polyunsaturated fatty acids (ω3 PUFA). Technology for complex processing of lipids from its liver is developed and presented in details. These forms of lipids are distinguished by a wide range of biologic activity: AGE are effective immunomodulators which enhance hemopoiesis, relieve lesions and lower risk of secondary lesions under radio-therapy and can be used for treatment of some malignant tumors (glioma, prostate cancer, lung carcinoma, etc.); ω3 PUFA could be used for prophylactic of cardiovascular diseases, Alzheimer’s sclerosis, senile dementia, skin diseases (protein synthesis control) and other diseases. The presented technology provides extraction of three preparations with different chemical composition and pharmacological activity. Both saturated and unsaturated AGE could be extracted using the property of different solubility in organic solvents for different components of the mixture of hydrolyzed lipids. The main component of the saturated AGE fraction is chimyl alcohol (90.5 %), but the unsaturated fraction contains 54.7 % of monounsaturated AGE. The ω3 PUFA are concentrated to summary 46 % content of eicosapentaenoic acid and docosahexaenoic acid by the method of fatty acids crystallization with carbamide. Comparing with traditional raw materials for АGE and ω3 PUFA production, as shark liver, the liver of gonatid squid is more suitable because of its high and accessible resources, whereas sharks are presented in bycatch only so cannot be considered as a stable source.

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Fatty Acids and Inflammatory Skin Diseases

Nishi Nihon Hifuka ◽

10.2336/nishinihonhifu.80.5 ◽

2018 ◽

Vol 80 (1) ◽

pp. 5-8

Author(s):

Tetsuya HONDA

Keyword(s):

Fatty Acids ◽

Skin Diseases ◽

Inflammatory Skin Diseases ◽

Inflammatory Skin

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Soames' & Southam's Oral Pathology

10.1093/oso/9780199697786.001.0001 ◽

2018 ◽

Author(s):

Max Robinson ◽

Keith Hunter ◽

Michael Pemberton ◽

Philip Sloan

Keyword(s):

Skin Diseases ◽

Oral Pathology ◽

Pathological Features ◽

Oral Diseases ◽

Oral Medicine ◽

Cross Sectional ◽

Facial Region ◽

Wide Range ◽

Cross Sectional Imaging ◽

Key Aspects

A sound understanding of clinical oral pathology is essential if a dental clinician is to navigate successfully through clinical guidelines, make timely referrals to specialists, and provide good care for patients. This new edition of Soames' & Southam's Oral Pathology provides a clear and friendly guide for students, practitioners, and the whole dental team. Thoroughly updated for today's clinical practice, this textbook covers 'must-know' oral pathology and integrates key aspects of oral medicine. It begins by explaining the principles of clinical assessment, the synthesis of a differential diagnosis, and the selection of further investigations including laboratory tests. Ten chapters bring this theory to life by looking at the clinical and pathological features of a wide range of common oral diseases including oral cancer, salivary gland disorders, and diseases of the jaws. Two new chapters address skin diseases affecting the oro-facial region and neck lumps. A final chapter highlights the importance of clinical oral pathology in the context of systemic human disease. New radiology content includes examples of cross-sectional imaging. Photomicrographs have been replaced with carefully selected images to illustrate key pathological features. Each chapter includes key points boxes and tables to aid learning. Written by experts in both oral pathology and oral medicine, this new edition is a must-have for dentistry students, and those working in the field, providing current and trustworthy information.

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Lipolytic Activity of Liver Tissue of Rats on a Diet Deficient in Essential Fatty Acids

American Journal of Physiology-Legacy Content ◽

10.1152/ajplegacy.1953.174.1.49 ◽

1953 ◽

Vol 174 (1) ◽

pp. 49-50 ◽

Cited By ~ 2

Author(s):

M. Osman Saka ◽

Ümran Sipahioglu

Keyword(s):

Fatty Acids ◽

Liver Tissue ◽

Lipolytic Activity ◽

Essential Fatty Acids

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Optimization Model of Phenolics Encapsulation Conditions for Biofortification in Fatty Acids of Animal Food Products

Foods ◽

10.3390/foods10040881 ◽

2021 ◽

Vol 10 (4) ◽

pp. 881

Author(s):

Roberta Tolve ◽

Fernanda Galgano ◽

Nicola Condelli ◽

Nazarena Cela ◽

Luigi Lucini ◽

...

Keyword(s):

Fatty Acids ◽

Condensed Tannins ◽

Encapsulation Efficiency ◽

Biological Effects ◽

Fatty Acid Content ◽

Food Products ◽

Animal Origin ◽

Animal Nutrition ◽

Loading Capacity ◽

Wide Range

The nutritional quality of animal products is strongly related to their fatty acid content and composition. Nowadays, attention is paid to the possibility of producing healthier foods of animal origin by intervening in animal feed. In this field, the use of condensed tannins as dietary supplements in animal nutrition is becoming popular due to their wide range of biological effects related, among others, to their ability to modulate the rumen biohydrogenation and biofortify, through the improvement of the fatty acids profile, the derivate food products. Unfortunately, tannins are characterized by strong astringency and low bioavailability. These disadvantages could be overcome through the microencapsulation in protective matrices. With this in mind, the optimal conditions for microencapsulation of a polyphenolic extract rich in condensed tannins by spray drying using a blend of maltodextrin (MD) and gum Arabic (GA) as shell material were investigated. For this purpose, after the extract characterization, through spectrophotometer assays and ultra-high-performance liquid chromatography-quadrupole time-of-flight (UHPLC-QTOF) mass spectrometry, a central composite design (CCD) was employed to investigate the combined effects of core:shell and MD:GA ratio on the microencapsulation process. The results obtained were used to develop second-order polynomial regression models on different responses, namely encapsulation yield, encapsulation efficiency, loading capacity, and tannin content. The formulation characterized by a core:shell ratio of 1.5:5 and MD:GA ratio of 4:6 was selected as the optimized one with a loading capacity of 17.67%, encapsulation efficiency of 76.58%, encapsulation yield of 35.69%, and tannin concentration of 14.46 g/100 g. Moreover, in vitro release under varying pH of the optimized formulation was carried out with results that could improve the use of microencapsulated condensed tannins in animal nutrition for the biofortification of derivates.

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Molecular Cloning and Characterization of Bifidobacterium bifidum 1,2-α-l-Fucosidase (AfcA), a Novel Inverting Glycosidase (Glycoside Hydrolase Family 95)

Journal of Bacteriology ◽

10.1128/jb.186.15.4885-4893.2004 ◽

2004 ◽

Vol 186 (15) ◽

pp. 4885-4893 ◽

Cited By ~ 154

Author(s):

Takane Katayama ◽

Akiko Sakuma ◽

Takatoshi Kimura ◽

Yutaka Makimura ◽

Jun Hiratake ◽

...

Keyword(s):

Amino Acid ◽

Genomic Library ◽

Bifidobacterium Bifidum ◽

Glycoside Hydrolases ◽

Glycoside Hydrolase Family ◽

Amino Acid Residues ◽

Gene Encoding ◽

Sugar Chain ◽

Hydrolysis Of

ABSTRACT A genomic library of Bifidobacterium bifidum constructed in Escherichia coli was screened for the ability to hydrolyze the α-(1→2) linkage of 2′-fucosyllactose, and a gene encoding 1,2-α-l-fucosidase (AfcA) was isolated. The afcA gene was found to comprise 1,959 amino acid residues with a predicted molecular mass of 205 kDa and containing a signal peptide and a membrane anchor at the N and C termini, respectively. A domain responsible for fucosidase activity (the Fuc domain; amino acid residues 577 to 1474) was localized by deletion analysis and then purified as a hexahistidine-tagged protein. The recombinant Fuc domain specifically hydrolyzed the terminal α-(1→2)-fucosidic linkages of various oligosaccharides and a sugar chain of a glycoprotein. The stereochemical course of the hydrolysis of 2′-fucosyllactose was determined to be inversion by using 1H nuclear magnetic resonance. The primary structure of the Fuc domain exhibited no similarity to those of any glycoside hydrolases (GHs) but showed high similarity to those of several hypothetical proteins in a database. Thus, it was revealed that the AfcA protein constitutes a novel inverting GH family (GH family 95).

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Source of plasma free fatty acids in dogs receiving fat emulsion and heparin

American Journal of Physiology-Legacy Content ◽

10.1152/ajplegacy.1963.204.4.691 ◽

1963 ◽

Vol 204 (4) ◽

pp. 691-695 ◽

Cited By ~ 81

Author(s):

H. C. Meng ◽

B. Edgren

Keyword(s):

Fatty Acids ◽

Free Fatty Acids ◽

Lipolytic Activity ◽

Blocking Agent ◽

Fat Emulsion ◽

Similar Reduction ◽

Plasma Free Fatty Acids ◽

Fat Infusion ◽

Plasma Ffa ◽

Slight Rise

Unanesthetized dogs were given either 3.0 g fat/kg as a 20% fat emulsion or heparin (2 mg/kg) intravenously or both. Plasma free fatty acids (FFA) and lipolytic activity were determined at intervals. In some experiments hexamethonium (5 mg/kg), a sympathetic ganglionic blocking agent, was administered intravenously either before or after fat or heparin. In fasting dogs fat infusion produced a moderate and heparin caused a slight rise in plasma FFA. Heparin given during lipemia produced a marked elevation of plasma FFA. The plasma lipolytic activity was increased after fat emulsion or heparin. Hexamethonium reduced the fasting plasma FFA about 70% or 0.40–0.6 mEq/liter. A similar reduction of plasma FFA also was observed when hexamethonium was administered during fat infusion or after heparin. Hexamethonium did not affect the increase in plasma lipolytic activity following the administration of fat emulsion or heparin. It seems probable that the increase in plasma FFA observed after intravenous infusion of fat emulsion or heparin is mainly due to the result of intravascular lipolysis.

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