A Systematic Analysis and a New Online Resource for Antimicrobial-Enzyme
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Enantioselektive Biotransformationen Zur Synthese Von Molekülen Mit
Untersuchung zur Produktion und gerichteten Immobilisierung einer Alginat Lyase aus Sphingomonas sp. A1 Dissertation zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) vorgelegt der Naturwissenschaftlichen Fakultät I Biowissenschaften der Martin-Luther-Universität Halle-Wittenberg von Herrn Dipl.-Biol. Christian Beyerodt geboren am 15.12.1982 in Halle an der Saale Halle (Saale) 2015 Gutachter/in: 1. Prof. Dr. Markus Pietzsch 2. Prof. Dr. Reinhard Neubert 3. Prof. Dr. Christoph Syldatk Tag der öffentlichen Verteidigung: 30.03.2016 2 Selbstständigkeitserklärung Hiermit erkläre ich, Christian Beyerodt, dass ich die vorliegende Arbeit - mit Ausnahme der aufgeführten Personen, Unterlagen bzw. Literaturstellen - selbstständig und ohne fremde Hilfe angefertigt habe, _________________ Halle, Datum 3 Danksagung Die vorliegende Dissertation entstand während meiner Tätigkeit als wissenschaftlicher Mitar- beiter in der Arbeitsgruppe „Aufarbeitung biotechnischer Produkte“ des Institutes für Phar- mazie an der Martin-Luther-Universität Halle-Wittenberg. Viele Menschen haben mich wäh- rend dieser Zeit unterstützt und dazu beigetragen, dass ich diese Zeit der Promotion als ei- nen positiven Lebensabschnitt in Erinnerung behalten werde. Diesen Personen möchte ich im Folgenden danken. Zuerst möchte ich mich bei Prof. Dr. Markus Pietzsch für das entgegengebrachte Vertrauen, mir das Thema zu überlassen, bedanken. Auch danke ich für die vielen konstruktiven Dis- kussionen, Kritiken und Hilfestellungen, welche diese Arbeit erst ermöglichten. Weiterhin danke ich der Firma Lanxess, im speziellen Frau Dr. Hermsdorf und Herrn Dr. Schellenberg. Das interdisziplinäre Projekt wurde die Grundlage für meine Forschungen. Ich danke natürlich auch der gesamten Arbeitsgruppe „AG-Pietzsch“ für die schöne Zeit und die zahlreichen Aktivitäten neben der Arbeit. Die Sommerfeste waren immer ein riesen Spaß. -
Analysis of Carbohydrates and Glycoconjugates by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry: an Update for 2011–2012
ANALYSIS OF CARBOHYDRATES AND GLYCOCONJUGATES BY MATRIX-ASSISTED LASER DESORPTION/IONIZATION MASS SPECTROMETRY: AN UPDATE FOR 2011–2012 David J. Harvey* Department of Biochemistry, Oxford Glycobiology Institute, University of Oxford, Oxford OX1 3QU, UK Received 19 June 2014; accepted 15 January 2015 Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/mas.21471 This review is the seventh update of the original article published text. As the review is designed to complement the earlier work, in 1999 on the application of MALDI mass spectrometry to the structural formulae, etc. that were presented earlier are not analysis of carbohydrates and glycoconjugates and brings repeated. However, a citation to the structure in the earlier work coverage of the literature to the end of 2012. General aspects is indicated by its number with a prefix (i.e., 1/x refers to such as theory of the MALDI process, matrices, derivatization, structure x in the first review and 2/x to structure x the second). MALDI imaging, and fragmentation are covered in the first part Books, general reviews, and review-type articles directly of the review and applications to various structural types concerned with, or including, MALDI analysis of carbohydrates constitute the remainder. The main groups of compound are and glycoconjugates to have been published during the review oligo- and poly-saccharides, glycoproteins, glycolipids, glyco- period are listed in Table 1. Reviews relating to more specific sides, and biopharmaceuticals. Much of this material is presented aspects of MALDI analysis are listed in the appropriate sections. in tabular form. Also discussed are medical and industrial applications of the technique, studies of enzyme reactions, and II. -
Role of Extracellular Proteases in Biofilm Disruption of Gram Positive
e Engine ym er z in n g E Mukherji, et al., Enz Eng 2015, 4:1 Enzyme Engineering DOI: 10.4172/2329-6674.1000126 ISSN: 2329-6674 Review Article Open Access Role of Extracellular Proteases in Biofilm Disruption of Gram Positive Bacteria with Special Emphasis on Staphylococcus aureus Biofilms Mukherji R, Patil A and Prabhune A* Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune, India *Corresponding author: Asmita Prabhune, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune 411008, India, Tel: 91-020-25902239; Fax: 91-020-25902648; E-mail: [email protected] Rec date: December 28, 2014, Acc date: January 12, 2015, Pub date: January 15, 2015 Copyright: © 2015 Mukherji R, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Bacterial biofilms are multicellular structures akin to citadels which have individual bacterial cells embedded within a matrix of a self-synthesized polymeric or proteinaceous material. Since biofilms can establish themselves on both biotic and abiotic surfaces and that bacteria residing in these complex molecular structures are much more resistant to antimicrobial agents than their planktonic equivalents, makes these entities a medical and economic nuisance. Of late, several strategies have been investigated that intend to provide a sustainable solution to treat this problem. More recently role of extracellular proteases in disruption of already established bacterial biofilms and in prevention of biofilm formation itself has been demonstrated. The present review aims to collectively highlight the role of bacterial extracellular proteases in biofilm disruption of Gram positive bacteria. -
Glycoside Hydrolase Dish from Desulfovibrio Vulgaris Degrades The
Glycoside Hydrolase DisH from Desulfovibrio vulgaris Degrades the N-Acetylgalactosamine Component of Diverse Biofilms Lei Zhu1, Venkata G. Poosarla1, Sooyeon Song1, Thammajun L. Wood1, Daniel S. Miller3, Bei Yin3, and Thomas K. Wood1, 2* 1Department of Chemical Engineering and 2Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA, 16802 3Dow Chemical Company, Collegeville, PA, 19426 *To whom correspondence should be addressed: [email protected] Running title: D. vulgaris DisH disperses its biofilm This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process which may lead to differences between this version and the Version of Record. Please cite this article as an ‘Accepted Article’, doi: 10.1111/1462-2920.14064 This article is protected by copyright. All rights reserved. SIGNIFICANCE The global costs of corrosion are more than $2.5 trillion every year (3.4% of the global gross domestic product), and a large part of corrosion (30%) is microbiologically influenced corrosion (MIC), which affects oil production, drinking water systems, and pipelines. MIC is commonly caused by sulfate- reducing bacteria (SRB) biofilms, and Desulfovibrio vulgaris is the model organism. The biofilm matrix of D. vulgaris consists of proteins and polysaccharides of mannose, fucose, and N-acetylgalactosamine (GalNAc). However, little is known about how to control its biofilm formation. Since bacteria must degrade their biofilm to disperse, in this work, we identified and then studied predicted secreted glycoside hydrolases from D. vulgaris. We show here that DisH (DVU2239, dispersal hexosaminidase), a previously uncharacterized protein, is an N-acetyl-β-D-hexosaminidase that disperses the D. -
Therapeutic Effects of a Combined Antibiotic-Enzyme Treatment on Subclinical Mastitis in Lactating Dairy Cows
Veterinarni Medicina, 61, 2016 (5): 237–242 Original Paper doi: 10.17221/8876-VETMED Therapeutic effects of a combined antibiotic-enzyme treatment on subclinical mastitis in lactating dairy cows B. Khoramian1, M. Emaneini2, M. Bolourchi3, A. Niasari-Naslaji3, A. Gorganzadeh1, S. Abani1, P. Hovareshti3 1Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran 2School of Medicine, Tehran University of Medical Sciences. Tehran, Iran 3Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran ABSTRACT: The objective of this study was to evaluate a combined antibiotic-enzyme therapy for Staphylococcus aureus mastitis and biofilm formation. A total of 141 cases of S. aureus chronic mastitis from three farms were divided ® into two groups: the control group (n = 54) were treated with Nafpenzal ointment; the enzyme + antibiotic group ® ® (n = 87) were treated with Nafpenzal plus an enzymatic ointment (MastiVeyxym ). Quantitative determination of biofilm formation was determined using a colorimetric microplate assay and the detection of ica genes by PCR. Enzyme + antibiotic therapy did not significantly improve cure rates compared to control (48.3 vs 38.9%). The cure rate for infections caused by biofilm-positive strains was 42.6 and 46.6% for control and enzyme + antibiotic groups, respectively (P > 0.05). Comparison of cure rates between farms showed a relationship with somatic cell count (SCC), parity and oxacillin resistance. 79.4% of the isolates produced biofilm and antibacterial resistance rates for oxacillin, penicillin and streptomycin were 25.5, 71.6 and 95.7%, respectively. These results indicate that while the increase in mastitis cure rate using enzymatic therapy was not significant, this treatment could be useful in some situations. -
The Efficacy of Lyticase and Β-Glucosidase Enzymes on Biofilm
Banar et al. BMC Microbiology (2019) 19:291 https://doi.org/10.1186/s12866-019-1662-9 RESEARCH ARTICLE Open Access The efficacy of lyticase and β-glucosidase enzymes on biofilm degradation of Pseudomonas aeruginosa strains with different gene profiles Maryam Banar1, Mohammad Emaneini1, Reza Beigverdi1, Rima Fanaei Pirlar1, Narges Node Farahani1, Willem B. van Leeuwen2 and Fereshteh Jabalameli1* Abstract Background: Pseudomonas aeruginosa is a nosocomial pathogen that causes severe infections in immunocompromised patients. Biofilm plays a significant role in the resistance of this bacterium and complicates the treatment of its infections. In this study, the effect of lyticase and β-glucosidase enzymes on the degradation of biofilms of P. aeruginosa strains isolated from cystic fibrosis and burn wound infections were assessed. Moreover, the decrease of ceftazidime minimum biofilm eliminating concentrations (MBEC) after enzymatic treatment was evaluated. Results: This study demonstrated the effectiveness of both enzymes in degrading the biofilms of P. aeruginosa.In contrast to the lyticase enzyme, β-glucosidase reduced the ceftazidime MBECs significantly (P < 0.05). Both enzymes had no cytotoxic effect on the A-549 human lung carcinoma epithelial cell lines and A-431 human epidermoid carcinoma cell lines. Conclusion: Considering the characteristics of the β-glucosidase enzyme, which includes the notable degradation of P. aeruginosa biofilms and a significant decrease in the ceftazidime MBECs and non-toxicity for eukaryotic cells, this enzyme can be a promising therapeutic candidate for degradation of biofilms in burn wound patients, but further studies are needed. Keywords: Pseudomonas aeruginosa, β-Glucosidase, Lyticase, Biofilm Background patients and associated with chronic infections and antibiotic Pseudomonas aeruginosa is a nosocomial pathogen that resistance [4, 8]. -
Biofilms As Promoters of Bacterial Antibiotic Resistance and Tolerance
antibiotics Review Biofilms as Promoters of Bacterial Antibiotic Resistance and Tolerance Cristina Uruén 1,† , Gema Chopo-Escuin 1,†, Jan Tommassen 2 , Raúl C. Mainar-Jaime 1 and Jesús Arenas 1,* 1 Unit of Microbiology and Immunology, Faculty of Veterinary, University of Zaragoza, Miguel Servet, 177, 50017 Zaragoza, Spain; [email protected] (C.U.); [email protected] (G.C.-E.); [email protected] (R.C.M.-J.) 2 Department of Molecular Microbiology, Institute of Biomembranes, Utrecht University, 3584 CH Utrecht, The Netherlands; [email protected] * Correspondence: [email protected] † These authors contributed equally to this work. Abstract: Multidrug resistant bacteria are a global threat for human and animal health. However, they are only part of the problem of antibiotic failure. Another bacterial strategy that contributes to their capacity to withstand antimicrobials is the formation of biofilms. Biofilms are associations of microorganisms embedded a self-produced extracellular matrix. They create particular environments that confer bacterial tolerance and resistance to antibiotics by different mechanisms that depend upon factors such as biofilm composition, architecture, the stage of biofilm development, and growth condi- tions. The biofilm structure hinders the penetration of antibiotics and may prevent the accumulation of bactericidal concentrations throughout the entire biofilm. In addition, gradients of dispersion of nutrients and oxygen within the biofilm generate different metabolic states of individual cells and favor the development of antibiotic tolerance and bacterial persistence. Furthermore, antimicrobial resistance may develop within biofilms through a variety of mechanisms. The expression of efflux pumps may be induced in various parts of the biofilm and the mutation frequency is induced, while the presence of extracellular DNA and the close contact between cells favor horizontal gene transfer. -
The Role of Serratiopeptidase in the Resolution of Inflammation
ARTICLE IN PRESS asian journal of pharmaceutical sciences ■■ (2017) ■■– ■■ Available online at www.sciencedirect.com ScienceDirect journal homepage: www.elsevier.com/locate/ajps Review The role of serratiopeptidase in the resolution of inflammation Manju Tiwari * Department of Biochemistry and Genetics, Barkatullah University, Bhopal, Madhya Pradesh, India ARTICLE INFO ABSTRACT Article history: Inflammation remains a key event during most of the diseases and physiological imbal- Received 26 October 2016 ance. Acute inflammation is an essential physiological event by immune system for a protective Received in revised form 9 measure to remove cause of inflammation and failure of resolution lead to chronic inflam- December 2016 mation. Over a period of time, a number of drugs mostly chemical have been deployed to Accepted 16 January 2017 combat acute and chronic inflammation. Recently, enzyme based anti-inflammatory drugs Available online became popular over conventional chemical based drugs. Serratiopeptidase, a proteolytic enzyme from trypsin family, possesses tremendous scope in combating inflammation. Serine Keywords: protease possesses a higher affinity for cyclooxygenase (COX-I and COX-II), a key enzyme Inflammation associated with production of different inflammatory mediators including interleukins (IL), Cyclooxygenase prostaglandins (PGs) and thromboxane (TXs) etc. Currently, arthritis, sinusitis, bronchitis, NSAIDs fibrocystic breast disease, and carpal tunnel syndrome, etc. are the leading inflammatory Serratiopeptidase disorders that affected the entire the globe. In order to conquer inflammation, both acute Steroids and chronic world, physician mostly relies on conventional drugs. The most common drugs Enzyme therapeutics to combat acute inflammation are Nonsteroidal anti-inflammatory drugs (NSAIDs) alone and or in combination with other drugs. However, during chronic inflammation, NSAIDs are often used with steroidal drugs such as autoimmune disorders. -
ROLE of in VIVO-INDUCED GENES ILVI and HFQ in the SURVIVAL and VIRULENCE of ACTINOBACILLUS PLEUROPNEUMONIAE By
ROLE OF IN VIVO-INDUCED GENES ILVI AND HFQ IN THE SURVIVAL AND VIRULENCE OF ACTINOBACILLUS PLEUROPNEUMONIAE By Sargurunathan Subashchandrabose A DISSERTATION Submitted to Michigan State University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Comparative Medicine and Integrative Biology 2011 ABSTRACT ROLE OF IN VIVO-INDUCED GENES ILVI AND HFQ IN THE SURVIVAL AND VIRULENCE OF ACTINOBACILLUS PLEUROPNEUMONIAE By Sargurunathan Subashchandrabose Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, a contagious and often fatal disease of pigs. Infection with this bacterium leads to the development of a fulminant pleuropneumonia resulting in severe damage to the lungs. A. pleuropneumoniae genes that are up-regulated during the infection of pig lungs were previously identified in our laboratory and designated as in vivo-induced genes. The role of two such in vivo-induced genes, ilvI and hfq, in the pathobiology of A. pleuropneumoniae is the subject of this dissertation. The gene ilvI encodes an enzyme involved in the biosynthesis of branched-chain amino acids (BCAAs). The leucine-responsive regulatory protein (Lrp) is a transcriptional regulator of the ilvIH operon. BCAA biosynthetic genes are associated with virulence in pathogens infecting the respiratory tract and blood stream. Also, twenty five percent of the A. pleuropneumoniae in vivo-induced genes were up-regulated under BCAA limitation, suggesting that these BCAAs may be found at limiting concentrations in certain sites of the mammalian body such as the respiratory tract. The concentration of amino acids in the porcine pulmonary epithelial lining fluid was determined and BCAAs were found at limiting concentration in the respiratory tract. -
Targeting Microbial Biofilms Using Ficin, a Nonspecific Plant Protease Diana R
www.nature.com/scientificreports OPEN Targeting microbial biofilms using Ficin, a nonspecific plant protease Diana R. Baidamshina1,*, Elena Y. Trizna1,*, Marina G. Holyavka2, Mikhail I. Bogachev3, Valeriy G. Artyukhov2, Farida S. Akhatova1, Elvira V. Rozhina1, Rawil F. Fakhrullin1 & 1 Received: 26 September 2016 Airat R. Kayumov Accepted: 08 March 2017 Biofilms, the communities of surface-attached bacteria embedded into extracellular matrix, are Published: 07 April 2017 ubiquitous microbial consortia securing the effective resistance of constituent cells to environmental impacts and host immune responses. Biofilm-embedded bacteria are generally inaccessible for antimicrobials, therefore the disruption of biofilm matrix is the potent approach to eradicate microbial biofilms. We demonstrate here the destruction ofStaphylococcus aureus and Staphylococcus epidermidis biofilms with Ficin, a nonspecific plant protease. The biofilm thickness decreased two-fold after 24hours treatment with Ficin at 10 μg/ml and six-fold at 1000 μg/ml concentration. We confirmed the successful destruction of biofilm structures and the significant decrease of non-specific bacterial adhesion to the surfaces after Ficin treatment using confocal laser scanning and atomic force microscopy. Importantly, Ficin treatment enhanced the effects of antibiotics on biofilms-embedded cells via disruption of biofilm matrices. Pre-treatment with Ficin (1000 μg/ml) considerably reduced the concentrations of ciprofloxacin and bezalkonium chloride required to suppress the viable Staphylococci by 3 orders of magnitude. We also demonstrated that Ficin is not cytotoxic towards human breast adenocarcinoma cells (MCF7) and dog adipose derived stem cells. Overall, Ficin is a potent tool for staphylococcal biofilm treatment and fabrication of novel antimicrobial therapeutics for medical and veterinary applications. -
Enzyme Dynamics of Biofilm-Breaking Dispersin B
Enzyme Dynamics of Biofilm-Breaking Dispersin B Kim, Edward (School: Kalaheo High School) Biofilm-based infections pose two major problems: extreme resistance to antibiotics and many other conventional antimicrobial agents and extreme capacity for evading the host defenses. Dispersin B is a glycoside hydrolase that hydrolyzes the linear polymers in poly-β-1,6-N-acetylglucosamine, considered the main component of the biofilm. Thus, understanding the enzyme dynamics of biofilm-breaking Dispersin B and the nature of the substrate transition state would be important to develop effective antibiotic strategies. To obtain thermodynamic activation parameters, assays of Dispersin B and a biofilm from Staphylococcus aureus were conducted. Dispersin B successfully disturbed biofilm formation due to its cleaving abilities. Additionally, various concentrations of glucose were added to bacteria media, and addition of glucose was found to improve the biofilm formation. Using the optimal concentration of 8 mg/L glucose, independent temperature-controlled assays were performed with varying degrees of substrate concentration to determine the rate constant, which was input in the Arrhenius and Eyring equations. From the resulting thermodynamic values, the Gibbs free energy of activation was determined to be 62.9 kJ/mol at 37 degrees Celsius. Molecular dynamics simulations were performed to predict the transition state structure. After chemically drawn and optimized using the program Avogadro, the substrate analog (4-Nitrophenyl N-acetyl-β-D-glucosaminide) and the potential transition state structure were docked in the active site of Dispersin B. After 1ns molecular dynamics simulations, the potential transition state was observed. This computational method could yield better models of transition-state inhibitors for drug design. -
Book of Abstracts Of
BOOK OF ABSTRACTS OF BOOK OF ABSTRACTS OF CEB ANNUAL MEETING 2017 6 JULY 2017, BRAGA, PORTUGAL Edited by: Eugénio Campos Ferreira Publisher: Universidade do Minho, Centro de Engenharia Biológica Campus de Gualtar, 4710-057 Braga, Portugal ISBN: 978-989-97478-8-3 DOI: 10.21814/CEBsam2017 © Universidade do Minho This publication contains research works sponsored by Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Foreword The Centre of Biological Engineering (CEB, from the Portuguese title ‘Centro de Engenharia Biológica’) is a research unit of the School of Engineering of the University of Minho, which is recognized as a strategic infrastructure for the development of the Portuguese Biotechnology and Bioengineering fields. The mission of CEB is to generate, disseminate, and apply knowledge with relevance for society in the economic, social and cultural dimensions, and to contribute to the expansion of the scientific and technological fields under its scope of activity. The research carried out at CEB covers the molecular, cellular and process scales, combining knowledge from the exact, natural, health, environmental and engineering sciences, in order to develop new products and processes as well as a wide range of bioengineering and biotechnological applications in the agro-food, environmental, energy, industrial fine chemistry, biomedical and health fields. CEB combines R&D activities with advanced training, technology transfer, consulting and services, with the aim of fostering the industrial and agro-food, health, and environmental sectors.