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Proteolytic enzymes

L C Perkin, E N Elpidina, B Oppert
Tribolium castaneum is a major agriculture pest damaging stored grains and cereal products. The T. castaneum genome contains 26 cysteine peptidase genes, mostly cathepsins L and B, and seven have a major role in digestion. We targeted the expression of the most highly expressed cathepsin L gene on chromosome 10, TC011001, by RNA interference (RNAi), using double-stranded RNA (dsRNA) constructs of different regions of the gene (3', middle, 5' and entire coding regions). RNA sequencing and quantitation (RNA-seq) was used to evaluate knockdown and specificity amongst the treatments...
October 22, 2016: Insect Molecular Biology
N Rostami, R C Shields, S A Yassin, A R Hawkins, L Bowen, T L Luo, A H Rickard, R Holliday, P M Preshaw, N S Jakubovics
Extracellular DNA (eDNA) has been identified in the matrix of many different monospecies biofilms in vitro, including some of those produced by oral bacteria. In many cases, eDNA stabilizes the structure of monospecies biofilms. Here, the authors aimed to determine whether eDNA is an important component of natural, mixed-species oral biofilms, such as plaque on natural teeth or dental implants. To visualize eDNA in oral biofilms, approaches for fluorescently stained eDNA with either anti-DNA antibodies or an ultrasensitive cell-impermeant dye, YOYO-1, were first developed using Enterococcus faecalis, an organism that has previously been shown to produce extensive eDNA structures within biofilms...
October 21, 2016: Journal of Dental Research
Jan Arends, Nikolas Thomanek, Katja Kuhlmann, Katrin Marcus, Franz Narberhaus
FtsH is the only membrane-bound and essential protease in Escherichia coli. It is responsible for the degradation of regulatory proteins and enzymes like the heat-shock sigma factor RpoH or LpxC, the key enzyme of lipopolysaccharide biosynthesis. To find new FtsH targets, we trapped substrates in E. coli cells from exponential and stationary growth phase using a proteolytically inactive FtsH variant. Subsequent analysis of the isolated FtsH-substrate complexes by label-free nanoLC-MS/MS revealed more than 50 putative FtsH substrates, among them five already known substrates...
October 21, 2016: Proteomics
Samiha Mhamdi, Naourez Ktari, Sawssen Hajji, Moncef Nasri, Alya Sellami Kamoun
The present study was undertaken to characterize the extracellular thermostable serine alkaline proteases from newly actinomycete strain Micromonospora chaiyaphumensis S103 and to describe their evaluation in commercial detergents and shrimp waste deproteinization. This proteolytic crude extract was active and stable in alkaline solution. It was extremely stable in the pH range of 5.0-12.0. The optimum pH and temperature were 8.0 and 70°C, respectively, using casein as a substrate. The thermoactivity and thermostability of proteases were enhanced by the addition of 5mM Ca(2+)...
October 17, 2016: International Journal of Biological Macromolecules
Monalize Salete Mota, Cesar Bauer Gomes, Ismail Teodoro Souza Júnior, Andréa Bittencourt Moura
This study aimed to evaluate the biocontrol potential of bacteria isolated from different plant species and soils. The production of compounds related to phytopathogen biocontrol and/or promotion of plant growth in bacterial isolates was evaluated by measuring the production of antimicrobial compounds (ammonia and antibiosis) and hydrolytic enzymes (amylases, lipases, proteases, and chitinases) and phosphate solubilization. Of the 1219 bacterial isolates, 92% produced one or more of the eight compounds evaluated, but only 1% of the isolates produced all the compounds...
October 4, 2016: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
Aliye Akcalı, Pınar Gümüş, Özgün Özçaka, Banu Öztürk-Ceyhan, Taina Tervahartiala, Heidi Husu, Anna Maria Heikkinen, Timo Sorsa, Nurcan Buduneli
OBJECTIVES: The study evaluated the levels of matrix metalloproteinase-8 (MMP-8), MMP-9, and tissue inhibitor of metalloproteinase-1 (TIMP-1) in biofluids of women with gestational diabetes mellitus (GDM) and systemically healthy counterparts with different status of periodontal health. MATERIALS & METHODS: 71 women with GDM and gingivitis (Gg), 30 women with GDM and healthy periodontium (Gh), 28 systemically and periodontally healthy women (Hh) and 37 systemically healthy women with gingivitis (Hg) were evaluated...
October 20, 2016: Journal of Periodontology
Giuseppe Dionisio, Per Kryger, Tove Steenberg
Beauveria bassiana is an entomopathogenic fungus that grows both in vivo and in vitro. In vivo it can colonize live insect hosts, and tissue digestion occurs by secreted hydrolytic exoenzymes. It can also colonize dead insect tissue provided this is free from competing microorganisms. Depending on whether the host is alive or dead the expression (quality/quantity) of the exoenzymes may vary. We have grown several isolates of B. bassiana in shaking flasks for 120 h at 25 °C in order to evaluate the maximal exoenzyme production using two diet regimes...
October 14, 2016: Insects
Yanhua Cui, Tingting Xu, Xiaojun Qu, Tong Hu, Xu Jiang, Chunyu Zhao
Streptococcus thermophilus is one of the most valuable homo-fermentative lactic acid bacteria, which, for a long time, has been widely used as a starter for the production of fermented dairy products. The key production characteristics of S. thermophilus, for example the production of extracellular polysaccharide, proteolytic enzymes and flavor substances as well as acidifying capacity etc., have an important effect on the quality of dairy products. The acidification capacity of the strains determines the manufacturing time and quality of dairy products...
October 12, 2016: International Journal of Molecular Sciences
Christian Spoerry, Pontus Hessle, Melanie J Lewis, Lois Paton, Jenny M Woof, Ulrich von Pawel-Rammingen
Recently we have discovered an IgG degrading enzyme of the endemic pig pathogen S. suis designated IgdE that is highly specific for porcine IgG. This protease is the founding member of a novel cysteine protease family assigned C113 in the MEROPS peptidase database. Bioinformatical analyses revealed putative members of the IgdE protease family in eight other Streptococcus species. The genes of the putative IgdE family proteases of S. agalactiae, S. porcinus, S. pseudoporcinus and S. equi subsp. zooepidemicus were cloned for production of recombinant protein into expression vectors...
2016: PloS One
Christian Wehner, Klara Janjić, Hermann Agis
Plasmin is a proteolytic enzyme that is crucial in fibrinolysis. In oral tissues, the plasminogen system plays an essential role in physiological and pathological processes, which in addition to fibrinolysis include degradation of extracellular matrix, inflammation, immune response, angiogenesis, tissue remodeling, cell migration, and wound healing. Oral tissues reveal a change in the plasminogen system during pathological processes such as periodontitis, peri-implantitis, or pulpitis, as well as in response to mechanical load...
October 5, 2016: Archives of Oral Biology
Sergei M Danilov, Heinrich Lünsdorf, Henry T Akinbi, Andrew B Nesterovitch, Yuliya Epshtein, Eleftheria Letsiou, Olga V Kryukova, Tobias Piegeler, Elena Z Golukhova, David E Schwartz, Randal O Dull, Richard D Minshall, Olga A Kost, Joe G N Garcia
Angiotensin I-converting enzyme (ACE) hydrolyzes numerous peptides and is a critical participant in blood pressure regulation and vascular remodeling. Elevated tissue ACE levels are associated with increased risk for cardiovascular and respiratory disorders. Blood ACE concentrations are determined by proteolytic cleavage of ACE from the endothelial cell surface, a process that remains incompletely understood. In this study, we identified a novel ACE gene mutation (Arg532Trp substitution in the N domain of somatic ACE) that increases blood ACE activity 7-fold and interrogated the mechanism by which this mutation significantly increases blood ACE levels...
October 13, 2016: Scientific Reports
Lodewijk J A Toonen, Iris Schmidt, Martijn S Luijsterburg, Haico van Attikum, Willeke M C van Roon-Mom
Spinocerebellar ataxia type-3 (SCA3) is a neurodegenerative disorder caused by a polyglutamine repeat expansion in the ataxin-3 protein. Cleavage of mutant ataxin-3 by proteolytic enzymes yields ataxin-3 fragments containing the polyglutamine stretch. These shorter ataxin-3 fragments are thought to be involved in SCA3 pathogenesis due to their increased cellular toxicity and their involvement in formation of the characteristic neuronal aggregates. As a strategy to prevent formation of toxic cleavage fragments, we investigated an antisense oligonucleotide-mediated modification of the ataxin-3 pre-mRNA through exon skipping of exon 8 and 9, resulting in the removal of a central 88 amino acid region of the ataxin-3 protein...
October 12, 2016: Scientific Reports
Yunwi Heo, Young Chul Kwon, Kyoungsoon Shin, Won Duk Yoon, Chang Hoon Han, Seungshic Yum, Euikyung Kim
We have shown that Nemopilema nomurai jellyfish venom (NnV) contains various kinds of proteolytic enzyme activities, including phospholipase (PLA), metalloproteinase (MP) and hyaluronidase activities. In this study, we reported the full-length cDNA and gene sequences of two PLA2 isoforms: acidic PLA2 PA4 and PLA2 PA3A/PA3B/PA5. The full-length cDNA of acidic PLA2 PA4 contains 483 nucleotides (nt), which encode 160 amino acids (and the stop codon), including a signal peptide, six cysteine residues that form disulfide bonds, and metal-binding and catalytic active sites...
October 6, 2016: Toxicon: Official Journal of the International Society on Toxinology
Ailton Cesar Lemes, Gabriel Teixeira Álvares, Mariana Buranelo Egea, Adriano Brandelli, Susana Juliano Kalil
The use of processes for simultaneous production of bioproducts as enzymes and bioactive compounds is an interesting alternative to reduce environmental impacts. Thus, the aim of this study was to produce simultaneously, using the biorefinery concept, both proteases and bioactive compounds with antioxidant activity from Bacillus sp. P45 cultivation by using different by-products. The integrated process developed in this study enabled to obtain enzymes with proteolytic and keratinolytic properties in a process with alternate substrates from agro-industrial by-products (feather meal, residual feather meal and biomass), thus, creating an interesting alternative to managing them...
October 3, 2016: Bioresource Technology
Marta Spodzieja, Katarzyna Kalejta, Aleksandra S Kołodziejczyk, Martyna Maszota-Zieleniak, Sylwia Rodziewicz-Motowidło, Wioletta Żmudzińska, Paulina Czaplewska
Cystatin C originally identified as a cysteine proteases inhibitor has a broad spectrum of biological roles ranging from inhibition of extracellular cysteine protease activities, bone resorption, and modulation of inflammatory responses to stimulation of fibroblasts proliferation. There is an increasing number of evidence to suggest that human cystatin C (hCC) might play a protective role in the pathophysiology of sporadic Alzheimer's disease. In vivo and in vitro results well documented the association of hCC with Aβ and the hCC-induced inhibition of Aβ fibril formation...
October 7, 2016: Journal of Molecular Recognition: JMR
Ying Ji, Shuo Shan, Mingyu He, Chih-Chang Chu
Amino acid-based poly(ester amide)s are a new family of biodegradable polymers that exhibit "pseudo-protein" characteristics and the structural varieties of poly(ester amide)s make them hold great potential in multiple biomedical applications. In this study, a lysine-phenylalanine-based pseudo-protein is developed as the self-assembled nanomicellar carrier for efficient delivery of doxorubicin. The lysine moieties from the pseudo-protein provide available sites for further functionalization, and methylcoumarin is introduced for easy and photocontrollable crosslinking, to effectively improve the micellar stability in serum containing environment and against dilution...
October 6, 2016: Small
Mateusz Adamiak, Malwina Suszynska, Ahmed Abdel-Latif, Ahmed Abdelbaset-Ismail, Janina Ratajczak, Mariusz Z Ratajczak
Migration and bone marrow (BM) homing of hematopoietic stem progenitor cells (HSPCs) is regulated by several signaling pathways, and here we provide evidence for the involvement in this process of hematopoietic-specific phospholipase C-β2 (PLC-β2). This enzyme is involved in release of intracellular calcium and activation of protein kinase C (PKC). Recently we reported that PLC-β2 promotes mobilization of HSPCs from BM into peripheral blood (PB), and this effect is mediated by the involvement of PLC-β2 in the release of proteolytic enzymes from granulocytes and its role in disintegration of membrane lipid rafts...
October 4, 2016: Stem Cell Reviews
S Akram Shirdel, Khosrow Khalifeh, Bijan Ranjbar, Abolfazl Golestani, Khosro Khajeh
We had previously investigated the role of a loop on the activity and conformational stability of chondroitinase ABC Ι (cABC Ι) by constructing some representative mutants in which a network interaction around Asp(689) was manipulated. Here we extended our study by measuring the proteolytic resistance, long term and thermal stability as well as unfolding kinetics of these variants. Long term stability data at 4 and 25°C for 3 weeks indicates that all mutants remain considerably active at 4°C. Thermoinactivation rates for all variants shows that the wild type (WT) enzyme retained 50% of its activity after 2min keeping at 40°C, while L701T, H700N and H700N/L701T as conformationally stabilized variants, have slower inactivation rate...
November 2016: Enzyme and Microbial Technology
Selene Colon, Gautam Bhave
The basement membrane (BM) is a form of extracellular matrix (ECM) which underlies cell layers in nearly all animal tissues. Type IV collagen, a major constituent of BMs, is critical for tissue development and architecture. The enzyme peroxidasin (Pxdn), an ECM associated protein, catalyzes the formation of structurally reinforcing sulfilimine cross-links within the collagen IV network, an event essential to basement membrane integrity. Although the catalytic function of Pxdn is known, the regulation of its activity remains unclear...
October 3, 2016: Journal of Biological Chemistry
Nessma Ahmed El Zawawy, Sameh Samir Ali
INTRODUCTION: Protease enzyme is considered one of the most serious virulence factors produced by extended-spectrum β-lactamase-producing and multidrug-resistant Pseudomonas aeruginosa (ESβLMDRPA) clinical isolates. METHODOLOGY: The antibacterial activity of methanol extract of Ganoderma lucidum fruiting bodies was tested against a protease-producing ESβLMDRPA clinical isolate, showing its mode of action. RESULTS: The extract showed high antibacterial activity...
September 30, 2016: Journal of Infection in Developing Countries
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