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  • 1
    Language: English
    In: Biotechnology progress, 2016-03, Vol.32 (2), p.327-336
    Description: This study demonstrates the production of an active enzyme cocktail produced by growing Trichoderma harzianum on sugarcane bagasse. The component enzymes were identified by LCMS‐MS. Glycosyl hydrolases were the most abundant class of proteins, representing 67% of total secreted protein. Other carbohydrate active enzymes involved in cell wall deconstruction included lytic polysaccharide mono‐oxygenases (AA9), carbohydrate‐binding modules, carbohydrate esterases and swollenin, all present at levels of 1%. In total, proteases and lipases represented 5 and 1% of the total secretome, respectively, with the rest of the secretome being made up of proteins of unknown or putative function. This enzyme cocktail was efficient in catalysing the hydrolysis of sugarcane bagasse cellulolignin to fermentable sugars for potential use in ethanol production. Apart from mapping the secretome of T. harzianum, which is a very important tool to understand the catalytic performance of enzyme cocktails, the gene coding for T. harzianum swollenin was expressed in Aspergillus niger. This novel aspect in this work, allowed increasing the swollenin concentration by 95 fold. This is the first report about the heterologous expression of swollenin from T. harzianum, and the findings are of interest in enriching enzyme cocktail with this important accessory protein which takes part in the cellulose amorphogenesis. Despite lacking detectable glycoside activity, the addition of swollenin of T. harzianum increased by two‐fold the hydrolysis efficiency of a commercial cellulase cocktail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:327–336, 2016
    Subject(s): Analysis ; Aspergillus niger ; Biocatalysis ; Cellulases - analysis ; Cellulases - metabolism ; Cellulose ; Cellulose - biosynthesis ; Cellulose - chemistry ; Cellulose - metabolism ; Chemical properties ; enzymatic hydrolysis ; Hydrolysis ; Lipase ; Proteases ; Proteome - chemistry ; Proteome - metabolism ; Saccharum - chemistry ; Saccharum - metabolism ; secretome ; Sugarcane ; sugarcane bagasse ; swollenin ; Trichoderma - chemistry ; Trichoderma - metabolism ; Trichoderma harzianum
    ISSN: 8756-7938
    E-ISSN: 1520-6033
    Source: Wiley Online Library All Backfiles
    Source: Get It Now
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  • 2
    Language: English
    In: Science (American Association for the Advancement of Science), 2012-06-29, Vol.336 (6089), p.1704-1708
    Description: Noscapine is an antitumor alkaloid from opium poppy that binds tubulin, arrests metaphase, and induces apoptosis in dividing human cells. Elucidation of the biosynthetic pathway will enable improvement in the commercial production of noscapine and related bioactive molecules. Transcriptomic analysis revealed the exclusive expression of 10 genes encoding five distinct enzyme classes in a high noscapine-producing poppy variety, HN1. Analysis of an F₂ mapping population indicated that these genes are tightly linked in HN1, and bacterial artificial chromosome sequencing confirmed that they exist as a complex gene cluster for plant alkaloids. Virus-induced gene silencing resulted in accumulation of pathway intermediates, allowing gene function to be linked to noscapine synthesis and a novel biosynthetic pathway to be proposed.
    Subject(s): Alkaloids ; Anticancer properties ; Antineoplastic Agents, Phytogenic - biosynthesis ; Biological and medical sciences ; Biosynthesis ; Capsules ; Classical genetics, quantitative genetics, hybrids ; Clusters ; Cough ; Enzymes ; Flowers & plants ; Fundamental and applied biological sciences. Psychology ; Gene silencing ; Genes ; Genes, Plant ; Genetic aspects ; Genetics of eukaryotes. Biological and molecular evolution ; Genomes ; Latex ; Libraries ; Low level ; Molecular Sequence Data ; Morphinans ; Multigene Family ; Narcotics ; Noscapine - metabolism ; Open reading frames ; Papaver - enzymology ; Papaver - genetics ; Papaver - metabolism ; Papaver somniferum ; Pharmaceutical sciences ; Physiological aspects ; Plant biology ; Poppies ; Pteridophyta, spermatophyta ; REPORTS ; Research ; Synthesis ; Vegetals
    ISSN: 0036-8075
    E-ISSN: 1095-9203
    Source: JSTOR Life Sciences
    Source: Academic Search Ultimate
    Source: Alma/SFX Local Collection
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  • 3
    Language: English
    In: Nature communications, 2018-12-03, Vol.9 (1), p.5125-14
    Description: Woody (lignocellulosic) plant biomass is an abundant renewable feedstock, rich in polysaccharides that are bound into an insoluble fiber composite with lignin. Marine crustacean woodborers of the genus Limnoria are among the few animals that can survive on a diet of this recalcitrant material without relying on gut resident microbiota. Analysis of fecal pellets revealed that Limnoria targets hexose-containing polysaccharides (mainly cellulose, and also glucomannans), corresponding with the abundance of cellulases in their digestive system, but xylans and lignin are largely unconsumed. We show that the limnoriid respiratory protein, hemocyanin, is abundant in the hindgut where wood is digested, that incubation of wood with hemocyanin markedly enhances its digestibility by cellulases, and that it modifies lignin. We propose that this activity of hemocyanins is instrumental to the ability of Limnoria to feed on wood in the absence of gut symbionts. These findings may hold potential for innovations in lignocellulose biorefining.
    Subject(s): Animals ; Cellulase ; Cellulose ; Cellulose - metabolism ; Crustaceans ; Diet ; Digestibility ; Digestion - physiology ; Digestive system ; Digestive tract ; Feces - chemistry ; Fiber composites ; Gastrointestinal Tract - metabolism ; Gastrointestinal Tract - physiology ; Gastrointestinal Tract - ultrastructure ; Hemocyanins ; Hemocyanins - metabolism ; Hexose ; Hindgut ; Incubation ; Innovations ; Isopoda - metabolism ; Isopoda - physiology ; Lignin ; Lignin - metabolism ; Lignocellulose ; Limnoria ; Marine crustaceans ; Marine plants ; Microbiota ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Plant biomass ; Polysaccharides ; Proteins ; Saccharides ; Shellfish ; Symbionts ; Wood ; Wood - parasitology ; Xylans - metabolism
    ISSN: 2041-1723
    E-ISSN: 2041-1723
    Source: Nature Open Access
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 4
    Language: English
    In: Science (American Association for the Advancement of Science), 2015-07-17, Vol.349 (6245), p.309-312
    Description: Morphinan alkaloids from the opium poppy are used for pain relief. The direction of metabolites to morphinan biosynthesis requires isomerization of (S)- to (R)-reticuline. Characterization of high-reticuline poppy mutants revealed a genetic locus, designated STORR [(S)- to (R)-reticuline] that encodes both cytochrome P450 and oxidoreductase modules, the latter belonging to the aldo-keto reductase family. Metabolite analysis of mutant alleles and heterologous expression demonstrate that the P450 module is responsible for the conversion of (S)-reticuline to 1,2-dehydroreticuline, whereas the oxidoreductase module converts 1,2-dehydroreticuline to (R)-reticuline rather than functioning as a P450 redox partner. Proteomic analysis confirmed that these two modules are contained on a single polypeptide in vivo. This modular assembly implies a selection pressure favoring substrate channeling. The fusion protein STORR may enable microbial-based morphinan production.
    Subject(s): Biosynthesis ; Enzymes ; Metabolites ; Modules ; Morphine ; Narcotics ; Pathways ; Poppies ; Proteins ; REPORTS
    ISSN: 0036-8075
    E-ISSN: 1095-9203
    Source: JSTOR Life Sciences
    Source: Academic Search Ultimate
    Source: Alma/SFX Local Collection
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  • 5
    Language: English
    In: ChemCatChem, 2017-12-20, Vol.9 (24), p.4475-4479
    Description: Chiral secondary and tertiary amines are ubiquitous in pharmaceutical, fine, and specialty chemicals, but their synthesis typically suffers from significant sustainability and selectivity challenges. Biocatalytic alternatives, such as enzyme‐catalyzed reductive amination, offer several advantages over traditional chemistry, but industrial applicability has not yet been demonstrated. Herein, we report the use of cell lysates expressing imine reductases operating at 1:1 stoichiometry for a variety of amines and carbonyls. A collection of biocatalysts with diversity in coverage of small molecules and direct industrial applicability is presented. Biocatalytic reductive amination made easy: Chiral secondary and tertiary amines are ubiquitous in pharmaceutical, fine, and specialty chemicals, but their synthesis typically suffers from significant sustainability and selectivity challenges. Herein, a panel of imine reductases (IREDs) give broad substrate coverage by using stoichiometric reactants with excellent selectivity.
    Subject(s): amination ; Amines ; asymmetric synthesis ; biocatalysis ; Carbonyls ; Catalysis ; Chemical industry ; Enzymes ; Organic compounds ; Reductases ; Synthesis
    ISSN: 1867-3880
    E-ISSN: 1867-3899
    Source: Academic Search Ultimate
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  • 6
    Language: English
    In: Proceedings of the National Academy of Sciences - PNAS, 2013-06-18, Vol.110 (25), p.10189-10194
    Description: Nature uses a diversity of glycoside hydrolase (GH) enzymes to convert polysaccharides to sugars. As lignocellulosic biomass deconstruction for biofuel production remains costly, natural GH diversity offers a starting point for developing industrial enzymes, and fungal GH family 7 (GH7) cellobiohydrolases, in particular, provide significant hydrolytic potential in industrial mixtures. Recently, GH7 enzymes have been found in other kingdoms of life besides fungi, including in animals and protists. Here, we describe the in vivo spatial expression distribution, properties, and structure of a unique endogenous GH7 cellulase from an animal, the marine wood borer Limnoria quadripunctata (LqCel7B). RT-quantitative PCR and Western blot studies show that LqCel7B is expressed in the hepatopancreas and secreted into the gut for wood degradation. We produced recombinant LqCel7B, with which we demonstrate that LqCel7B is a cellobiohydrolase and obtained four high-resolution crystal structures. Based on a crystallographic and computational comparison of LqCel7B to the well-characterized Hypocrea jecorina GH7 cellobiohydrolase, LqCel7B exhibits an extended substrate-binding motif at the tunnel entrance, which may aid in substrate acquisition and processivity. Interestingly, LqCel7B exhibits striking surface charges relative to fungal GH7 enzymes, which likely results from evolution in marine environments. We demonstrate that LqCel7B stability and activity remain unchanged, or increase at high salt concentration, and that the L. quadripunctata GH mixture generally contains cellulolytic enzymes with highly acidic surface charge compared with enzymes derived from terrestrial microbes. Overall, this study suggests that marine cellulases offer significant potential for utilization in high-solids industrial biomass conversion processes.
    Subject(s): 09 BIOMASS FUELS ; Animals ; BASIC BIOLOGICAL SCIENCES ; Binding sites ; biofuel ; Biofuels ; Biological Sciences ; Biomass ; Borers (Insects) ; carbohydrate degrading enzymes ; cellobiohydrolase ; Cellulase - metabolism ; Cellulose ; Cellulose 1,4-beta-Cellobiosidase - chemistry ; Cellulose 1,4-beta-Cellobiosidase - genetics ; Cellulose 1,4-beta-Cellobiosidase - metabolism ; Crustacea - enzymology ; Crustacea - genetics ; Crustaceans ; Crystal structure ; Crystallography, X-Ray ; Crystals ; Digestive System - enzymology ; Enzyme Activation - physiology ; Enzymes ; Fungi ; Gene expression ; gribble ; Health aspects ; Hydrolysis ; Hypocrea - enzymology ; Ligands ; marine animal ; Molecular Sequence Data ; Physiological aspects ; Polymerase chain reaction ; Polysaccharides ; Protein Structure, Tertiary ; Research ; Salt-Tolerance - physiology ; Seawater ; Structure-Activity Relationship ; Substrate Specificity ; Sugars ; Toleration ; Tunnels
    ISSN: 0027-8424
    E-ISSN: 1091-6490
    Source: JSTOR Life Sciences
    Source: HighWire Press (Free Journals)
    Source: Hellenic Academic Libraries Link
    Source: PubMed Central
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  • 7
    Language: English
    In: Revista Brasileira de terapia intensiva, 2021, Vol.33 (2)
    ISSN: 0103-507X
    Source: Academic Search Ultimate
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 8
    Language: English
    In: Applied biochemistry and biotechnology, 2009-07-11, Vol.160 (7), p.1933-1946
    Description: The chit 1 gene from the entomopathogenic fungus Metarhizium anisopliae , encoding the endochitinase CHIT42, was placed under the control of the CaMV 35S promoter, and the resulting construct was transferred to tobacco. Seventeen kanamycin-resistant transgenic lines were recovered, and the presence of the transgene was confirmed by polymerase chain reactions and Southern blot hybridization. The number of chit 1 copies was determined to be varying from one to four. Copy number had observable effects neither on plant growth nor development. Substantial heterogeneity concerning production of the recombinant chitinase, and both general and specific chitinolytic activities were detected in leaf extracts from primary transformants. The highest chitinase activities were found in plants harboring two copies of chit 1 inserts at different loci. Progeny derived from self-pollination of the primary transgenics revealed a stable inheritance pattern, with transgene segregation following a mendelian dihybrid ratio. Two selected plants expressing high levels of CHIT42 were consistently resistant to the soilborne pathogen Rhizoctonia solani , suggesting a direct relationship between enzyme activity and reduction of foliar area affected by fungal lesions. To date, this is the first report of resistance to fungal attack in plants mediated by a recombinant chitinase from an entomopathogenic and acaricide fungus.
    Subject(s): Article ; Biochemistry ; Biological and medical sciences ; Biotechnology ; Blotting, Southern ; Chemistry ; Chemistry and Materials Science ; Chitinases - genetics ; Chitinases - metabolism ; Enzyme Activation - genetics ; Enzymes ; Flowers & plants ; Fundamental and applied biological sciences. Psychology ; Fungi ; Gene Dosage ; Gene expression ; Gene Expression Profiling ; general ; Metarhizium - enzymology ; Metarhizium - genetics ; Plant Diseases - genetics ; Plant Diseases - immunology ; Plant Diseases - microbiology ; Plant Leaves - genetics ; Plant Leaves - immunology ; Plant Leaves - microbiology ; Plants, Genetically Modified ; Polymerase Chain Reaction ; Recombinant Proteins - genetics ; Recombinant Proteins - immunology ; Recombinant Proteins - metabolism ; Rhizoctonia - immunology ; Rhizoctonia - pathogenicity ; Rhizoctonia - physiology ; Tobacco ; Tobacco - genetics ; Tobacco - immunology ; Tobacco - microbiology ; Tobacco industry ; Transgenes
    ISSN: 0273-2289
    E-ISSN: 1559-0291
    Source: Alma/SFX Local Collection
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  • 9
    Language: English
    In: Arquivos brasileiros de cardiologia, 2006-02-01, Vol.86 (2), p.97-104
    Description: OBJETIVO: Avaliar a efetividade do Balão Intra-Aórtico (BIAo) profilático em cirurgia de revascularização miocárdica (CRM) eletiva, para prevenir o infarto trans ou pós-operatório e para reduzir a mortalidade intra-hospitalar nos pacientes com baixa fração de ejeção ventricular esquerda. MÉTODOS: Em modelo de estudo de coorte, foram analisados 239 pacientes com fração de ejeção do ventrículo esquerdo inferior ou igual a 40%, submetidos à CRM eletiva com circulação extracorpórea (CEC), no período compreendido entre março de 1995 a fevereiro de 2001. RESULTADOS: Destes, 58 pacientes receberam BIAo pré-operatório e os demais foram operados sem assistência circulatória (grupo controle). Os dois grupos de pacientes tinham características semelhantes quanto a fatores associados aos desfechos em questão. Ocorreram 5 (8,6%) óbitos no grupo com BIAo e 21 (11,6%), no grupo controle (diferença não-signifícativa). Ocorreram 2 (3,4%) infartos no grupo com BIAo e 28 (15,5%), no grupo controle BIAo (p 〈 0,05), risco relativo de 0,22 com intervalo de confiança de 95% de 0,05 a 0,85. CONCLUSÃO: O BIAo, em uso pré-operatório, pode reduzir de forma significativa o risco de infarto agudo do miocárdio (IAM) trans ou pós-operatório, em pacientes com função sistólica diminuída, sem incremento de complicações vasculares. Nesta mesma situação, o BIAo não diminui de forma significativa a mortalidade. Estudos randomizados devem ser conduzidos para se ter conclusões mais precisas.OBJECTIVE: To evaluate the effectiveness of prophylactic intra-aortic balloon (IAB) in elective myocardial revascularization surgery (MRS), to prevent trans or post-operative infarction and reduce intra-hospital mortality in patients with low left ventricular ejection fraction. METHODS: Using a cohort study model, 239 patients with left ventricular ejection fraction 〈40%, submitted to elective MRS with extracorporeal circulation (ECC) were evaluated from March 1995 to February 2001. RESULTS: Of these, 58 patients received preoperative IAB and the remainder underwent surgery without circulatory assistance (control group). The two groups of patients had similar characteristics regarding factors associated to the pertaining outcomes. There were five demises (8.6%) in the group with IAB and 21 (11.6%) in the control group (non-significant difference). There were 2 (3.4%) infarctions in the IAB group and 28 (15.5%) in the control group (p〈 0.05), relative risk of 0.22 with an interval of confidence of 95% from 0.05 to 0.85 CONCLUSION: The use of pre-operative IAB can significantly reduce the risk of trans or post-operative acute myocardial infarction (AMI) in patients with decreased systolic function, without increasing vascular complications. In this same situation, the IAB does not significantly decrease mortality. Randomized studies are necessary to establish more precise conclusions.
    Subject(s): balão intra-aórtico ; disfunção ventricular ; intra-aortic balloon ; myocardial revascularization ; revascularização miocárdica ; ventricular dysfunction
    ISSN: 0066-782X
    E-ISSN: 1678-4170
    Source: Alma/SFX Local Collection
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 10
    Language: English
    In: Neurology, 2017-10-10, Vol.89 (15), p.1553-1560
    Description: OBJECTIVE:To investigate the temporal pattern and relevant associations of CSF inflammatory measures after intraventricular hemorrhage (IVH). METHODS:We analyzed prospectively collected CSF cell counts and protein and glucose levels from participants in the Clot Lysis Evaluation of Accelerated Resolution of IVH phase III (CLEAR III) trial. Corrected leukocyte count and cell index were calculated to adjust for CSF leukocytes attributable to circulating blood. Data were chronologically plotted. CSF inflammatory measures (daily, mean, median, maximum, and cases with highest quartile response) were correlated with initial IVH volume, IVH clearance rate, thrombolytic treatment, bacterial infection, and adjudicated clinical outcome at 30 and 180 days. RESULTS:A total of 11,376 data points of CSF results from 464 trial participants were analyzed. Measures of CSF inflammatory response evolved during the resolution of IVH. This was significantly more pronounced with initial IVH volume exceeding 20 mL. Intraventricular alteplase was associated with a significantly augmented inflammatory response compared to saline, even after correcting for initial IVH volume. There was an association but nonpredictive correlation of CSF inflammation measures with culture-positive CSF bacterial infection. None of the CSF inflammatory measures, including cases with upper quartile inflammatory response, was associated with a significant detrimental effect on 30 or 180 days functional outcome or mortality after multivariate adjustment for measures of disease severity. CONCLUSIONS:Aseptic CSF inflammation after IVH is primarily dependent on the volume of initial bleed. Thrombolysis intensifies the inflammatory response, with no apparent detrimental effect on clinical outcome. CLINICALTRIALS.GOV IDENTIFIER:NCT00784134.
    Subject(s): 23 ; 7 ; Cerebral Hemorrhage - cerebrospinal fluid ; Cerebral Hemorrhage - diagnostic imaging ; Cerebral Hemorrhage - drug therapy ; Cerebral Hemorrhage - pathology ; Cerebral Ventricles - pathology ; Cytokines - cerebrospinal fluid ; Erythrocytes ; Female ; Fibrinolytic Agents - therapeutic use ; Glucose - metabolism ; Humans ; Leukocyte Count ; Leukocytes - pathology ; Male ; Time Factors ; Tomography Scanners, X-Ray Computed
    ISSN: 0028-3878
    E-ISSN: 1526-632X
    Source: Hellenic Academic Libraries Link
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