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Adhesion and production of degrading enzymes by bacteria isolated from biofilms in raw milk cooling tanks Ciênc. Tecnol. Aliment.
Flach,Juliana; Grzybowski,Valeska; Toniazzo,Geciane; Corção,Gertrudes.
Biofilms in milk cooling tanks compromise product quality even on farms. Due to the lack of studies of this topic, this study evaluated the microbiological conditions of raw milk cooling tanks on farms and characterized the microorganisms isolated from these tanks. Samples were wiped off with sterile swabs from seven milk cooling tanks in three different points in each tank. Mesophiles and psychrotrophic counts were performed in all samples. The isolation of Pseudomonas spp., Bacillus cereus and atypical colonies formed on selective media were also performed, totalizing 297 isolates. All isolates were tested for protease and lipase production and biofilm formation. Of the total isolates, 62.9% produced protease, 55.9% produced lipase, and 50.2% produced...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Psychrotrophic bacteria; Cleaning process; Lipase; Protease.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612014000300020
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Analysis of castor bean ribosome-inactivating proteins and their gene expression during seed development Genet. Mol. Biol.
Loss-Morais,Guilherme; Turchetto-Zolet,Andreia Carina; Etges,Matheus; Cagliari,Alexandro; Körbes,Ana Paula; Maraschin,Felipe dos Santos; Margis-Pinheiro,Márcia; Margis,Rogério.
Ribosome-inactivating proteins (RIPs) are enzymes that inhibit protein synthesis after depurination of a specific adenine in rRNA. The RIP family members are classified as type I RIPs that contain an RNA-N-glycosidase domain and type II RIPs that contain a lectin domain (B chain) in addition to the glycosidase domain (A chain). In this work, we identified 30 new plant RIPs and characterized 18 Ricinus communis RIPs. Phylogenetic and functional divergence analyses indicated that the emergence of type I and II RIPs probably occurred before the monocot/eudicot split. We also report the expression profiles of 18 castor bean genes, including those for ricin and agglutinin, in five seed stages as assessed by quantitative PCR. Ricin and agglutinin were the most...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Agglutinin; Evolution; Lipase; Ricinus communis; Ricin; RIPs.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572013000100011
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Aplicação de lipase e monoglicerídeo em pão de forma enriquecido com fibras Ciênc. Tecnol. Aliment.
Gandra,Kelly Moreira; Del Bianchi,Michelle; Godoy,Vanessa Padovani; Queiroz,Fernanda Paula Collares; Steel,Caroline Joy.
Neste trabalho, estudou-se a aplicação da enzima lipase e do emulsificante monoglicerídeo em pão de forma enriquecido com fibras, com o objetivo de verificar a possibilidade de substituição do emulsificante pela enzima. Inicialmente, foi realizada a caracterização das matérias-primas principais (farinha e farelo de trigo). Os pães de forma foram elaborados pelo método de massa direta. Foi utilizado um planejamento experimental do tipo composto central rotacional com duas variáveis independentes: i) dosagem de lipase; e ii) dosagem de monoglicerídeo e, paralelamente, realizou-se um teste controle (sem adição de lipase e monoglicerídeo) para comparação. As variáveis dependentes foram as características de qualidade dos pães: i) volume específico; ii)...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Pão de forma; Lipase; Monoglicerídeo; Fibras.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612008000100027
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Aproveitamento de resíduos da agroindústria do óleo de dendê para a produção de lipase por Aspergillus Níger Ciência Rural
Penha,Edmar das Mercês; Viana,Ludmila de Araújo Nunes; Gottschalk,Leda Maria Fortes; Terzi,Selma da Costa; Souza,Erika Fraga de; Freitas,Sidinéa Cordeiro de; Santos,Juliana de Oliveira; Salum,Thaís Fabiana Chan.
RESUMO: Neste trabalho, foi avaliado o desempenho de duas linhagens de Aspergillus niger (mutante 11T53A14 e selvagem C) previamente selecionadas como promissoras para a produção de lipases, utilizando meios de cultivo formulados a partir da torta de dendê (palmiste) e da borra alcalina do refino do óleo de dendê (borra de dendê), resíduos provenientes da agroindústria do óleo de dendê (palma) por fermentação em estado sólido (FES). Os experimentos de produção da enzima em FES foram conduzidos em colunas aeradas, incubadas em banho-maria a 32ºC com entrada controlada de ar não umedecido de 1,0vvm. Os melhores resultados foram obtidos com a cepa mutante A. niger 11T53A14 em meio contendo torta de dendê umedecida com solução sulfato de amônio (1,2%) e com a...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Lipase; Resíduo agroindustrial; Aspergillus niger.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016000400755
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Aqueous extract of Psidium guajava leaves: phenolic compounds and inhibitory potential on digestive enzymes Anais da ABC (AABC)
SIMÃO,ANDERSON A.; MARQUES,TAMARA R.; MARCUSSI,SILVANA; CORRÊA,ANGELITA D..
ABSTRACT Leaves of Psidium guajava L. (guava) have been widely used in the popular way for prevention and treatment of various diseases. Thus, the objective of this study was to evaluate the inhibitory potential of leaves aqueous extract from three cultivars of P. guajava (Pedro Sato, Paluma and Século XXI) on α-amylase, α-glycosidase, lipase, and trypsin enzymes, in the presence or not of simulated gastric fluid and to determine the content of phenolic compounds by high performance liquid chromatography. All cultivars presented the same composition in phenolic compounds, but in different proportions. The compounds identified are gallic acid, epigallocatechin gallate, syringic acid, o-coumaric acid, resveratrol, quercetin, and catechin (which was the major...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Medicinal plants; Obesity; Α-amylase; Α-glycosidase; Lipase; Trypsin.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652017000502155
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Assessment of lipolytic activity of isolated microorganisms from the savannah of the Tocantins Biological Sciences
Soares, Marysa de Kássia Guedes; Facundes, Bruna Coelho; Chagas Júnior, Aloísio Freitas; Silva, Ezequiel Marcelino da.
Current study assesses the biodiversity and selects lipase-producer microorganisms with industrial interest, from the savannah of the state of Tocantins, Brazil. Seventeen pequi microorganisms (Caryocar brasiliense) were isolated in the decomposition stage and 35 microorganisms were retrieved from the soil fraction under the collected pequi. Yarrowia lypolitica strain was used as positive control in all assays. The 52 strains were subjected to tests in a solid medium with Tween 20 for checking halos formed by crystals, indicating lipase production by inoculated strains. Another test to confirm lipase producers was conducted in microplates with liquid medium and enriched with p-nitrophenyl palmitate (pNPP) monitored at 410 ηm. The AS16 and AP5 strains...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Lipase; Caryocar brasiliense; Yarrowia lypolitica; P-nitropheny lpalmitate.
Ano: 2015 URL: http://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/24324
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Atividade enzimática de isolados de rizóbia nativos da Amazônia Central crescendo em diferentes níveis de acidez Ciênc. Tecnol. Aliment.
Oliveira,Arlem Nascimento de; Oliveira,Luiz Antonio de; Andrade,Jerusa Souza; Chagas Júnior,Aloisio Freitas.
A importância das bactérias conhecidas como rizóbia no estabelecimento de leguminosas tem sido amplamente reconhecida. Porém, poucas são as informações referentes ao perfil enzimático dessas bactérias benéficas. O estudo objetivou investigar a influência do pH do meio sólido sobre a atividade enzimática de rizóbios nativos da Amazônia Central. Essa triagem constitui o primeiro passo no processo de seleção de microorganismos benéficos, como produtores de enzimas de aplicação biotecnológica. Nesse estudo, 64 isolados de rizóbia foram testados para a produção extracelular de amilase, lipase, pectinase e protease, em meio YMA modificado. Excetuando a atividade pectinolítica, todas as outras enzimas (amilase, lipase e protease) foram detectadas nos isolados...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Amilase; Lipase; Pectinase; Protease; Feijão caupi; Soja.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612006000100032
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Atividade lipolítica do leite com células somáticas ajustadas para diferentes níveis Arq. Bras. Med. Vet. Zootec.
Santos,M.V.; Oliveira,C.A.F.; Augusto,L.F.B.; Aquino,A.A..
Estimou-se a origem da atividade lipolítica do leite com contagens de células somáticas (CCS) ajustadas para diferentes níveis. A partir de lotes de leite cru com baixa (100.000 cel/ml) e alta (1.000.000 cel/ml) CCS, formaram-se três tratamentos: A) leite com baixa CCS; B) leite com alta CCS; C) leite originalmente com baixa CCS e com adição de células somáticas. O leite foi submetido à pasteurização e armazenado por 21 dias sob refrigeração a 6ºC. Durante o período de armazenamento, foram coletadas amostras nos dias 1, 7, 14 e 21 para avaliação da concentração de ácidos graxos livres (AGL). Não foi observado efeito de tratamento sobre a concentração de AGL, durante todo o período de armazenamento e nem em cada dia de coleta isoladamente, cujas médias de...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Leite; Células somáticas; Atividade enzimática; Lipase.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352007000400002
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Characterization of biotechnologically relevant extracellular lipase produced by Aspergillus terreus NCFT 4269.10 BJM
Sethi,Bijay Kumar; Nanda,Prativa Kumari; Sahoo,Santilata.
Abstract Enzyme production by Aspergillus terreus NCFT 4269.10 was studied under liquid static surface and solid-state fermentation using mustard oil cake as a substrate. The maximum lipase biosynthesis was observed after incubation at 30 °C for 96 h. Among the domestic oils tested, the maximum lipase biosynthesis was achieved using palm oil. The crude lipase was purified 2.56-fold to electrophoretic homogeneity, with a yield of 8.44%, and the protein had a molecular weight of 46.3 kDa as determined by SDS-PAGE. Enzyme characterization confirmed that the purified lipase was most active at pH 6.0, temperature of 50 °C, and substrate concentration of 1.5%. The enzyme was thermostable at 60 °C for 1 h, and the optimum enzyme–substrate reaction time was 30...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Aspergillus terreus; Lipase; Liquid static surface fermentation; Solid-state fermentation.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000100143
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Characterization of Lipase from Bacillus subtilisI-4 and Its Potential Use in Oil Contaminated Wastewater BABT
Iqbal,Syeda Abeer; Rehman,Abdul.
ABSTRACTA lipase producing bacterium was isolated from oil contaminated effluents of various industries from Sheikhupura Road, Pakistan, and, on the basis of biochemical and 16S rRNA ribotyping, was identified asBacillus subtilis. The optimum temperature and pH for the growth of the culture were 37ºC and 7.0, respectively.B. subtilis I-4 had a lag phase of 4 h in LB medium while this phase prolonged to 6 h in oil containing medium. The optimum temperature and pH for the enzyme activity were 50ºC and 7.0, respectively. Maximum lipase activity was found in the presence of Ca ions. Olive oil and Tween 80 induced lipase gene in the bacterium while concentration of oil greater than 2% retarded the growth of the organism. In addition to lipaseB. subtilis I-4...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bacillus subtilisI-4; Lipase; Alkane hydroxylase.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000500789
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Citral as food additive for common snook - zootechnical parameters and digestive enzymes Ciência Rural
Michelotti,Bruna Tomazetti; Mori,Natacha Cossettin; Magnotti,Caio Cesar Franca; Heinzmann,Berta Maria; Almeida,Ana Paula Gottlieb; Cerqueira,Vinicius Ronzani; Baldisserotto,Bernardo.
ABSTRACT: Essential oils of plants whose main compound is citral showed beneficial effects when added to fish feed. The objective of the present study was to evaluate the dietary effect of the addition of citral on zootechnical parameters and digestive enzyme activities of Centropomus undecimalis. Juveniles were fed for 45 days with diets containing different amounts of citral (0.0 - control, 0.5, 1.0, and 2.0 mL per kg of diet). The water quality parameters were kept stable during the experiment and no mortality was observed. At the end of the experimental period, the treatment 0.5 mL citral per kg of diet had the lowest weight gain and specific growth rate, and the highest feed conversion, while the same parameters did not differ between the other...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Centropomus undecimalis; Amylase; Pepsin; Lipase; Growth; Marine fish farming.
Ano: 2020 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782020000400651
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Clavulanic acid production by the MMS 150 mutant obtained from wild type Streptomyces clavuligerus ATCC 27064 BJM
Vasconcelos,Eliton da Silva; Lima,Vanderlei Aparecido de; Goto,Leandro Seiji; Cruz-Hernández,Isara Lourdes; Hokka,Carlos Osamu.
Clavulanic acid (CA) is a powerful inhibitor of the beta-lactamases, enzymes produced by bacteria resistants to penicillin and cefalosporin. This molecule is produced industrially by strains of Streptomyces clavuligerus in complex media which carbon and nitrogen resources are supplied by inexpensive compounds still providing high productivity. The genetic production improvement using physical and chemical mutagenic agents is an important strategy in programs of industrial production development of bioactive metabolites. However, parental strains are susceptible to loss of their original productivity due genetic instability phenomenona. In this work, some S. clavuligerus mutant strains obtained by treatment with UV light and with MMS are compared with the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Clavulanic acid; Streptomyces clavuligerus; Mutation; Lipase.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822013000400005
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Comparative esterification of phenylpropanoids versus hydrophenylpropanoids acids catalyzed by lipase in organic solvent media Electron. J. Biotechnol.
Cassani,Julia; Luna,Héctor; Navarro,Arturo; Castillo,Edmundo.
The esterification of phenylpropanoid and hydrophenylpropanoid acids, catalyzed by candida antarctica lipase B (CAL-B), with several alcohols has demonstrated that the substitution pattern on the aromatic ring has a very significant influence on the reactivity of the carboxyl group due, mainly, to electronic effects, when compared to the unsaturated acids with the hydrogenated acids. It is also clear that in the saturated acids there still remain some unclear effects related to the aromatic substituents.
Tipo: Journal article Palavras-chave: CAL-B; Electronic effects; Esterification; Lipase; Phenylpropanoid acids.
Ano: 2007 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000400004
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Composition of gastrointestinal content, protease and lipase activities in summer and winter of four freshwater siluriforms (Teleostei: Actinopterygii) with two different feeding habits Rev. Bras. Zool.
Almeida,Ana Paula Gottlieb; Zardo,Everton Luis; Toni,Candida; Behr,Everton Rodolfo; Silva,Leila Picolli da; Vieira,João Paes; Loro,Vania Lúcia; Baldisserotto,Bernardo.
ABSTRACT The aim of this study was to determine the composition of gastrointestinal content and protease and lipase activities in summer and winter as well as to evaluate the relationship between digestive enzyme activity and centesimal composition of gastrointestinal content and feeding habits of two omnivorous species, Rhamdia quelen (Quoy & Gaimard, 1824) and Pimelodus maculatus (LaCèpede, 1803), and of two detritivorous species, Loricariichthys anus (Valenciennes, 1835) and Hypostomus commersoni (Valenciennes, 1836). The activities of pepsin, trypsin, chymotrypsin, and lipase, and the levels of proteins and lipids in the gastrointestinal tract, were evaluated. The enzyme activities were not related to the centesimal composition of gastrointestinal...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Chymotrypsin; Fish; Lipase; Pepsin; Trypsin.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-46702018000100322
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Concomitant production of protease and lipase by Bacillus licheniformis VSG1: production, purification and characterization BJM
Sangeetha,R.; Geetha,A.; Arulpandi,I..
This study was aimed at producing protease and lipase simultaneously on a common medium by Bacillus licheniformis VSG1, which was isolated from a tannery effluent. The effect of media composition with respect to protein source, lipid source and emulsifier on the production of protease and lipase was analysed. Both those enzymes were produced under optimized conditions like pH, temperature and incubation time. The enzyme mixture comprising of both protease and lipase was purified by ammonium sulphate precipitation, dialysis and gel filtration chromatography to obtain 20-fold pure enzymes. The purified enzyme mixture was characterized to determine the optimum pH and temperature of protease and lipase, the response of the enzymes to inhibitors, additives and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bacillus licheniformis; Protease; Lipase; Concomitant production; Purification.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000100026
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Efeito da suplementação enzimática em rações com farelo de soja e soja integral extrusada sobre a digestibilidade de nutrientes, o fluxo de nutrientes na digesta ileal e o desempenho de frangos R. Bras. Zootec.
Garcia,Elis Regina de Moraes; Murakami,Alice Eiko; Branco,Antonio Ferriani; Furlan,Antonio Claudio; Moreira,Ivan.
Três experimentos foram realizados com o objetivo de avaliar o efeito da suplementação enzimática em rações à base de milho e soja sobre o desempenho (Experimento 1), a digestibilidade de nutrientes (Experimento 2) e o desempenho e o fluxo de nutrientes na digesta ileal (Experimento 3) em frangos de corte. No Experimento 1, para o cálculo das rações experimentais, a composição química do farelo de soja (FS) e da soja integral extrusada (SIE) foi superestimada em 7% para os valores de EM e PB e 5% para Met, Met+Cis e Lis (Trat. C, D e E) e 9% para EM, 7% para PB e 5% para Met, Met+Cis e Lis (Trat. F, G e H), quando houve a adição de enzimas (Allzyme Vegpro<FONT FACE=Symbol>â</FONT> e Allzyme Lipase<FONT FACE=Symbol>â</FONT>)....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Complexo multienzimático; Farelo de soja; Frango de corte; Lipase; Soja integral extrusada.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982000000500020
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Efeitos do período de jejum entre o nascimento e o alojamento de pintos de corte e da adição de óleo à ração sobre o desenvolvimento do trato gastrintestinal e concentração de lipase Arq. Bras. Med. Vet. Zootec.
Cançado,S.V.; Baião,N.C..
Os objetivos deste trabalho foram determinar o efeito de três períodos de jejum (zero, 24 e 48 horas) entre o nascimento e o alojamento de pintos e da adição de óleo à ração sobre o crescimento alométrico dos órgãos digestivos e estudar a atividade da lipase no pâncreas e intestino delgado. Foram utilizados 720 pintos de corte machos, criados de um a 15 dias de alojamento. As avaliações do crescimento dos órgãos digestivos foram feitas nos dias zero, três, seis, nove, 12 e 15 de idade. As avaliações da concentração de lipase no pâncreas foram feitas nos dias zero, três, seis, nove, 12 e 15, e no intestino delgado nos dias zero, três e seis. O delineamento experimental foi em arranjo fatorial 3 x 2 (três períodos de jejum e dois tipos de ração). O...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Pintos de corte; Trato gastrintestinal; Lipase; Jejum.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352002000600011
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Effect of methyl branching of C8H18 alkanes and water activity on lipase-catalyzed enantioselective esterification of ibuprofen Electron. J. Biotechnol.
Gottemukkala,Vijay V; Saripella,Kalyan K; Kadari,Anil K; Neau,Steven H.
The purpose of this research was to study the effect of the methyl branching of a high log P alkane solvent and the water activity in the organic medium on the initial rate and the enantioselectivity of ibuprofen esterification catalyzed by Candida rugosa lipase. Resolution of ibuprofen is important because S-(+)-ibuprofen has the desired pharmacological activity, whereas the R-(-)-enantiomer causes much of the side effects. The Candida rugosa lipase-catalyzed reaction in isooctane at 40ºC and 0.73 water activity gave the best results, both in terms of the initial reaction rate and the enantioselectivity of the reaction. An increase in water activity allowed a higher reaction rate and enantiomeric excess in each of the four solvents. An increase in methyl...
Tipo: Journal article Palavras-chave: Enantioselective; Esterification; Ibuprofen; Lipase; Solvent-effect; Water activity.
Ano: 2008 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100002
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Effects of Lysophospholipid and Lipase Enzyme Supplementation to Low Metabolizable Energy Diets on Growth Performance, Intestinal Morphology and Microbial Population and Some Blood Metabolites in Broiler Chickens Rev. Bras. Ciênc. Avic.
Movagharnejad,M; Kazemi-Fard,M; Rezaei,M; Teimuri-Yansari,A.
ABSTRACT This experiment aimed to evaluate the impacts of dietary lysophospholipid (LPL) and lipase enzyme complementation based on low-energy diets on growth performance, intestinal morphology, blood metabolites, immune response, and carcass traits in broiler chickens. Two hundreds broiler chicks were assigned to a completely randomized design with five treatments and four replications with ten one-day old chicks. The five treatments were: positive control (PC) without LPL supplementation and adequate in all nutrients, negative control (NC) without LPL the reduced 150 kcal/kg of metabolizable energy, NC+ 0.15% LPL (LPL15), NC+ lipase (NCL), NC+ 0.15% LPL+ lipase (NCLL). Feeding LPL improved body weight gain and feed conversion ratio (FCR). In contrast,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Broiler Chicken; Lipase; Metabolizable Energy; Growth Performance; Intestinal Morphology.
Ano: 2020 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2020000200316
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Endophytic fungi found in association with Bacopa monnieri as potential producers of industrial enzymes and antimicrobial bioactive compounds BABT
Katoch,Meenu; Salgotra,Aseem; Singh,Gurpreet.
This study aimed to screen the endophytic fungal species of ethano-medicinal plant Bacopa monnieri (L.) Pennell for their ability to produce antimicrobial substances against Bacillus subtilis, Pseudomonas aeroginosa, Salmonella typhimurium, Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, and Candida albicans. Endophytes were also screened for their ability to produce amylase, cellulase, protease and lipase to evaluate their ecological role within the host plant. Twenty-six endophytes were isolated and seventeen were identified. All the isolated endophytes exhibited amylolytic activity. Lipolytic, cellulolytic, proteolytic activity was shown by 98, 28 and 31% isolates, respectively. Similarly, all the endophytes (100%) exhibited significant...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bacopa monnieri; Antimicrobial; ITS 5.8S rDNA; Lipase; Amylase; Endophyte.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000500714
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