Production of an extracellular keratinase from Chryseobacterium sp. growing on raw feathers

The strain Chryseobacterium sp. kr6 shown to be useful for biotechnological purposes such as hydrolysis of poultry feathers and de-hairing of bovine pelts. The effect of temperature, initial pH and media composition on protease production by this keratinolytic strain was studied. The enzyme was produced between 25 and 37\ub0C, with maximum activity and yield at 30\ub0C. When protease production was tested in media with different initial pH, maximum activity was observed when cultivation was carried out at 30\ub0C and initial pH ranging from 6.0 to 8.0. Higher activity was observed when feathers or feather meal were used as growth substrates, followed by soybean meal. The addition of carbohydrates or surfactants to feather broth resulted in decrease in keratinolytic activity

Streaming Motions Towards the Supermassive Black Hole in NGC 1097

We have used GMOS-IFU and high resolution HST-ACS observations to map, in unprecedented detail, the gas velocity field and structure within the 0.7 kpc circumnuclear ring of the SBb LINER/Seyfert 1 galaxy NGC 1097. We find clear evidence of radial streaming motions associated with spiral structures leading to the unresolved (<3.5 parsecs) nucleus, which we interpret as part of the fueling chain by which gas is transported to the nuclear starburst and supermassive black hole.Comment: 4 pages, 3 figures using emulateapj. Accepted for publication in Astrophysical Journal Letters. Download high-resolution version from http://www.astro.uu.se/~kambiz/DOC/paper-N1097.pd

Nuclear spirals as feeding channels to the Supermassive Black Hole: the case of the galaxy NGC 6951

We report the discovery of gas streaming motions along nuclear spiral arms towards the LINER nucleus of the galaxy NGC 6951. The observations, obtained using the GMOS integral field spectrograph on the Gemini North telescope, yielded maps of the flux distributions and gas kinematics in the Halpha, [NII]6584 and [SII]6717,31 emission lines of the inner 7x5 arcsec^2 of the galaxy. This region includes a circumnuclear star-forming ring with radius 500pc, a nuclear spiral inside the ring and the LINER nucleus. The kinematics of the ionized gas is dominated by rotation, but subtraction of a kinematic model of a rotating exponential disk reveals deviations from circular rotation within the nuclear ring which can be attributed to (1) streaming motions along the nuclear spiral arms and (2) a bipolar outflow which seems to be associated to a nuclear jet. On the basis of the observed streaming velocities and geometry of the spiral arms we estimate a mass inflow rate of ionized gas of 3x10^(-4) Msun/yr, which is of the order of the accretion rate necessary to power the LINER nucleus of NGC 6951. Similar streaming motions towards the nucleus of another galaxy with LINER nucleus -- NGC 1097 -- have been reported by our group in a previous paper. Taken together, these results support a scenario in which nuclear spirals are channels through which matter is transferred from galactic scales to the nuclear region to feed the supermassive black hole.Comment: 25 pages, 6 eps figures, accepted for publication in Ap

Evaluation of extracellular proteases from Chryseobacterium sp. Kr6 strain and purification and characterization of a keratinolytic metalloprotease

A linhagem queratinolítica Chryseobacterium sp. Kr6 mostrou-se com possibilidade de aplicação em processos envolvendo queratinólise, principalmente na hidrólise de penas de frango e depilação de couro bovino. No presente trabalho avaliou-se o efeito da composição do meio sobre o crescimento e atividade proteolítica deste isolado e uma protease queratinolítica (queratinase) foi purificada e caracterizada. O microrganismo mostrou-se adaptado à utilização de queratina como substrato durante o crescimento, produziu diferentes proteases dependendo do meio utilizado e a maior atividade proteolítica foi atingida quando utilizado meio de cultivo com penas como única fonte de carbono e nitrogênio. A adição de fonte extra de nutrientes resultou em uma parcial repressão catabólica. Uma protease extracelular (Q1) foi purificada cerca de 14 vezes utilizando cromatografia de interação hidrofóbica em Phenyl-Sepharose CL 4B e gel filtração em Superose H12R. Q1 mostrou ser uma proteína monomérica com peso molecular de 64 KDa determinado por SDS-PAGE e pH e temperatura ótimos de 8,5 e 50°C respectivamente. O perfil de inibição indica tratar-se uma metaloprotease e as seqüências internas dos peptídeos resultantes de digestão tríptica mostraram homologia ao sítio ativo e de ligação ao Zn da família M14 (Carboxipeptidase). A atividade proteolítica foi estimulada pela presença de íons Ca2+ e Mg2+ e inibida por Cu2+, Zn2+, Al2+, Hg 2+ e agentes redutores. Q1 apresentou atividade queratinolítica sobre o substrato keratin azure, mas não foi capaz de hidrolisar penas de frango sugerindo a necessidade de outras enzimas durante o processo de degradação de penas. Utilizando os iniciadores degenerados desenhados com base na seqüência dos peptídeos, foi amplificado um fragmento de 470 pb correspondente a uma região do possível gene desta metaloproteína utilizando DNA e cDNA como molde. A seqüência do fragmento pode estar sendo expressa, mas não apresentou similaridade e homologia a proteínas conhecidas e portando, indicativa de uma nova metaloprotease.The strain Chryseobacterium sp. kr6 shown to be useful for biotechnological purposes such as hydrolysis of poultry feathers and de-hairing of bovine pelts. The effect of media composition on the protease production and growth by this strain was studied and a keratinolytic protease (keratinase) was purified and characterized. The strain was adapted to use keratin as substrate to growth, produced different proteases in different media composition and the higher proteolytic activity was reached when used feather as only source of carbon and nitrogen. The addition of sources of nutrients has resulted in partially repressed catabolism. An extracellular protease Q1) was purified 14-fold by chromatography using the hydrophobic interaction Phenyl-Sepharose CL 4B column and gel filtração in Superose 12HR. SDS-PAGE indicated that the Q1 is a monomeric protein with molecular mass of 64 KDa. and optima pH and temperature were 8,5 e 50°C, respectively. The inhibition profile indicates to be a Zn-metalloprotease and analysis of tryptic peptides sequence revealed sequence homology to the conserved active site and Zn binding site, which may characterize keratinase Q1 as a member of M14 metalloprotease family (Carboxipeptidase). The activity was stimulated by of Ca2+ and Mg2+ and inhibited by Cu2+, Zn2+, Al2+, Hg 2+ and reducing agents. Q1 presented keratinolytic activity under substrate keratin azure, but was unable to hydrolyze poultry feather, suggesting the requirement by other enzymes in the feather hydrolysis mechanism. Degenerate primers amplified a 470 bp, corresponding to a probable gene region of this metalloprotein, with DNA and cDNA. The sequence is being expressed but do not showed similarity and homology to known proteins, thus indicating a new metalloprotease

Keratinolytic bacteria isolated from feather waste

The aim of this study was to characterize keratinolytic bacteria isolated from feather waste. Four isolates were selected after growth on solid medium with feather meal as sole carbon and nitrogen source and screened for proteolytic activity on milk agar plates. Three isolates were Gram-negative (belonged to the genera Burkholderia, Chryseobacterium and Pseudomonas) and one was Gram-positive (Microbacterium sp.). These bacteria grew on diverse keratin wastes such as feather meal, raw feathers, chicken nails, hair and wool. Keratinase activity was detected during growth, but the complete degradation of these substrates was not always achieved. The proteolytic character of crude enzymes was assessed using azokeratin and azocasein as substrates. The keratinases were active on both substrates and were similar in keratin hydrolysis when compared with commercially available microbial peptidases. These novel keratinolytic isolates have potential biotechnological use in processes involving keratin hydrolysis

Keratinolytic bacteria isolated from feather waste Bactérias queratinolíticas isoladas de resíduos de penas

The aim of this study was to characterize keratinolytic bacteria isolated from feather waste. Four isolates were selected after growth on solid medium with feather meal as sole carbon and nitrogen source and screened for proteolytic activity on milk agar plates. Three isolates were Gram-negative (belonged to the genera Burkholderia, Chryseobacterium and Pseudomonas) and one was Gram-positive (Microbacterium sp.). These bacteria grew on diverse keratin wastes such as feather meal, raw feathers, chicken nails, hair and wool. Keratinase activity was detected during growth, but the complete degradation of these substrates was not always achieved. The proteolytic character of crude enzymes was assessed using azokeratin and azocasein as substrates. The keratinases were active on both substrates and were similar in keratin hydrolysis when compared with commercially available microbial peptidases. These novel keratinolytic isolates have potential biotechnological use in processes involving keratin hydrolysis.<br>O objetivo deste estudo foi caracterizar bactérias queratinolíticas isoladas resíduos de penas. Quatro isolados foram selecionados após crescimento em meio sólido contendo farinha de pena como única fonte de carbono e nitrogênio e avaliados quanto a atividade proteolítica em placas de ágar leite. Foram identificadas três linhagens Gram-negativas (pertencentes aos gêneros Burkholderia, Chryseobacterium e Pseudomonas) e uma Gram-positiva (Microbacterium sp.). Estas bactérias cresceram em vários resíduos queratinosos como farinha de pena, penas de frango, unhas de frango, pelos e lã. A atividade queratinolítica foi observada durante crescimento, mas a degradação completa dos substratos não foi observada em todos os casos. O caráter proteolítico das enzimas foi determinado usando azoqueratina e azocaseína como substratos. As queratinases foram ativas em ambos substratos e apresentaram hidrólise de queratina comparável a peptidases microbianas disponíveis comercialmente. Estes novos isolados queratinolíticos apresentam potencial uso biotecnológico em processos relacionados com hidrólise de queratina

Production of an extracellular keratinase from Chryseobacterium sp. growing on raw feathers

The strain Chryseobacterium sp. kr6 shown to be useful for biotechnological purposes such as hydrolysis of poultry feathers and de-hairing of bovine pelts. The effect of temperature, initial pH and media composition on protease production by this keratinolytic strain was studied. The enzyme was produced between 25 and 37°C, with maximum activity and yield at 30°C. When protease production was tested in media with different initial pH, maximum activity was observed when cultivation was carried out at 30°C and initial pH ranging from 6.0 to 8.0. Higher activity was observed when feathers or feather meal were used as growth substrates, followed by soybean meal. The addition of carbohydrates or surfactants to feather broth resulted in decrease in keratinolytic activity

Keratinolytic bacteria isolated from feather waste

The aim of this study was to characterize keratinolytic bacteria isolated from feather waste. Four isolates were selected after growth on solid medium with feather meal as sole carbon and nitrogen source and screened for proteolytic activity on milk agar plates. Three isolates were Gram-negative (belonged to the genera Burkholderia, Chryseobacterium and Pseudomonas) and one was Gram-positive (Microbacterium sp.). These bacteria grew on diverse keratin wastes such as feather meal, raw feathers, chicken nails, hair and wool. Keratinase activity was detected during growth, but the complete degradation of these substrates was not always achieved. The proteolytic character of crude enzymes was assessed using azokeratin and azocasein as substrates. The keratinases were active on both substrates and were similar in keratin hydrolysis when compared with commercially available microbial peptidases. These novel keratinolytic isolates have potential biotechnological use in processes involving keratin hydrolysis