Production and single step purification of cyclodextrin glycosyltransferase from alkalophilic Bacillus firmus by ion exchange chromatography

Savergave, Laxman; Dhule, Santosh S.; Jogdand, V. V.; Nene, Sanjay; Gadre, R. V.

doi:10.1016/j.bej.2007.09.020

Cited by 36 publications

(21 citation statements)

References 24 publications

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“…A CGTase catalisa as reações de transglicosilação intramolecular (ciclização) e intermolecular (ligação, desproporcionamento), bem como apresenta atividade hidrolítica sobre amido e ciclodextrinas ( VAN DER VEEN, et al, 2000a;JOGDAND, 2008) . A reação de transglicosilação da CGTase é operada pelo mecanismo "PingPong", fato verificado por NAKAMURA, HAGA e YAMANAKE (1994).…”

Section: Introductionunclassified

Obtenção De Ciclomaltodextrina-Glucano-Transferase Em Processo Fermentativo Por Bacillus Firmus Cepa 37 Para Produção De Ciclodextrinas.

Bueno¹,

Cavalcanti²,

Melo³

et al. 2015

Anais Do XX Congresso Brasileiro De Engenharia Química

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RESUMO -Com o objetivo exploratório de encontrar melhores condições de produção da enzima Ciclomaltodextrina-Glucano-Transferase (GTase) para formação de Ciclodextrinas, realizou-se cultivo do micro-organismo Bacillus firmus CEPA 37, em batelada utilizando o biorreator BioFlo III, perfazendo um total de 124 horas de fermentação. Para análise de açúcares redutores, as amostras foram submetidas a um processo prévio de hidrólise ácida, com posterior neutralização e reação com DNS (ácido 2,5-dinitrosalicílico), procedendo-se a leitura espectrofotométrica a 600 nm. Para a medida do teor de proteínas solúveis, utilizou-se o método colorimétrico de Bradford. A atividade enzimática foi determinada pelo método das velocidades iniciais envolvendo a complexação da beta-ciclodextrina, produzida pela enzima CGTase com a fenolftaleína e também pela complexação da gama-ciclodextrina com o corante de verde de bromocresol. A Atividade específica é a razão entre a atividade enzimática e a concentração de proteínas solúveis no meio. Para este experimento, a máxima atividade específica obtida em relação a beta-CD foi de 1,8 U/mg de proteína em 120 horas de cultivo, e para gama-CD, obteve-se 0,24 U/mg de proteína em 103 horas de cultivo. INTRODUÇÃOA degradação enzimática do amido geralmente resulta na produção de glicose, maltose, maltotriose, etc., isto é, maltooligômeros de cadeias lineares ou ramificadas, conhecidos como dextrinas. Esse tipo de degradação do amido é um processo de hidrólise em que o produto resultante provém da quebra da ligação glicosídica acompanhada da adição de uma molécula de água, contudo, se o amido é degradado por uma enzima do tipo glicosiltransferase (CGTase), o produto resultante da quebra da cadeia é submetido a uma reação intramolecular sem a participação da molécula de água. Produtos cíclicos de ligação alfa-1,4 são formados se a enzima for uma ciclomaltodextrina glucanotransferase, e são conhecidos como ciclodextrinas (FRÖMMING;SZEJTLI, 1994;DEL VALLE, 2004; COSTA et al., 2007). As ciclodextrinas (CDs) são oligossacarídeos cíclicos não redutores compostos principalmente por 6, 7 e 8 unidades de glicose, as quais são denominadas de alfa, beta e gama-CDs, respectivamente (TONKOVA 1998; CEREDA et al., 2004;FERREIRA et al., Área temática: Processos Biotecnológicos 1

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Section: Introductionunclassified

Obtenção De Ciclomaltodextrina-Glucano-Transferase Em Processo Fermentativo Por Bacillus Firmus Cepa 37 Para Produção De Ciclodextrinas.

Bueno¹,

Cavalcanti²,

Melo³

et al. 2015

Anais Do XX Congresso Brasileiro De Engenharia Química

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“…All were screened. Most concerned plant protection and nematicidale activities (Castillo et al, 2011;Crow, 2010;Hafez and Sundararaj 2009;Mendoza et al, 2008;Mendoza and Sikora 2009;Schrimsher et al, 2011;Terefe et al, 2009), polysaccharides and lipids biotechnology (Deepthi et al, 2011;Elayaraja et al, 2011;Jaitak et al, 2009;Jommuengbout et al, 2009;Kumar and Kannan, 2011;Mazzer et al, 2008;Pazzetto et al, 2011;Savergave et al, 2008;Sornyotha et al 2010), production of various enzymes, water and waste water treatment (Arora et al 2011;Sau et al, 2008;Kumar et al, 2010), use of dead bacterial cells as vaccines adjuvant (Havlíčková et al, 2011;Zanvit et al, 2010). No article concerned the presence of Bacillus firmus in the food or feed chain and none permitted to assess its innocuity for human and animals.…”

Section: Bacillus Firmusmentioning

confidence: 99%

Scientific Opinion on the maintenance of the list of QPS biological agents intentionally added to food and feed (2012 update)

2012

EFS2

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EFSA is requested to assess the safety of a broad range of biological agents in the context of notifications for market authorisation as sources of food and feed additives, enzymes and plant protection products. The qualified presumption of safety (QPS) assessment was developed for safety risk assessments to provide a harmonised generic pre-assessment to support EFSA"s scientific Panels. The safety of unambiguously defined biological agents at the highest taxonomic unit appropriate for the purpose for which an application is intended and the completeness of the body of knowledge are assessed. Identified safety concerns for a taxonomic unit are where possible and reasonable in number reflected as "qualifications" with a recommendation for the QPS list. The list of QPS recommended biological agents is reviewed and updated annually. Therefore, the only valid list is the one in the most recently published scientific opinion. The 2012 update reviews microorganisms previously assessed including bacteria, yeasts, filamentous fungi and viruses used for plant protection purposes. The BIOHAZ Panel confirmed all taxonomic units previously recommended for the QPS list. The notifications were reviewed. Bacillus firmus was re-evaluated and not recommended for the QPS list. A new recommendation was made for Leuconostoc pseudomesenteroides. Carnobacterium maltaromaticum was assessed for the first time and not recommended for the QPS list. Qualifications for the taxonomic units included in the QPS recommended list were reviewed and confirmed. Filamentous fungi and enterococci were not recommended for the QPS list following updating and reviewing of current scientific knowledge. For Enterococcus faecium recent data indicate a possible distinction between pathogenic and non-pathogenic strains. This is considered too recent knowledge for a QPS recommendation, considering the recent information on the evolution of the epidemiology of Enterococcus infections in humans. Tasks three required, for the taxonomic units included in the QPS recommended list, a review and update of knowledge concerning antimicrobial resistance and a review of the qualifications. The information of the previous opinion was updated by the BIOHAZ Panel and the qualifications were confirmed.The final aspect included a review of the body of knowledge for filamentous fungi and enterococci. The BIOHAZ Panel updated the knowledge of filamentous fungi notified to EFSA. Although numerous data, published since the 2011 QPS opinion, have contributed to partially fulfil gaps of knowledge, too many unknowns remain in 2012 to allow a filamentous fungus to be recommended for the QPS list.Enterococcus faecium is not recommended for the QPS list in spite of the recent scientific knowledge allowing a differentiation of pathogenic from non-pathogenic strains. This is of value for the FEEDAP Scientific Panel dealing with the strain specific notification, but it is too recent knowledge for a QPS recommendation, considering the recent information on the evolution of the ep...

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“…Cyclization activity was measured, according to a modified method by Savergave et al (2008), as a function of the β-CD production rate, using soluble starch as substrate at 1% (w/v) in 50 mM Tris-HCl buffer, pH 7.0, at 65°C for 15 min. The β-CD produced in the assay was determined based on colour fading at 550 nm.…”

Section: Cyclization Activitymentioning

confidence: 99%

Production of a novel Cyclodextrin glycosyltransferase from Bacillus sp. SK13.002

Sun¹,

Letsididi²,

Pan³

2013

Afr. J. Microbiol. Res.

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A β-cyclodextrin glycosyltransferase (β-CGTase) from a new alkalitolerant Bacillus sp. SK 13.002 strain which can significantly hydrolyze starch into short linear saccharides, in addition to production of cyclodextrins (CDs) was produced. The hydrolytic activity of this CGTase as a side reaction is thought to be due to partial retention of ancestral enzyme function from evolution over time. This CGTase is therefore another example of an enzyme at an intermediary stage in between ''true'' α-amylases and ''true'' CGTases. The strain also produced two CGTase isozymes which were purified to homogeneity using DEAE-Sepharose anion exchange chromatography and Superdex 75 gel filtration chromatography to show Molecular masses of 67.5 and 46.8 kDa on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) respectively, which were confirmed by liquid chromatography/mass spectrometry (LC/MS) analysis to be 67.6 and 47.3 kDa, respectively. Both CGTase isozymes formed CDs from soluble starch. Most of the reported CGTases were composed of one single enzyme with only a few strains having CGTase isozymes showing different isoelectric points. Therefore, Bacillus sp. SK 13.002 strain is another Bacillus capable of producing CGTase isozymes which were individually purified to homogeneity.

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Production and single step purification of cyclodextrin glycosyltransferase from alkalophilic Bacillus firmus by ion exchange chromatography

Cited by 36 publications

References 24 publications

Obtenção De Ciclomaltodextrina-Glucano-Transferase Em Processo Fermentativo Por Bacillus Firmus Cepa 37 Para Produção De Ciclodextrinas.

Obtenção De Ciclomaltodextrina-Glucano-Transferase Em Processo Fermentativo Por Bacillus Firmus Cepa 37 Para Produção De Ciclodextrinas.

Scientific Opinion on the maintenance of the list of QPS biological agents intentionally added to food and feed (2012 update)

Production of a novel Cyclodextrin glycosyltransferase from Bacillus sp. SK13.002

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