Screening for novel lipolytic enzymes from uncultured soil microorganisms

Lee, Seon-Woo; Won, Keehoon; Lim, He Kyoung; Kim, Jin-Cheol; Choi, Gyung Ja; Cho, Kwang Yun

doi:10.1007/s00253-004-1722-3

Cited by 160 publications

(113 citation statements)

References 39 publications

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“…However, our approach of library preparation in a fosmid and subsequent subcloning was more efficient in searching for esterolytic activity. Fosmids are good vectors for constructing metagenomic libraries because of their high cloning efficiency, improved stability in E. coli, and large insert size [23]. Our results indicate that the esterase activity obtained in this study could be somewhat different from the esterase of previously cultured bacteria.…”

Section: Est1r Activitymentioning

confidence: 53%

“…Recently, novel lipolytic enzymes and genes have been identified from metagenomic libraries of soil [7,23], hot spring sediments [30], pond water [29], and alkaline soda lakes [28]. Lipolytic enzymes, including lipases (EC 3.1.1.3) and esterases or carboxylesterases (EC 3.1.1.1), have been found in a wide range of organisms from bacteria to humans [18,19].…”

Section: Introductionmentioning

confidence: 99%

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Cloning and Characterization of a Novel Carboxylesterase Gene from Cow Rumen Metagenomic Library

Islam¹,

Kim²,

Renukaradhya³

et al. 2010

Journal of Life Science

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The gene encoding esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est1R) was 2,465 bp in length, encoding a protein of 366 amino acid residues, and the molecular weight of the enzyme was 61,166 Da. Est1R of rumen cosmid library shared 5.9% amino acid identity with Est1R (P37967) of PNB carboxylesterase, 6.1% with Est1R (1EEAA) of acetylcholinesterase and 6.1% with Est1R (1H23A) of chain A. BlastP in NCBI database analysis of Est1R revealed that it was not homologous to previous known lipases and esterases. Est1R showed optimum activity at pH 7.0 and 40 o C. On the other hand, the enzyme was found to be most active without organic solvent, followed by 95% activity with methanol, and the enzyme activity was highly affected by hexane (lost 51% activity). Therefore, the novel esterase gene est1R is likely obtainable from cow rumen metagenome and may be utilized for industrial purposes.

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Section: Est1r Activitymentioning

confidence: 53%

Section: Introductionmentioning

confidence: 99%

Cloning and Characterization of a Novel Carboxylesterase Gene from Cow Rumen Metagenomic Library

Islam¹,

Kim²,

Renukaradhya³

et al. 2010

Journal of Life Science

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“…Similarly a number of lipolytic genes or esterases have been isolated. In one such report, out of the 33700 fosmid clones from a metagenomic library prepared from top forest soil, 8 unique lipolytic clones were identifi ed on the basis of clearing zone on LB-tributyrin medium [87]. The subcloning of the lipolytic clones in the small insert library yielded six novel genes that encoded for lipolytic enzymes with 34-48% similarity to known enzymes.…”

Section: Enzyme and Natural Productsmentioning

confidence: 99%

“…The quality of DNA in terms of size, amount and presence of contaminants should be considered before the library is made as poor quality DNA can lead to low coverage of environmental genomes in the library [80]. Extracted DNA is purifi ed if required and cloned in suitable vectors, such as plasmid [84,85], cosmid [86], fosmid [63,87] or BAC [88,89]. As small insert libraries do not permit detection of large gene clusters and operons, large insert libraries in cosmid/ fosmid/ BAC vectors are preferred [80].…”

Section: Metagenomic Librariesmentioning

confidence: 99%

From bacterial genomics to metagenomics: concept, tools and recent advances

et al. 2008

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In the last 20 years, the applications of genomics tools have completely transformed the fi eld of microbial research. This has primarily happened due to revolution in sequencing technologies that have become available today. This review therefore, fi rst describes the discoveries, upgradation and automation of sequencing techniques in a chronological order, followed by a brief discussion on microbial genomics. Some of the recently sequenced bacterial genomes are described to explain how complete genome data is now being used to derive interesting fi ndings. Apart from the genomics of individual microbes, the study of unculturable microbiota from different environments is increasingly gaining importance. The second section is thus dedicated to the concept of metagenomics describing environmental DNA isolation, metagenomic library construction and screening methods to look for novel and potentially important genes, enzymes and biomolecules. It also deals with the pioneering studies in the area of metagenomics that are offering new insights into the previously unappreciated microbial world.

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“…As enzimas que hidrolisam os lipídios são denominadas de lipases e podem ser encontradas em animais, vegetais e micro-organismos (CASTRO et al, 2009;LEE et al, 2004;VADLAMANI;PARCHA, 2011;BRANDELLI, SALA, KALIL, 2015 ( MENDES et al, 2005;GASPARIN et al, 2012).…”

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Avaliação Do Potencial Biotecnológico De Aspergillus Parasiticus Ucp 1281 No Biotratamento De Efluentes Da Indústria De Laticínios E Produção De Lipídeos

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Leão

Souza

et al. 2015

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RESUMO: Neste trabalho foi realizada a identificação de Aspergillus sp (SIS 16), isolado de amostra do solo da região da Caatinga no estado de Pernambuco, e avaliado o potencial biotecnológico deste, no biotratamento de efluentes da indústria láctea. O isolado foi identificado, através das características macroscópicas e microscópicas como A. parasiticus (UCP1281). (MONTEIRO, 2012;LIU et al., 2012;SMRITHI et al., 2013;MONCIARDINI et al., 2014).A Caatinga é um ecossistema do sertão nordestino.Estende-se por cerca de 735.000 km 2 , sendo limitada a leste e a oeste pelas florestas Atlântica e Amazônica, respectivamente, e ao sul pelo Cerrado (REIS, 1976). Apresenta um clima semi-árido, quente e com baixos índices pluviométricos, baixa umidade relativa do ar e altas temperaturas durante quase todo o ano (COSTA et al., 2014 (LIU et al., 2008;NOTARNICOLA et al., 2012; JOSHI; WALIA; RANA, 2012; HOSSEINPOUR et al., 2012;PANESAR, KENNEDY, 2012;LIN et al., 2013;WANG et al., 2013;SUNG, 2013; AGGELOPOULOS et al., 2014;PLEISSNER et al., 2014;MARQUES et al., 2014 IDENTIFICAÇÃO E CARACTERIZAÇÃO MORFOLÓGICAA identificação e a caracterização morfológica da amostra foram realizadas através da metodologia descrita por Klick (2002), por meio da chave para identificação da espécie. A amostra foi cultivada nos meios Ágar Czapek autolisado de levedura -(CYE) (PITT, 1979) a 25 °C e 37 °C, e Extrato de Malte Àgar (MEA) a 25 °C. As características macroscópicas foram determinadas pela textura da colônia, grau de esporulação, cor dos conídios, textura e cor do micélio, presença de cores, pigmentos e exsudado.Para análise microscópica, foram preparadas lâminas, para observação das estruturas microscópicas como:comprimento, largura e textura dos conidióforos, forma da cabeça conidial, diâmetro da vesícula, comprimento das métulas e fiálides, diâmetro, forma, textura presença ou ausência de conídios e ascósporos, forma e cor do cleistotécio/esclerócios. PRÉ-INOCULOForam utilizadas placas de Petri contendo o meio ágar Sabouraud (acrescido de azeite de oliva para aclimatação) e incubadas a 28 ºC durante 7 dias. CINÉTICA DE CRESCIMENTO E DE PRODUÇÃO DE LIPASE EM MEIO CONVENCIONAL DETERMINAÇÃO DA BIOMASSA MICROBIANAA determinação da biomassa foi feita através de gravimetria. A massa micelial foi lavada e filtrada em papel de filtro, transferida para a estufa a 50 °C para secagem, em seguida transferida para o dessecador até peso constante. DETERMINAÇÃO DA ATIVIDADE LIPOLÍTICAA atividade lipolítica das amostras foi detectada utilizando-se a metodologia descrita por Soares et al. (1999). Através da reação de 5 mL de emulsão de óleo de oliva e goma arábica (7 %), 2 mL de tampão fosfato de sódio (0,1 M), pH 7,0 e 1 mL do líquido metabólico. A mistura foi agitada em agitador orbital a 82 rpm, 37 ºC, durante 10 minutos.A reação foi paralisada através da adição de 10 mL de EXTRAÇÃO E QUANTIFICAÇÃO DOS LIPÍDEOS TOTAISOs lipídeos totais da biomassa foram extraídos utilizando-se 1,0 g da biomassa liofilizada que foi submetida à extração com a mistur...

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Screening for novel lipolytic enzymes from uncultured soil microorganisms

Cited by 160 publications

References 39 publications

Cloning and Characterization of a Novel Carboxylesterase Gene from Cow Rumen Metagenomic Library

Cloning and Characterization of a Novel Carboxylesterase Gene from Cow Rumen Metagenomic Library

From bacterial genomics to metagenomics: concept, tools and recent advances

Avaliação Do Potencial Biotecnológico De Aspergillus Parasiticus Ucp 1281 No Biotratamento De Efluentes Da Indústria De Laticínios E Produção De Lipídeos

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