Characterization of a novel oligomeric SGNH-arylesterase from Sinorhizobium meliloti 1021

Hwang, Heejin; Kim, Seungbum; Yoon, Sangyoung; Ryu, Yeon‐Woo; Lee, Sang Yoon; Kim, T. Doohun

doi:10.1016/j.ijbiomac.2009.12.010

Cited by 24 publications

(16 citation statements)

References 35 publications

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“…Accordingly, we mutated this residue to Ala ad check the hydrolysis activity of the mutant for confirming this hypothesis. Oxyanion hole components (Ser, Gly/Ala, Asn, and His) in the known CE3 (Est24, Sm23, CtCES3_1, and TcAE206) are also conserved in CtCE2 and SmAcE1 (Figure 1) [27][28][29][30][31]. From these analyses, it can be proposed that SmAcE1 possibly has a similar catalytic mechanism to the known carbohydrate esterases belonging to CE2 and CE3.…”

Section: Resultsmentioning

confidence: 86%

“…Sm23 (PDB code: 4TX1, [25]), Est24 (PDB code: 5HOE [26]), TcAE206 (PDB code: 5B5S [27]), and CtCES3-1 (PDB code: 2TVP [28]) were used. Furthermore, CtCE2 from Clostridium thermocellum (PDB code: 2WAO [29]) that belongs to the CE2 was also included for the sequence alignment [30,31]. Amino acid sequences were obtained from UniProt (http://www.uniprot.org/) and sequence alignment was done using PROMALS3D [32] and were visualized using ESPript [33].…”

Section: Sequence Alignment Of Smace1 With Other Cesmentioning

confidence: 99%

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Identification and Crystallographic Analysis of a New Carbohydrate Acetylesterase (SmAcE1) from Sinorhizobium meliloti

Ryu

Yoo

et al. 2018

Crystals

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Carbohydrate-active enzymes (CAZymes) regulate the synthesis, degradation, and modification of the poly-and oligosaccharides in all three kingdoms of life. A novel carbohydrate acetylesterase from Sinorhizobium meliloti, designated SmAcE1, was identified, characterized, and crystallized. This SmAcE1 is classified into the carbohydrate esterase family 3 (CE3) based on the sequence alignments with other currently known carbohydrate esterase (CE) family enzymes. The SmAcE1 was crystallized as a hexamer in a space group P2 1 2 1 2 1 with the unit cell parameters: a = 99.12 Å, b = 148.88 Å, c = 149.84 Å, and α = β = γ = 90.00 • . The diffraction data set was collected up to a 2.05 Å resolution. Hydrolysis activity of SmAcE1 towards glucose pentaacetate and cellulose acetate was further confirmed using acetic acid release assay. Further crystallographic and functional analyses studies on SmAcE1 would be followed to fully understand the reaction mechanisms of CEs.

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Section: Resultsmentioning

confidence: 86%

Section: Sequence Alignment Of Smace1 With Other Cesmentioning

confidence: 99%

Identification and Crystallographic Analysis of a New Carbohydrate Acetylesterase (SmAcE1) from Sinorhizobium meliloti

Ryu

Yoo

et al. 2018

Crystals

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“…presenta la máxima actividad a pH 8. Este dato concuerda con el de otras esterasas como la EstGtA2 (Charbonneau et al 2010), la Sm23 (Hwang et al 2010) o la esterasa de Sulfolobus tokodaii (Suzuki et al 2004). …”

Section: Características Cinéticasunclassified

“…Comparando los resultados obtenidos con los de esterasas termoestables como son la Est3 (Kim & Lee. 2004) y la Sm23 (Hwang et al 2010), la Est2528 es más estable en presen-84 I. Navarro-González Anales de Biología 34, 2012 cia de detergentes que la Sm23, mientras que en el caso de la Est3 es más estable que la Est2528 solo en presencia de Triton X-100, lo que indica que la Est2528 es una buena candidata para su posible aplicación industrial. Del estudio llevado a cabo con los disolventes orgánicos se observa que con todos los usados (etanol, propanol, metanol, DMSO, acetona y acetonitrilo) al 5 % (V/V) la enzima es estable.…”

Section: Efecto De Los Detergentes Y De Los Disolventes Orgánicos Sobunclassified

“…2004) y la Sm23 (Hwang et al 2010), se pone de manifiesto que la Est2528 es mucho más estable, lo que puede hacerla más atractiva industrialmente, ya que a la temperatura a la cual se utilizan las esterasas en algunos procesos tecnológicos las proteínas mesófilas tiene una mayor actividad que las proteínas termófilas. La ventaja que presentan la mayoría de las proteínas termoestables frente a las mesófilas, es su resistencia a desnaturalizarse en condiciones extremas como es el exponerlas a elevadas concentraciones de disolventes orgánicos.…”

Section: Efecto De Los Detergentes Y De Los Disolventes Orgánicos Sobunclassified

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Clonacion, expresión, caracterización y modelado de esterasas putativas de Corynebacterium glutamicum ATTC 13032

Inmaculada¹

2012

analesbio

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El objetivo de este trabajo fue clonar, expresar, purificar y caracterizar algunos genes de Corynebacterium glutamicum ATCC13032, identificados como enzimas de la superfamilia de las esterasas/lipasas. De todos los genes clonados, solo se pudo estudiar la proteína codificada por el gen Cg2528. Se purificó y caracterizó la proteína. Su máxima actividad la presentó 40 ºC y a pH 8. Presentó una gran estabilidad en presencia de disolventes orgánicos como el DMSO y el metanol. Con respecto al análisis cinético, presentó una mayor actividad sobre los p-nitrofenil ésteres y los naftilos que sobre el S-methiltiobutanoato. Se dilucidó la triada catalíti-ca alineando la secuencia de aminoácidos. La estructura tridimensional se predijo usando la esterasa (2OGS) de Geobacillus stearothermophilus como molde.Palabras clave: Esterasas, Purificación, Caracterización bioquímica, Clonación. Abstract Cloning, expression, characterization and modeling of putatives esterases from Corynebacterium glutamicum ATCC 13032The aim of this work was to clone, express, purifiy and characterize some genes from Corynebacterium glutamicum ATCC13032, identified as enzymes of the superfamily of esterases/lipases. Among all cloned genes, only the protein encoded by the gene Cg-2528 could be studied. The protein was purified and characterized. The optimum activity was observed at 40ºC and pH 8. The enzyme showed a high activity in presence of organic solvents, such as DMSO or methanol. P-nitrophenyl ester and naphthyl ester derivatives were found to be better substrates than S-methyl tiobutanoate. The catalytic triad was predicted by amino acid sequence alignment and the structure modeling was performed using esterase (2OGS) form Geobacillus stearothermophilus as template.

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Molecular cloning and characterization of a novel SGNH arylesterase from the goat rumen contents

Wang

Meng

Luo

et al. 2011

Appl Microbiol Biotechnol

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An esterase-encoding gene, estR5, was directly obtained from the genomic DNA of goat rumen contents. The 555-bp full-length gene encodes a 184-residue polypeptide (EstR5) without putative signal peptide. Deduced EstR5 shared the highest identity (50%) to a putative arylesterase from Ruminococcaceae bacterium D16. Phylogenetic analysis indicated that EstR5 was closely related with microbial esterases of gastrointestinal source. A comparison of the conserved motifs shared with GDSL proteins revealed that EstR5 could be grouped into the GDSL family and was further classified into the subfamily of SGNH hydrolases. The gene estR5 was expressed in Escherichia coli BL21 (DE3) and purified to electrophoretic homogeneity. Recombinant EstR5 exhibited highest catalytic efficiency towards α-naphthyl acetate followed by phenyl acetate and p-nitrophenyl acetate and had no activity towards PNP esters with acyl chains longer than C8. The enzyme exhibited optimal activity at around 60°C and pH 8.0, was stable at pH ranging from 6.0 to 11.0 and was slightly activated by detergent Tween, Nonidet P-40, and Triton X-100. These properties suggest that EstR5 has great potential for basic research and industrial applications. To our knowledge, this is the first arylesterase obtained from rumen microenvironment.

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Characterization of a novel oligomeric SGNH-arylesterase from Sinorhizobium meliloti 1021

Cited by 24 publications

References 35 publications

Identification and Crystallographic Analysis of a New Carbohydrate Acetylesterase (SmAcE1) from Sinorhizobium meliloti

Identification and Crystallographic Analysis of a New Carbohydrate Acetylesterase (SmAcE1) from Sinorhizobium meliloti

Clonacion, expresión, caracterización y modelado de esterasas putativas de Corynebacterium glutamicum ATTC 13032

Molecular cloning and characterization of a novel SGNH arylesterase from the goat rumen contents

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