Characterization of a new thermophilic and acid tolerant esterase from Thermotoga maritima capable of hydrolytic resolution of racemic ketoprofen ethyl ester

Wei, Tao; Feng, Shengxue; Mao, Duobin; Xuan, Yue; Du, Congcong; Wang, Xihua

doi:10.1016/j.molcatb.2012.08.006

Cited by 21 publications

(6 citation statements)

References 46 publications

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“…Particularly, block H comprises two families: plant carboxylesterases and HSL, and its members are ubiquitous (see ESTHER database) (De Simone et al., ; Lenfant et al., ). According with this, it is not rare that PDB 2O7R was identified by I‐TASSER as a template during the first Ba EstB modeling round, probably being Ba EstB a member of block H. The HSL family is composed of esterases and lipases, and its members are widely represented in bacteria, plants, and animals (Tao et al., ). The HSL sequences identified in bacteria share high amino acid sequence similarity with the LIPE genes, which encode mammalian HSLs (Holm, ; Holm, Osterlund, Laurell, & Contreras, ).…”

Section: Resultsmentioning

confidence: 99%

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The first description of a hormone‐sensitive lipase from a basidiomycete: Structural insights and biochemical characterization revealed Bjerkandera adusta BaEstB as a novel esterase

et al. 2017

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The heterologous expression and characterization of a Hormone‐Sensitive Lipases (HSL) esterase (BaEstB) from the Basidiomycete fungus Bjerkandera adusta is reported for the first time. According to structural analysis, amino acid similarities and conservation of particular motifs, it was established that this enzyme belongs to the (HSL) family. The cDNA sequence consisted of 969 nucleotides, while the gene comprised 1133, including three introns of 57, 50, and 57 nucleotides. Through three‐dimensional modeling and phylogenetic analysis, we conclude that BaEstB is an ortholog of the previously described RmEstB‐HSL from the phylogenetically distant fungus Rhizomucor miehei. The purified BaEstB was characterized in terms of its specificity for the hydrolysis of different acyl substrates confirming its low lipolytic activity and a noticeable esterase activity. The biochemical characterization of BaEstB, the DLS analysis and the kinetic parameters determination revealed this enzyme as a true esterase, preferentially found in a dimeric state, displaying activity under alkaline conditions and relative low temperature (pH = 10, 20°C). Our data suggest that BaEstB is more active on substrates with short acyl chains and bulky aromatic moieties. Phylogenetic data allow us to suggest that a number of fungal hypothetical proteins could belong to the HSL family.

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Section: Resultsmentioning

confidence: 99%

“…The HSL family is composed of esterases and lipases, and its members are widely represented in bacteria, plants, and animals (Tao et al, 2013). The HSL sequences identified in bacteria share high amino acid sequence similarity with the LIPE genes, which encode mammalian HSLs (Holm, 2003;Holm, Osterlund, Laurell, & Contreras, 2000).…”

Section: Baestb Modelingmentioning

confidence: 99%

The first description of a hormone‐sensitive lipase from a basidiomycete: Structural insights and biochemical characterization revealed Bjerkandera adusta BaEstB as a novel esterase

et al. 2017

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“…Thus, TM1022 has a better pH stability. After incubation at 90 °C for 8 h, TM1022 retained 57% of its enzymatic activity, showing higher thermal stability than other thermophilic carboxylesterases. − The potent inhibition of PMSF suggests that serine residues are potentially associated with the catalytic site of TM1022, consistent with the predicted catalytic triad (Ser-Asp-His). Experiments on the effects of surfactants and organic solvents showed that TM1022 is very stable when exposed to chemical reagents.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Thermotoga maritima Esterase Capable of Hydrolyzing Bis(2-hydroxyethyl) Terephthalate

Feng,

Xue,

Xie

et al. 2024

J. Agric. Food Chem.

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The gene-encoding carboxylesterase (TM1022) from the hyperthermophilic bacterium Thermotoga maritima (T. maritima) was cloned and expressed in Escherichia coli Top10 and BL21 (DE3). Recombinant TM1022 showed the best activity at pH 8.0 and 85 °C and retained 57% activity after 8 h cultivation at 90 °C. TM1022 exhibited good stability at pH 6.0−9.0, maintaining 53% activity after incubation at pH 10.0 and 37 °C for 6 h. The esterase TM1022 exhibited the optimum thermo-alkali stability and k cat /K m (598.57 ± 19.97 s −1 mM −1 ) for pN−C4. TM1022 hydrolyzed poly(ethylene terephthalate) (PET) degradation intermediates, such as bis(2-hydroxyethyl) terephthalate (BHET) and mono(2-hydroxyethyl) terephthalate (MHET). The K m , k cat , and k cat /K m values for BHET were 0.82 ± 0.01 mM, 2.20 ± 0.02 s −1 , and 2.67 ± 0.02 mM −1 s −1 , respectively; those for MHET were 2.43 ± 0.07 mM, 0.04 ± 0.001 s −1 , and 0.02 ± 0.001 mM −1 s −1 , respectively. When purified TM1022 was added to the cutinase BhrPETase, hydrolysis of PET from drinking water bottle tops produced pure terephthalic acids (TPA) with 166% higher yield than those obtained after 72 h of incubation with BhrPETase alone as control. The above findings demonstrate that the esterase TM1022 from T. maritima has substantial potential for depolymerizing PET into monomers for reuse.

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“…The need for stable enzymes that tolerate alkaline conditions also justifies the screening and characterization of extremozymes (Box 1). Thermostable lipases of archaeal origin that prefer slightly alkaline conditions were recently identified from Sulfolobus acidophilus and Pyrobaculum sp., while lipolytic thermozymes that are optimally active at acidic conditions were exclusively isolated from Thermotoga maritima and Thermus thermophilus (Table 1) [35][36][37][38]. Especially the esterases from S. acidophilus and Pyrobaculum sp.…”

Section: Lipolytic Enzymesmentioning

confidence: 99%

Exploration of extremophiles for high temperature biotechnological processes

Elleuche

Schäfers

Blank

et al. 2015

Current Opinion in Microbiology

119

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Characterization of a new thermophilic and acid tolerant esterase from Thermotoga maritima capable of hydrolytic resolution of racemic ketoprofen ethyl ester

Cited by 21 publications

References 46 publications

The first description of a hormone‐sensitive lipase from a basidiomycete: Structural insights and biochemical characterization revealed Bjerkandera adusta BaEstB as a novel esterase

The first description of a hormone‐sensitive lipase from a basidiomycete: Structural insights and biochemical characterization revealed Bjerkandera adusta BaEstB as a novel esterase

Characterization of Thermotoga maritima Esterase Capable of Hydrolyzing Bis(2-hydroxyethyl) Terephthalate

Exploration of extremophiles for high temperature biotechnological processes

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