Carboxylesterase from Sulfolobus solfataricus P1

Sehgal, Amitabh C.; Callen, Walter; Mathur, Eric J.; Short, Jay M.; Kelly, Robert M.

doi:10.1016/s0076-6879(01)30398-1

Cited by 19 publications

(22 citation statements)

References 32 publications

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“…The amino acid sequence predicted from the S. acidophilus esterase (EstS1) gene showed only 43%, 38%, and 26% identity with those from P. calidifontis, Sulfolobus solfataricus P1 (22), and Acinetobacter sp. M673 (also a PAE-degrading enzyme), respectively (Fig.…”

Section: Resultsmentioning

confidence: 99%

Newly Identified Thermostable Esterase from Sulfobacillus acidophilus: Properties and Performance in Phthalate Ester Degradation

Zhang

Fan

Qiu

et al. 2014

Appl Environ Microbiol

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EstS1, a newly identified thermostable esterase from Sulfobacillus acidophilus DSM10332, was heterologously expressed in Escherichia coli and shown to enzymatically degrade phthalate esters (PAEs) to their corresponding monoalkyl PAEs. The optimal pH and temperature of the esterase were found to be 8.0 and 70°C, respectively. The half-life of EstS1 at 60°C was 15 h, indicating that the enzyme had good thermostability. The specificity constant (k cat /K m ) of the enzyme for p-nitrophenyl butyrate was as high as 6,770 mM ؊1 s ؊1 . The potential value of EstS1 was demonstrated by its ability to effectively hydrolyze 35 to 82% of PAEs (10 mM) within 2 min at 37°C, with all substrates being completely degraded within 24 h. At 60°C, the time required for complete hydrolysis of most PAEs was reduced by half. To our knowledge, this enzyme is a new esterase identified from thermophiles that is able to degrade various PAEs at high temperatures.

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Section: Resultsmentioning

confidence: 99%

Newly Identified Thermostable Esterase from Sulfobacillus acidophilus: Properties and Performance in Phthalate Ester Degradation

Zhang

Fan

Qiu

et al. 2014

Appl Environ Microbiol

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“…The solution was refluxed for 12 h. The reaction mixture was extracted and purified by repeated extraction with methylene chloride and 5% sodium bicarbonate solution. The structure of synthesized methyl ester was confirmed by 13 C-NMR (500 MHz, DMSO-d 6 ) and 1H-NMR (500 MHz, DMSO-d 6 ). The racemic ketoprofen methyl ester was dissolved in methanol and added to 50 mM Tris-HCl, pH 7.0 (final substrate concentration 1 mM) containing the enzyme (0.234 mg).…”

Section: Methodsmentioning

confidence: 96%

“…11,12) The carboxylesterases from Sulfolobus solfataricus P1 and MT4 were also cloned and characterized. 13,14) But the lipolytic enzymes from S. solfataricus P2 have been not reported and their usability as biocatalysts is still unknown. In this paper, we report the characterization of the recombinant thermostable esterase (Est3) from S. solfataricus heterologously expressed in E. coli.…”

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confidence: 99%

Thermostable Esterase from a Thermoacidophilic Archaeon: Purification and Characterization for Enzymatic Resolution of a Chiral Compound

Kim

Lee

2004

Bioscience, Biotechnology, and Biochemistry

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“…Genomic DNA was prepared from S. solfataricus P1 (Sso P1) using a method similar to that described by (Cady et al, 2001). The gene encoding Sso EST1 was expressed using the vector pQE30 (Qiagen Inc., Valencia, CA) in Escherichia coli host strain, M15 pREP4 (Qiagen Inc., Valencia, CA) (Sehgal et al, 2001).…”

Section: Production Of Recombinant Sso Est1mentioning

confidence: 99%

“…The capability of increasing the functional temperature range of already stable biocatalysts would make these enzymes more attractive for many biotechnological purposes. This issue is explored here with the carboxylesterase from the extreme thermoacidophile Sulfolobus solfataricus P1 (Sso EST1) (Sehgal et al, 2001). …”

Section: Introductionmentioning

confidence: 99%

Structural and catalytic response to temperature and cosolvents of carboxylesterase EST1 from the extremely thermoacidophilic archaeon Sulfolobus solfataricus P1

Sehgal

Tompson

Cavanagh

et al. 2002

Biotech & Bioengineering

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The interactive effects of temperature and cosolvents on the kinetic and structural features of a carboxylesterase from the extremely thermoacidophilic archaeon Sulfolobus solfataricus P1 (Sso EST1) were examined. While dimethylformamide, acetonitrile, and dioxane were all found to be deleterious to enzyme function, dimethyl sulfoxide (DMSO) activated Sso EST1 to various extents. This was particularly true at 3.5% (v/v) DMSO, where k(cat) was 20-30% higher than at 1.2% DMSO, over the temperature range of 50-85 degrees C. DMSO compensated for thermal activation in some cases; for example, k(cat) at 60 degrees C in 3.5% DMSO was comparable to k(cat) at 85 degrees C in 1.2% DMSO. The relationship between DMSO activation and enzyme structural characteristics was also investigated. Nuclear magnetic resonance spectroscopy and circular dichroism showed no gross change in enzyme conformation with 3.5% DMSO between 50 and 80 degrees C. However, low levels of DMSO were shown to have a small yet significant change in enzyme conformation. This was evident through the reduction of Sso EST1's melting temperature and changes in the microenvironment of the enzyme's tyrosine and tryptophan residues at 3.5% versus 1.2% (v/v) solvent. Finally, activation parameter analysis based on kinetic data, at 1.2% and 3.5% DMSO, implied an increase in conformational flexibility with additional cosolvent. These results suggest the activating effect of DMSO was related to small changes in the enzyme's structure resulting in an increase in its conformational flexibility. Thus, in addition to their use for solubilizing hydrophobic substrates in water, cosolvents may also serve as activators in applications involving thermostable biocatalysts at sub-optimal temperatures.

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Carboxylesterase from Sulfolobus solfataricus P1

Cited by 19 publications

References 32 publications

Newly Identified Thermostable Esterase from Sulfobacillus acidophilus: Properties and Performance in Phthalate Ester Degradation

Newly Identified Thermostable Esterase from Sulfobacillus acidophilus: Properties and Performance in Phthalate Ester Degradation

Thermostable Esterase from a Thermoacidophilic Archaeon: Purification and Characterization for Enzymatic Resolution of a Chiral Compound

Structural and catalytic response to temperature and cosolvents of carboxylesterase EST1 from the extremely thermoacidophilic archaeon Sulfolobus solfataricus P1

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