Dataset Information

Stability/activity tradeoffs in Thermusthermophilus HB27 laccase.

ABSTRACT: We report the temperature dependence of the formal potential of type 1 copper (Cu_T1) in Thermusthermophilus HB27 laccase. Employing [Ru(NH₃)₄(bpy)](PF₆)₂ (0.505 vs. NHE) as the redox titrant, we found that the Cu_T1^2+/+ potential decreased from approximately 480 to 420 mV (vs. NHE) as the temperature was raised from 20 to 65 °C. Of importance is that the ?S_rc° of?-?120 J mol^-1 K^-1 is substantially more negative than those for other blue copper proteins. We suggest that the highly unfavorable reduction entropy is attributable to Cu_T1 inaccessibility to the aqueous medium. Although the active site residues are buried, which is critical for maintaining thermostability, the flexibility around Cu_T1 is maintained, allowing enzyme activity at ambient temperature.

SUBMITTER: Shin J

PROVIDER: S-EPMC7085441 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Stability/activity tradeoffs in Thermusthermophilus HB27 laccase.

Shin Jieun J Gray Harry B HB Winkler Jay R JR

Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry 20200122 2

We report the temperature dependence of the formal potential of type 1 copper (CuT1) in Thermusthermophilus HB27 laccase. Employing [Ru(NH3)4(bpy)](PF6)2 (0.505 vs. NHE) as the redox titrant, we found that the CuT12+/+ potential decreased from approximately 480 to 420 mV (vs. NHE) as the temperature was raised from 20 to 65 °C. Of importance is that the ΔSrc° of - 120 J mol-1 K-1 is subs ...[more]

PMID: 31970489

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Project description:BackgroundLaccases (EC 1.10.3.2) are multi-copper oxidoreductases with great biotechnological importance due to their high oxidative potential and utility for removing synthetic dyes, oxidizing phenolic compounds, and degrading pesticides, among others.MethodsA real-time stability study (RTS) was conducted for a year, by using enzyme concentrates from 3 batches (L1, L3, and L4). For which, five temperatures 243.15, 277.15, 298.15, 303.15, 308.15, and 313.15 K were assayed. Using RTS data and the Arrhenius equation, we calculated the rPOXA 1B accelerated stability (AS). Molecular dynamics (MD) computational study results were very close to those obtained experimentally at four different temperatures 241, 278, 298, and 314 K.ResultsIn the RTS, 101.16, 115.81, 75.23, 46.09, 5.81, and 4.83% of the relative enzyme activity were recovered, at respective assayed temperatures. AS study, showed that rPOXA 1B is stable at 240.98 ± 5.38, 277.40 ± 1.32 or 297.53 ± 3.88 K; with t1/2 values of 230.8, 46.2, and 12.6 months, respectively. Kinetic and thermodynamic parameters supported the high stability of rPOXA 1B, with an Ed value of 41.40 KJ mol- 1, a low variation of KM and Vmax, at 240.98 ± 5.38, and 297.53 ± 3.88 K, and ∆G values showing deactivation reaction does not occur. The MD indicates that fluctuations in loop, coils or loops with hydrophilic or intermediate polarity amino acids as well as in some residues of POXA 1B 3D structure, increases with temperature; changing from three fluctuating residues at 278 K to six residues at 298 K, and nine residues at 314 K.ConclusionsLaccase rPOXA 1B demonstrated experimentally and computationally to be a stable enzyme, with t1/2 of 230.8, 46.2 or 12.6 months, if it is preserved impure without preservatives at temperatures of 240.98 ± 5.38, 277.40 ± 1.32 or 297.53 ± 3.88 K respectively; this study could be of great utility for large scale producers.

Dataset Information

Stability/activity tradeoffs in Thermusthermophilus HB27 laccase.

Publications

Stability/activity tradeoffs in Thermusthermophilus HB27 laccase.

Similar Datasets

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