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Harnessing hyperthermostable lactonase from Sulfolobus solfataricus for biotechnological applications

View Article: PubMed Central - PubMed

ABSTRACT

Extremozymes have gained considerable interest as they could meet industrial requirements. Among these, SsoPox is a hyperthermostable enzyme isolated from the archaeon Sulfolobus solfataricus. This enzyme is a lactonase catalyzing the hydrolysis of acyl-homoserine lactones; these molecules are involved in Gram-negative bacterial communication referred to as quorum sensing. SsoPox exhibits promiscuous phosphotriesterase activity for the degradation of organophosphorous chemicals including insecticides and chemical warfare agents. Owing to its bi-functional catalytic abilities as well as its intrinsic stability, SsoPox is appealing for many applications, having potential uses in the agriculture, defense, food and health industries. Here we investigate the biotechnological properties of the mutant SsoPox-W263I, a variant with increased lactonase and phosphotriesterase activities. We tested enzyme resistance against diverse process-like and operating conditions such as heat resistance, contact with organic solvents, sterilization, storage and immobilization. Bacterial secreted materials from both Gram-negative and positive bacteria were harmless on SsoPox-W263I activity and could reactivate heat-inactivated enzyme. SsoPox showed resistance to harsh conditions demonstrating that it is an extremely attractive enzyme for many applications. Finally, the potential of SsoPox-W263I to be active at subzero temperature is highlighted and discussed in regards to the common idea that hyperthermophile enzymes are nearly inactive at low temperatures.

No MeSH data available.


SsoPox-W263I relative activity after submission to three sterilization methods: autoclave (a), ethylene oxide (b), β-radiation (c). Experiments on solid enzyme are in brown bars while those on liquid enzyme are in blue. Measures were performed in triplicate as compared to a non-sterilized control. Values represent means ± SEM.
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f2: SsoPox-W263I relative activity after submission to three sterilization methods: autoclave (a), ethylene oxide (b), β-radiation (c). Experiments on solid enzyme are in brown bars while those on liquid enzyme are in blue. Measures were performed in triplicate as compared to a non-sterilized control. Values represent means ± SEM.

Mentions: Autoclaving is commonly used in medical applications for sterilizing materials. It uses a combination of elevated temperature and pressure and is usually destructive for proteins. Here we autoclaved the enzyme in liquid or in solid state (powder) at 121 °C for 15 minutes (Fig. 2a). Surprisingly, more than 30% activity remained after autoclaving the enzyme powder, while all the activity was lost in the case of the liquid form, which is consistent with the temperature resistance of the liquid enzyme.


Harnessing hyperthermostable lactonase from Sulfolobus solfataricus for biotechnological applications
SsoPox-W263I relative activity after submission to three sterilization methods: autoclave (a), ethylene oxide (b), β-radiation (c). Experiments on solid enzyme are in brown bars while those on liquid enzyme are in blue. Measures were performed in triplicate as compared to a non-sterilized control. Values represent means ± SEM.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5120315&req=5

f2: SsoPox-W263I relative activity after submission to three sterilization methods: autoclave (a), ethylene oxide (b), β-radiation (c). Experiments on solid enzyme are in brown bars while those on liquid enzyme are in blue. Measures were performed in triplicate as compared to a non-sterilized control. Values represent means ± SEM.
Mentions: Autoclaving is commonly used in medical applications for sterilizing materials. It uses a combination of elevated temperature and pressure and is usually destructive for proteins. Here we autoclaved the enzyme in liquid or in solid state (powder) at 121 °C for 15 minutes (Fig. 2a). Surprisingly, more than 30% activity remained after autoclaving the enzyme powder, while all the activity was lost in the case of the liquid form, which is consistent with the temperature resistance of the liquid enzyme.

View Article: PubMed Central - PubMed

ABSTRACT

Extremozymes have gained considerable interest as they could meet industrial requirements. Among these, SsoPox is a hyperthermostable enzyme isolated from the archaeon Sulfolobus solfataricus. This enzyme is a lactonase catalyzing the hydrolysis of acyl-homoserine lactones; these molecules are involved in Gram-negative bacterial communication referred to as quorum sensing. SsoPox exhibits promiscuous phosphotriesterase activity for the degradation of organophosphorous chemicals including insecticides and chemical warfare agents. Owing to its bi-functional catalytic abilities as well as its intrinsic stability, SsoPox is appealing for many applications, having potential uses in the agriculture, defense, food and health industries. Here we investigate the biotechnological properties of the mutant SsoPox-W263I, a variant with increased lactonase and phosphotriesterase activities. We tested enzyme resistance against diverse process-like and operating conditions such as heat resistance, contact with organic solvents, sterilization, storage and immobilization. Bacterial secreted materials from both Gram-negative and positive bacteria were harmless on SsoPox-W263I activity and could reactivate heat-inactivated enzyme. SsoPox showed resistance to harsh conditions demonstrating that it is an extremely attractive enzyme for many applications. Finally, the potential of SsoPox-W263I to be active at subzero temperature is highlighted and discussed in regards to the common idea that hyperthermophile enzymes are nearly inactive at low temperatures.

No MeSH data available.