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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.


Immobilization rate of enzyme by crosslinking in alginate beads as compared to the free enzyme and recovery.The results for each cycle are represented with mean ± SEM in black bars. Immobilized and released activities are shown in brown and grey respectively.
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f7: Immobilization rate of enzyme by crosslinking in alginate beads as compared to the free enzyme and recovery.The results for each cycle are represented with mean ± SEM in black bars. Immobilized and released activities are shown in brown and grey respectively.

Mentions: We investigated the immobilization of SsoPox-W263I through crosslinking aggregation into alginate beads. Glutaraldehyde (GAD) was previously reported as an aggregating agent53, and was used here for crosslinking the enzyme inside alginate beads. Only a small amount of enzyme leaked out of the beads during the rinsing steps and the enzyme remained active and accessible after entrapment. Up to 15% of activity was immobilized with this technique. Bead recovery was subsequently investigated and the immobilized enzyme was used ten times with only 30% decrease (Fig. 7). This method offers a simple procedure for the cheap and efficient immobilization of SsoPox-W263I.


Harnessing hyperthermostable lactonase from Sulfolobus solfataricus for biotechnological applications
Immobilization rate of enzyme by crosslinking in alginate beads as compared to the free enzyme and recovery.The results for each cycle are represented with mean ± SEM in black bars. Immobilized and released activities are shown in brown and grey respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f7: Immobilization rate of enzyme by crosslinking in alginate beads as compared to the free enzyme and recovery.The results for each cycle are represented with mean ± SEM in black bars. Immobilized and released activities are shown in brown and grey respectively.
Mentions: We investigated the immobilization of SsoPox-W263I through crosslinking aggregation into alginate beads. Glutaraldehyde (GAD) was previously reported as an aggregating agent53, and was used here for crosslinking the enzyme inside alginate beads. Only a small amount of enzyme leaked out of the beads during the rinsing steps and the enzyme remained active and accessible after entrapment. Up to 15% of activity was immobilized with this technique. Bead recovery was subsequently investigated and the immobilized enzyme was used ten times with only 30% decrease (Fig. 7). This method offers a simple procedure for the cheap and efficient immobilization of SsoPox-W263I.

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.