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Abstract
Anoxybacillus amylolyticus is a moderate thermophilic microorganism producing an exopolysaccharide and an extracellular α-amylase able to hydrolyze starch. The synthesis of several biomolecules is often regulated by a quorum sensing (QS) mechanism, a chemical cell-to-cell communication based on the production and diffusion of small molecules named “autoinducers”, most of which belonging to the N-acyl homoserine lactones’ (AHLs) family. There are few reports about this mechanism in extremophiles, in particular thermophiles. Here, we report the identification of a signal molecule, the N-butanoyl-homoserine lactone (C4-HSL), from the milieu of A. amylolyticus. Moreover, investigations performed by supplementing a known QS inhibitor, trans-cinnamaldehyde, or exogenous C4-HSL in the growth medium of A. amylolyticus suggested the involvement of QS signaling in the modulation of extracellular α-amylase activity. The data showed that the presence of the QS inhibitor trans-cinnamaldehyde in the medium decreased amylolytic activity, which, conversely, was increased by the effect of exogenous C4-HSL. Overall, these results represent the first evidence of the production of AHLs in thermophilic microorganisms, which could be responsible for a communication system regulating thermostable α-amylase activity.
Highlights
Accepted: 10 April 2021Anoxybacillus amylolyticus is a moderate thermophilicGram-positive rod and facultative anaerobe
The ethyl acetate extract (100 μL of a solution 0.6 mg mL−1 in methanol) of spent medium of A. amylolyticus grown in standard conditions was analyzed by means of thin-layer chromatography (TLC) overlay assay, with the biosensor Agrobacterium tumefaciens NTL4
Since A. amylolyticus is able to produce an extracellular amylase enzyme, we investigated the involvement of the quorum sensing (QS) mechanism in the extracellular amylolytic activity, comparing the C4-HSL level in a control growth, in presence of a QS inhibitor, trans-cinnamaldehyde (CIN) [14], or of exogenous C4-HSL
Summary
Accepted: 10 April 2021Anoxybacillus amylolyticus (strain MR3CT =ATCC BAA-872T ) is a moderate thermophilicGram-positive rod and facultative anaerobe. It is able to produce an exopolysaccharide and to synthesize an extracellular constitutive amylolytic activity which digests starch. The maximum amylolytic activity is expressed in the stationary growth phase [1]. Amylases belong to families of enzymes used in several biotechnological and industrial processes (e.g., starch processing, sugar production) [2,3], and in textile and pharmaceutical industries [4]. They can be obtained from different sources, including plants, animals, and microorganisms, microbial enzymes are generally preferred, due to their advantages such as efficient production, stability, and cost-effectiveness [5,6]
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