Proteomic analysis and optimized production of Alkalihalobacillus patagoniensis PAT 05 T extracellular proteases
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RESEARCH PAPER
Proteomic analysis and optimized production of Alkalihalobacillus patagoniensis PAT 0 5T extracellular proteases Nelda L. Olivera1 · Cynthia Sequeiros2 · Martín Iglesias1 · Marina Nievas2 Received: 20 May 2020 / Accepted: 25 August 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Extracellular proteolytic extracts from the haloalkalitolerant strain Alkalihalobacillus patagoniensis PAT 05T have proved highly efficient to reduce wool felting, as part of an ecofriendly treatment suitable for organic wool. In the present study, we identified the extracellular proteases produced by PAT 05T and we optimized its growth conditions for protease production through statistical methods. A total of 191 proteins were identified in PAT 0 5T culture supernatants through mass spectrometry analysis. Three of the 6 detected extracellular proteases belonged to the serine-endopeptidase family S8 (EC 3.4.21); two of them showed 86.3 and 67.9% identity with an alkaline protease from Bacillus alcalophilus and another one showed 50.4% identity with Bacillopeptidase F. The other 3 proteases exhibited 55.3, 49.4 and 61.1% identity with D-alanyl-Dalanine carboxypeptidase DacF, D-alanyl-D-alanine carboxypeptidase DacC and endopeptidase LytE, respectively. Using a Fractional Factorial Design followed by a Central Composite Design optimization, a twofold increase in protease production was reached. NaCl concentration was the most influential factor on protease production. The usefulness of PAT 0 5T extracellular proteolytic extracts to reduce wool felting was possible associated with the activity of the serine-endopeptidases closely related to highly alkaline keratinolytic proteases. The other identified proteases could cooperate, improving protein hydrolysis. This study provided valuable information for the exploitation of PAT 0 5T proteases which have potential for the valorization of organic wool as well as for other industrial applications. Graphic abstract
Keywords Extracellular proteome · LC‑MS/MS · Proteolytic activity · Response surface methodology · Wool felting
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00449-020-02436-z) contains supplementary material, which is available to authorized users. * Nelda L. Olivera olivera@cenpat‑conicet.gob.ar Extended author information available on the last page of the article
Introduction Alkalihalobacillus patagoniensis PAT 0 5T (basionym Bacillus patagoniensis) was isolated during a microorganism screening focused on novel strains from alkaline and saline Patagonian soils [1, 2]. In the early studies, PAT 05T showed
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extracellular proteolytic activity under high alkaline and saline conditions, which along with its residual activity at moderate temperature, suggested the potential of this strain as source of alkaline proteases [3]. More recently, PAT 0 5T extracellular proteases showed to be useful for wool antifelting treatment [4]. Wool is a natural fibre mainl
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