Use of Schizosaccharomyces strains for wine fermentation? Effect on the wine composition and food safety

Schizosaccharomyceswas initially considered as a spoilage yeast because of the production of undesirable metabolites such as acetic acid, hydrogen sulfide, or acetaldehyde, but it currently seems to be of great value in enology.o ced Nevertheless, Schizosaccharomyces can reduce all of the malic acid in must, leading to malolactic fermentation. Malolactic fermentation is a highly complicated process in enology and leads to a higher concentration of biogenic amines, so the use of Schizosaccharomyces pombe can be an excellent tool for assuring wine safety. Schizosaccharomyces also has much more potential than only reducing the malic acid content, such as increasing the level of pyruvic acid and thus the vinylphenolic pyranoanthocyanin content. Until now, few commercial strains have been available and little research on the selection of appropriate yeast strains with such potential has been conducted. In this study, selected and wild Sc. pombe strains were used along with a Saccharomyces cerevisiae strain to ferment red grape must. The results showed significant differences in several parameters including non-volatile and volatile compounds, anthocyanins, biogenic amines and sensory parameters

Selected Schizosaccharomyces pombe Strains Have Characteristics That Are Beneficial for Winemaking

At present, wine is generally produced using Saccharomyces yeast followed by Oenococus bacteria to complete malolactic fermentation. This method has some unsolved problems, such as the management of highly acidic musts and the production of potentially toxic products including biogenic amines and ethyl carbamate. Here we explore the potential of the fission yeast Schizosaccharomyces pombe to solve these problems. We characterise an extensive worldwide collection of S. pombe strains according to classic biochemical parameters of oenological interest. We identify three genetically different S. pombe strains that appear suitable for winemaking. These strains compare favourably to standard Saccharomyces cerevisiae winemaking strains, in that they perform effective malic acid deacidification and significantly reduce levels of biogenic amines and ethyl carbamate precursors without the need for any secondary bacterial malolactic fermentation. These findings indicate that the use of certain S. pombe strains could be advantageous for winemaking in regions where malic acid is problematic, and these strains also show superior performance with respect to food safety

Biotechnological applications of indigenous wine yeasts based on genomic and proteonic analysis

Several indigenous yeast strains were isolated from spontaneously fermenting Debina must. All isolates were taxonomically characterized using molecular approaches i.e. sequencing of the variable ITS1 - 5.8S rDNA - ITS2 and D1/D2 regions. Based on these analyses, the isolates from Sample I, exhibiting β-glucosidase activity, were identified as: H. gulliermondii (ΖΥ1, ΖΥ2, ΖΥ3), R. mucilaginosa (ΖΥ5), P. anomala (ΖΥ4) and M. pulcherrima, var. zitsae (ΖΥ6). Regarding the isolates of Sample II, all indigenous strains proved to belong to the species S. cerevisiae. Proceeding with the investigation of the impact of the indigenous yeast on wine characteristics, various isolated strains were used as starter cultures for the production of the local Debina wine. Strain Z622 (S. cerevisiae) served as a starter in year 2005, while in years 2006 and 2007 non-Saccharomyces strains (ZY6 or a mixture of ZY3, ZY4) and Z622 (S. cerevisiae) were used in a sequential fashion. In every purpose, a second tank containing the same must inoculated with a S. cerevisiae commercial starter culture, routinely used by the winery, served as a control vinification. A comparative study of the produced wines concerning their physicochemical features, as well as their profiles of several volatile compounds, revealed that the wine produced with the indigenous strains exhibited the best organoleptic characteristics and the richest aromatic bouquet. With regard to the evaluation of the proteome changes and the identification of those proteins induced in Debina-must fermentation we compared the 2D PAGE maps of strain Z622 cells grown in three different media (YM, MM, Debina must). The 14 intracellular yeast proteins induced only during must fermentation were identified unambiguously by MS analysis, apparently involved in various metabolic pathways such as glycolysis, amino-acid metabolism or production of antioxidants. was selected because as compared to the rest In order to verify the proteomics data, we studied the induction of the bi-functional O-acetylhomoserine sulfhydrylase/ O-acetylserine sulfhydrylase on the transcriptional level employing a reverse transcriptional-PCR analysis. This enzyme is considered the most important for the wine industry due to its involvement in the limitation of reduced sulfides in the wine. This study underlines the variation in pathway regulation under fermentation conditions, implying the importance of a thorough understanding of this complex procedure.Κατά τη διάρκεια αυθόρμητης οινοποίησης γλεύκους Ντεμπίνα απομονώθηκαν στελέχη ζυμομυκήτων από τη Φάση Ι: αρχή της ζύμωσης και από τη Φάση ΙΙ: τέλος της ζύμωσης. Τα στελέχη που απομονώθηκαν ταυτοποιήθηκαν με βάση τους πολυμορφισμούς της περιοχών ITS1-5.8S rDNA -ITS2 και D1/D2. Από τη Φάση Ι ταυτοποιήθηκαν έξι στελέχη τα οποία εμφάνισαν δραστικότητα β-γλυκοζιδάσης και τα οποία ταξινομήθηκαν ως H. gulliermondii (ΖΥ1, ΖΥ2, ΖΥ3), R. mucilaginosa (ΖΥ5), P. anomala (ΖΥ4) και M. pulcherrima, var. zitsae (ΖΥ6). Από τη Φάση ΙΙ απομονώθηκαν πέντε στελέχη Ζ607, Ζ608, Ζ614, Ζ617 και Ζ622 τα οποία ταξινομήθηκαν ως στελέχη του είδους S. cerevisiae. Με σκοπό τη μελέτη της επίδρασης των γηγενών στελεχών ζυμομυκήτων στον αρωματικό χαρακτήρα οίνων Ντεμπίνα, χρησιμοποιήσαμε τα απομονωθέντα στελέχη ως καλλιέργειες εκκίνησης. Το έτος 2005 χρησιμοποιήθηκε το στέλεχος Ζ622, ενώ τα έτη 2006 και 2007 χρησιμοποιήθηκε μεθοδολογία διαφορικής οινοποίησης με τα στελέχη non-Saccharomyces (ΖΥ6 ή μίγμα ΖΥ3 και ΖΥ4) να λειτουργούν ως καλλιέργειες εκκίνησης και το στέλεχος Ζ622 (S.cerevisiae) να χρησιμεύει ως καλλιέργεια ολοκλήρωσης. Ως μάρτυρας χρησιμοποιήθηκε οινοποίηση του ιδίου γλεύκους με ένα εμπορικό στέλεχος του είδους S.cerevisiae. Η οργανοληπτική και αεριοχρωματογραφική ανάλυση των παραχθέντων οίνων αποκάλυψε ότι οι παραχθέντες οίνοι με τα γηγενή οινοποιητικά στελέχη είχαν καλύτερα οργανοληπτικά χαρακτηριστικά και πλουσιότερο αρωματικό δυναμικό από τους αντίστοιχους οίνους μάρτυρες, επιβεβαιώνοντας την άποψη ότι τα γηγενή στελέχη ζυμών κάθε αμπελώνα είναι αυτά που προδίδουν στον οίνο τα ιδιαίτερα χαρακτηριστικά του. Στοχεύοντας στη διερεύνηση των βιοχημικών πορειών που επάγονται κατά τη ζύμωση γλεύκους, πραγματοποιήθηκε με τη βοήθεια της Ηλεκτροφόρησης δύο διαστάσεων, συγκριτική πρωτεωμική ανάλυση των κυττάρων του στελέχους Ζ622 που ζύμωσαν τρία διαφορετικά θρεπτικά υποστρώματα YΜ, ΜΜ και Γλεύκος. Τα επαγόμενα πρωτεϊνικά μόρια μόνο κατά τη ζύμωση γλεύκους που ταυτοποιήθηκαν με μεθοδολογία LC-MS/MS εμπλέκονται σε διάφορες μεταβολικές πορείες όπως η γλυκόλυση, ο μεταβολισμός αμινοξέων, η παραγωγή αντιοξειδωτικών κ.ά Από τα ανιχνευθέντα ένζυμα το κυριότερο για τις οινοποιητικές βιομηχανίες είναι η σουλφυδρυλάση της Ο-ακετυλο-σερίνης/Ο-ακετυλο-ομοσερίνης, λόγω του ότι η επαγωγή του ή όχι συνδέεται με τη συγκέντρωση του H2S, μιας ουσίας της οποίας η δυσάρεστη οσμή μπορεί να καταστρέψει τον οίνο. Τα αποτελέσματα της πρωτεωμικής ανάλυσης επιβεβαιώθηκαν και στο μεταγραφικό επίπεδο με τη χρήση μεθοδολογίας RT-RCR

Correction to: Molecular, biochemical and kinetic analysis of a novel, thermostable lipase (LipSm) from Stenotrophomonas maltophilia Psi‑1, the first member of a new bacterial lipase family (XIX)

We have recently (8th February 2018) published our article entitled “Molecular, biochemical and kinetic analysis of a novel, thermostable lipase (LipSm) from Stenotrophomonas maltophilia Psi-1, the first member of a new bacterial lipase family (XVIII)” [1]. While our manuscript was going through the final stages of publication, an article by Samoylova et al. [2] was published (12th January 2018) in the journal Extremophiles, entitled “Cloning, expression and characterization of the esterase estUT1 from Ureibacillus thermosphaericus which belongs to a new lipase family XVIII”. Since we could not have known of the work of Samoylova et al. [2] when we submitted our manuscript, and in order to avoid confusion in the scientific community, we propose to reclassify LipSm as the first characterized member of the new bacterial lipase family XIX. Therefore throughout our article [1] “lipase family XVIII” should read “lipase family XIX” (title included)

Kinetic constraints and features imposed by the immobilization of enzymes onto solid matrices: A key to advanced biotransformation

1045-1051The kinetics of immobilized enzymes can not be analyzed by means of the simple Michaelis-Menten concept, which generally fails to describe the immobilized state due to both its probable barriers, and because the active concentration of the enzyme approaches, or even exceeds this of its substrate(s). In such cases, the various experimental data are usually treated by complex rate equations comprising too many parameters acquiring different natures and meanings, depending on both the properties of the immobilization state and the experimental conditions; thus, more likely, only apparent values of the Michaelis-Menten kinetic parameters can be estimated experimentally. Likewise, immobilization is often a key method in optimizing the operational performance of enzymes, in both laboratory and industrial scale, and affects considerably the kinetics in non-aqueous and non-conventional media due to several issues as the structural changes of the enzyme molecule, the heterogeneity of the system, and the partial or total absence of water. In this work a theoretical approach is described on the formulation of simplified rate equations, reflecting also the actual mass balances of the reactants, in the case where esterification synthetic reactions are catalyzed by immobilized lipases, in either a non-aqueous organic solvent or in a non-solvent system

MOESM2 of Molecular, biochemical and kinetic analysis of a novel, thermostable lipase (LipSm) from Stenotrophomonas maltophilia Psi-1, the first member of a new bacterial lipase family (XVIII)

Additional file 2. Effect of metallic salts on the Michaelis–Menten parameters of LipSm