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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Microbiology".

Deadline for manuscript submissions: closed (30 June 2021) | Viewed by 23087

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Prof. Dr. Maurizio Ciani

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Università Politecnica delle Marche, Ancona, Italy
Interests: yeast biotechnology; microbial active compounds; antimicrobial compounds; non-conventional yeasts
Special Issues, Collections and Topics in MDPI journals

Dr. Francesca Comitini

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Dipartimento Scienze della Vita e dell'Ambiente, Università Politecnica delle Marche, Ancona, Italy
Interests: yeast interactions; antimicrobial compounds; fermentation processes; yeast physiology; microbial active compounds
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

In this Special Issue we encourage the submission of papers (research papers or reviews) on the biotechnology of non-conventional yeasts. Non-conventional yeasts have stimulated increasing interest with the development of molecular means that have made it possible to know the genomic profile of several yeast species with promising biotechnological application.

The present Special Issue is focused on recent developments in non-conventional yeasts biotechnology, with topics including recent techniques for characterizing yeasts and their physiology (including omics and nanobiotechnology techniques), methods to adapt industrial strains (including metabolic, synthetic, and evolutionary engineering), and the molecular aspects on the use of yeasts as microbial cell factories to produce biopharmaceuticals, enzymes, alcohols, organic acids, flavors, and fine chemicals. There is a growing interest in the applications of non-conventional yeasts as starters for functional foods or fermented beverages, as well as producers of antimicrobial compounds. In this regard, further investigations on the molecular characteristics of these non-conventional yeasts needed.

Prof. Maurizio Ciani
Dr. Francesca Comitini
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

non-conventional yeasts
yeast biotechnology
omics techniques
nanobiotechnology
industrial yeasts
cell factory
yeast metabolites
antimicrobial compounds
metabolic profile
yeast fermentation

Published Papers (6 papers)

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Research

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20 pages, 1092 KiB

Open AccessArticle

Application of a New Engineered Strain of Yarrowia lipolytica for Effective Production of Calcium Ketoglutarate Dietary Supplements

by Ludwika Tomaszewska-Hetman, Anita Rywińska, Zbigniew Lazar, Piotr Juszczyk, Magdalena Rakicka-Pustułka, Tomasz Janek, Marta Kuźmińska-Bajor and Waldemar Rymowicz

Int. J. Mol. Sci. 2021, 22(14), 7577; https://doi.org/10.3390/ijms22147577 - 15 Jul 2021

Cited by 5 | Viewed by 2318

Abstract

The present study aimed to develop a technology for the production of dietary supplements based on yeast biomass and α-ketoglutaric acid (KGA), produced by a new transformant of Yarrowia lipolytica with improved KGA biosynthesis ability, as well to verify the usefulness of the obtained products for food and feed purposes. Transformants of Y. lipolytica were constructed to overexpress genes encoding glycerol kinase, methylcitrate synthase and mitochondrial organic acid transporter. The strains were compared in terms of growth ability in glycerol- and oil-based media as well as their suitability for KGA biosynthesis in mixed glycerol–oil medium. The impact of different C:N:P ratios on KGA production by selected strain was also evaluated. Application of the strain that overexpressed all three genes in the culture with a C:N:P ratio of 87:5:1 allowed us to obtain 53.1 g/L of KGA with productivity of 0.35 g/Lh and yield of 0.53 g/g. Finally, the possibility of obtaining three different products with desired nutritional and health-beneficial characteristics was demonstrated: (1) calcium α-ketoglutarate (CaKGA) with purity of 89.9% obtained by precipitation of KGA with CaCO₃, (2) yeast biomass with very good nutritional properties, (3) fixed biomass-CaKGA preparation containing 87.2 μg/g of kynurenic acid, which increases the health-promoting value of the product. Full article

(This article belongs to the Special Issue Biotechnology of Non-conventional Yeasts)

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14 pages, 12564 KiB

Open AccessArticle

The Impact of Hanseniaspora vineae Fermentation and Ageing on Lees on the Terpenic Aromatic Profile of White Wines of the Albillo Variety

by Juan Manuel Del Fresno, Carlos Escott, Iris Loira, Francisco Carrau, Rafael Cuerda, Rémi Schneider, María Antonia Bañuelos, Carmen González, José Antonio Suárez-Lepe and Antonio Morata

Int. J. Mol. Sci. 2021, 22(4), 2195; https://doi.org/10.3390/ijms22042195 - 23 Feb 2021

Cited by 14 | Viewed by 2324

Abstract

Hanseniaspora vineae is a non-Saccharomyces yeast that has a powerful impact on the sensory profile of wines. Its effect on the aromatic profile of non-aromatic grape varieties, such as Albillo Mayor (Vitis vinifera, L), during vinification is a useful biotechnology to improve sensory complexity. Fermentation in steel barrels using Hanseniaspora vineae and sequential inoculation with Saccharomyces cerevisiae have been used to study the formation of terpenes and cell lysis in the production of Albillo white wines. The GC-MS analysis profile shows a significant effect of H. vineae fermentation on the contents of terpenes (≈×3), mainly in linalool (>×3), β-citronellol (>×4), geraniol (>×2) and α-terpineol (≈×2). The contents of several polyoxygenated terpenes and some volatile phenols with a spicy aroma were increased during fermentation. In summary, Hanseniaspora vineae releases a large number of cell wall polysaccharides during fermentation that affect wine palatability and structure. Hanseniaspora vineae is a powerful bio-tool to enhance the fruitiness, floral notes and freshness in non-aromatic white varieties. Full article

(This article belongs to the Special Issue Biotechnology of Non-conventional Yeasts)

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16 pages, 1800 KiB

Open AccessArticle

Development of Genetic Modification Tools for Hanseniasporauvarum

by Jennifer Badura, Niël van Wyk, Silvia Brezina, Isak S. Pretorius, Doris Rauhut, Jürgen Wendland and Christian von Wallbrunn

Int. J. Mol. Sci. 2021, 22(4), 1943; https://doi.org/10.3390/ijms22041943 - 16 Feb 2021

Cited by 17 | Viewed by 3141

Abstract

Apiculate yeasts belonging to the genus Hanseniaspora are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies published in recent years. However, tools for their molecular genetic manipulation are still lacking. Here, we report the development of a tool for the genetic modification of Hanseniaspora uvarum. This was employed for the disruption of the HuATF1 gene, which encodes a putative alcohol acetyltransferase involved in acetate ester formation. We generated a synthetic marker gene consisting of the HuTEF1 promoter controlling a hygromycin resistance open reading frame (ORF). This new marker gene was used in disruption cassettes containing long-flanking (1000 bp) homology regions to the target locus. By increasing the antibiotic concentration, transformants were obtained in which both alleles of the putative HuATF1 gene were deleted in a diploid H. uvarum strain. Phenotypic characterisation including fermentation in Müller-Thurgau must showed that the null mutant produced significantly less acetate ester, particularly ethyl acetate. This study marks the first steps in the development of gene modification tools and paves the road for functional gene analyses of this yeast. Full article

(This article belongs to the Special Issue Biotechnology of Non-conventional Yeasts)

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14 pages, 3512 KiB

Open AccessArticle

Yeast Microbiota during Sauerkraut Fermentation and Its Characteristics

by Paweł Satora, Magdalena Skotniczny, Szymon Strnad and Katarína Ženišová

Int. J. Mol. Sci. 2020, 21(24), 9699; https://doi.org/10.3390/ijms21249699 - 18 Dec 2020

Cited by 13 | Viewed by 3356

Abstract

Sauerkraut is the most important fermented vegetable obtained in Europe. It is produced traditionally by spontaneous fermentation of cabbage. The aim of this study was to determine biodiversity of yeasts present during fermentation of eight varieties of cabbages (Ambrosia, Avak, Cabton, Galaxy, Jaguar, Kamienna Głowa, Manama and Ramco), as well as characterize obtained yeast isolates. WL Nutrient Agar with Chloramphenicol was used to enumerate yeast. Isolates were differentiated using RAPD-PCR and identified by sequencing of the 5.8S-ITS rRNA gene region. The volatiles production was analyzed using SPME-GC-TOFMS. Our research confirmed that during sauerkraut fermentation there is an active growth of the yeasts, which begins in the first phases. The maximal number of yeast cells from 1.82 to 4.46 log CFU g⁻¹ occurred after 24 h of fermentation, then decrease in yeast counts was found in all samples. Among the isolates dominated the cultures Debaryomyces hansenii, Clavispora lusitaniae and Rhodotorula mucilaginosa. All isolates could grow at NaCl concentrations higher than 5%, were relatively resistant to low pH and the presence of lactic acid, and most of them were characterized by killer toxins activity. The highest concentration of volatiles (mainly esters and alcohols) were produced by Pichia fermentans and D. hansenii strains. Full article

(This article belongs to the Special Issue Biotechnology of Non-conventional Yeasts)

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Review

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21 pages, 10012 KiB

Open AccessReview

Yeast Interactions and Molecular Mechanisms in Wine Fermentation: A Comprehensive Review

by Francesca Comitini, Alice Agarbati, Laura Canonico and Maurizio Ciani

Int. J. Mol. Sci. 2021, 22(14), 7754; https://doi.org/10.3390/ijms22147754 - 20 Jul 2021

Cited by 34 | Viewed by 5220

Abstract

Wine can be defined as a complex microbial ecosystem, where different microorganisms interact in the function of different biotic and abiotic factors. During natural fermentation, the effect of unpredictable interactions between microorganisms and environmental factors leads to the establishment of a complex and stable microbiota that will define the kinetics of the process and the final product. Controlled multistarter fermentation represents a microbial approach to achieve the dual purpose of having a less risky process and a distinctive final product. Indeed, the interactions evolved between microbial consortium members strongly modulate the final sensorial properties of the wine. Therefore, in well-managed mixed fermentations, the knowledge of molecular mechanisms on the basis of yeast interactions, in a well-defined ecological niche, becomes fundamental to control the winemaking process, representing a tool to achieve such objectives. In the present work, the recent development on the molecular and metabolic interactions between non-Saccharomyces and Saccharomyces yeasts in wine fermentation was reviewed. A particular focus will be reserved on molecular studies regarding the role of nutrients, the production of the main byproducts and volatile compounds, ethanol reduction, and antagonistic actions for biological control in mixed fermentations. Full article

(This article belongs to the Special Issue Biotechnology of Non-conventional Yeasts)

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15 pages, 1417 KiB

Open AccessReview

High Potential of Pichia kluyveri and Other Pichia Species in Wine Technology

by Javier Vicente, Fernando Calderón, Antonio Santos, Domingo Marquina and Santiago Benito

Int. J. Mol. Sci. 2021, 22(3), 1196; https://doi.org/10.3390/ijms22031196 - 26 Jan 2021

Cited by 58 | Viewed by 5655

Abstract

The surfaces of grapes are covered by different yeast species that are important in the first stages of the fermentation process. In recent years, non-Saccharomyces yeasts such as Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, and Pichia kluyveri have become popular with regard to winemaking and improved wine quality. For that reason, several manufacturers started to offer commercially available strains of these non-Saccharomyces species. P. kluyveri stands out, mainly due to its contribution to wine aroma, glycerol, ethanol yield, and killer factor. The metabolism of the yeast allows it to increase volatile molecules such as esters and varietal thiols (aroma-active compounds), which increase the quality of specific varietal wines or neutral ones. It is considered a low- or non-fermentative yeast, so subsequent inoculation of a more fermentative yeast such as Saccharomyces cerevisiae is indispensable to achieve a proper fermented alcohol. The impact of P. kluyveri is not limited to the grape wine industry; it has also been successfully employed in beer, cider, durian, and tequila fermentation, among others, acting as a promising tool in those fermentation processes. Although no Pichia species other than P. kluyveri is available in the regular market, several recent scientific studies show interesting improvements in some wine quality parameters such as aroma, polysaccharides, acid management, and color stability. This could motivate yeast manufacturers to develop products based on those species in the near future. Full article

(This article belongs to the Special Issue Biotechnology of Non-conventional Yeasts)

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