Exploring the phenotypic diversity of oenological traits in Kluyveromyces marxianus strains

Date
2021-12
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Stellenbosch : Stellenbosch University
Abstract
ENGLISH ABSTRACT: A wide range of yeast species occur in the wine environment. While Saccharomyces cerevisiae dominates alcoholic fermentations, other species may also confer positive organoleptic attributes. Although more frequently isolated from the dairy, cocoa and tequila environments and highly studied for its biotechnological potential, Kluyveromyces marxianus has been occasionally reported in the wine environment. Recent reports have demonstrated that strain IWBT Y885, isolated from South African grape must, displays promising properties for winemaking which includes its high pectinase activity and positive contributions to aroma production. Like other yeast species, a large degree of intraspecific variability has been shown to occur within the species. This study aimed to investigate and compare five different K. marxianus strains (L01 to L05) isolated from diverse substrates (dairy, beer, distillery) with strain IWBT Y885, with the underlying goal of interrogating whether the relevant winemaking characteristics previously reported in the latter strain are unique or common amongst K. marxianus strains. Along with these strains, the newly produced fusant BF2020 was also investigated. This fusant was generated as a hybrid product of a wine strain of S. cerevisiae and K. marxianus IWBT Y885 and displays intermediate fermentative capabilities. Firstly, the morphological traits of the strains investigated were compared. Various differences could be identified with only a select few displaying similar morphologies to IWBT Y885. Subsequently, various oenological traits were investigated in synthetic grape juice. These included fermentation performance, fermentation metabolites, hydrogen sulfide and pectinase production. Variations between strain IWBT Y885 and strains L01 through L05 were found for the majority of traits investigated, regardless of their source of isolation. Following Cluster analysis, three strain groupings emerged. The first group comprised strains L01, L05, Y885 and BF2020 and the second group strains L02 and L04. Strain L03 appeared to show the most variation from the other strains and clustered separately. Thereafter, fermentations were completed in Chenin blanc as well as Cabernet Sauvignon grape musts under conditions simulating those of industrial settings. Using cluster analysis of the major volatiles produced, clear separations could be observed between all sequential Chenin blanc K. marxianus fermentations and the S. cerevisiae fermentations. Similar groupings previously observed in synthetic grape juice emerged between the K. marxianus strains. However, these chemical differences did not translate into clear sensory differences. It is hypothesized that potential differences in yeast-derived aroma compounds may have been overshadowed by more dominant sensory attributes (fresh green, chalky, bitter, fizzy) listed by the sensory panel. This study confirmed that strain variation is a common occurrence within K. marxianus. In a winemaking context, it leads to the production of varying concentrations of aroma compounds. Subsequently, strain IWBT Y885 stands out with regards to various oenological factors when investigated in synthetic grape juice, however real grape juice comparisons require further investigations.
AFRIKAANSE OPSOMMING: ‘n Groot verskeidenheid van gis-spesies kom voor in die wyn-ongewing. Terwyl Saccharomyces cerevisiae alkoholiese fermentasie domineer, is dit ook waar dat ander spesies ‘n positiewe invloed op die organoleptiese eienskappe van wyn kan uitoefen. Alhoewel K. marxianus meestal in suiwel, kakao en tequila omgewings gevind word, word hierdie gis ook gevind in verskeie wyn-omgewings. Onlangse studies demonstreer dat die ras IWBT Y885, wat van Suid-Afrikaanse druiwesap geisoleer was, baie belowende eienskappe vir die wynbedryf inhou. Hierdie sluit in hoë pektinase aktiwiteite sowel as ‘n positiewe bydra tot die aroma produksie. Soos met ander gis-spesies, is daar ‘n groot hoeveelheid van intraspesifieke verskille tussen K. marxianus rasse. Die hoofdoel van hierdie studie was om vyf verskillende K. marxianus rasse (L01 tot L05), wat van verskeie omgewings geisoleer was (suiwel, bier, distilleerdery), met die ras IWBT Y885 te vergelyk. Hierdie vergelyking fokus spesifiek op die relevante wynmaak-einskappe wat reeds in Y885 gevind word. Saam met hierdie rasse was die nuut-gemaakde “fusant” gis BF2020 ingesluit. BF2020 was gemaak as ‘n produk van die wyn-gis S. cerevisiae en K. marxianus Y885. Eerstens was die morfologiese eienskappe van die rasse ondersoek en vergelyk. Talle verskille was gevind tussen die giste met net sommige giste wat dieselfde morfologiese eienskappe gewys het as IWBT Y885. Daarna was verskeie wynkundige eienskappe ondersoek in kuns druiwesap. Hierdie het bestaan uit fermentasie krag, fermentasie metaboliese produkte, sowel as H2S en pektinase produksie. Verskille tussen Y885 en die rasse L01 tot L05 was gevind in die meederheid van eienskappe wat ondersoek is. Hierdie verskille het voorgekom ten spyte van die oorsprong van die giste. Nadat ‘n cluster analiese gedoen was, het drie groeperings verskyn. Die eerste groep het uit rasse L01, L05, Y885 en BF2020 bestaan en die tweede groep uit L02 en L04. L03 was die mees verskillend van die ander rasse en het die derde groep gemaak. Na die eksperimente in kuns druiwesap was fermentasies gedoen met Chenin blanc en Cabernet Sauvignon druiwesap. Deur cluster analiese te gebruik vir die vlugtige stowwe het duidelike verskille geblyk tussen die Chenin blanc fermentasies met K. marxianus en dié met S. cerevisiae. Soortgelyke groeperings het verskyn tussen die K. marxianus rasse soos wat gesien was in die kuns druiwesap fermentasies. Ten spyte van die chemiese verskille tussen die wyne, was geen duidelike sensoriese verskille gevind nie. Die hipotese is dat die moonlike verskille wat verskyn het, nie opgetel was nie as gevolg van die meer dominante sensoriese eienskappe wat in die wyn gevind was (vars groen, kalkerig, bitter, bruisend). Hierdie studie het bevestig dat verskille tussen K. marxianus rasse ‘n algemene voorkoms is. Spesifiek in die wynbedryf laat dit toe dat wyne met verskillende chemiese komposisies gemaak kan word. Die ras IWBT Y885 het duidelik uitgestaan met verwysing na verskeie wynkundige eienskappe in kuns druiwesap, maar hierdie eienskappe benodig verdere ondersoek met verband na regte druiwesap eksperimente.
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Thesis (MScAgric)--Stellenbosch University, 2021.
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