The Membrane Gradostat Reactor: Secondary metabolite production, bioremediation and commercial potential

This manuscript focuses on the aspect of a membrane gradostat as an entirely different concept compared to submerged hollow fibre modules. The use of membrane bioreactor (MBR) technology is rapidly advancing in the wastewater treatment industries. However, this is not the case in the biopharmaceutical manufacturing industries. The MGR has shown great potential and versatility in terms of industrial applications. It can be used in both wastewater treatment and biopharmaceutical manufacturing using different modes of operation to meet any predetermined process  requirements. The MGR concept uses capillary membranes, which contain microvoids in the substructure to immobilise microbial cells or enzymes, depending on the bioreactor’s application. Operational requirements of the MGR and its commercial potential are discussed from a bioprocess engineering perspective

Malic Acid in Wine: Origin, Function and Metabolism during Vinification

The production of quality wines requires a judicious balance between the sugar, acid and flavour components of wine.  L-Malic and tartaric acids are the most prominent organic acids in wine and play a crucial role in the winemaking process, including the organoleptic quality and the physical, biochemical and microbial stability of wine.  Deacidification of grape must and wine is often required for the production of well-balanced wines. Malolactic fermentation induced by the addition of malolactic starter cultures, regarded as the preferred method for naturally reducing wine acidity, efficiently decreases the acidic taste of wine, improves the microbial stability and modifies to some extent the organoleptic character of wine. However, the recurrent phenomenon of delayed or sluggish malolactic fermentation often causes interruption of cellar operations, while the malolactic fermentation is not always compatible with certain styles of wine. Commercial wine yeast strains of Saccharomyces are generally unableto degrade L-malic acid effectively in grape must during alcoholic fermentation, with relatively minor modifications in total acidity during vinification. Functional expression of the malolactic pathway genes, i.e. the malate transporter (mae1) of Schizosaccharomyces pombe and the malolactic enzyme (mleA) from Oenococcus oeni in wine yeasts, haspaved the way for the construction of malate-degrading strains of Saccharomyces for commercial winemaking

The winemaker’s bug: From ancient wisdom to opening new vistas with frontier yeast science

The past three decades have seen a global wine glut. So far, well-intended but wasteful and expensive market-intervention has failed to drag the wine industry out of a chronic annual oversupply of roughly 15%. Can yeast research succeed where these approaches have failed by providing a means of improving wine quality, thereby making wine more appealing to consumers? To molecular biologists Saccharomyces cerevisiae is as intriguing as it is tractable. A simple unicellular eukaryote, it is an ideal model organism, enabling scientists to shed new light on some of the biggest scientific challenges such as the biology of cancer and aging. It is amenable to almost any modification that modern biology can throw at a cell, making it an ideal host for genetic manipulation, whether by the application of traditional or modern genetic techniques. To the winemaker, this yeast is integral to crafting wonderful, complex wines from simple, sugar-rich grape juice. Thus any improvements that we can make to wine, yeast fermentation performance or the sensory properties it imparts to wine will benefit winemakers and consumers. With this in mind, the application of frontier technologies, particularly the burgeoning fields of systems and synthetic biology, have much to offer in their pursuit of “novel” yeast strains to produce high quality wine. This paper discusses the nexus between yeast research and winemaking. It also addresses how winemakers and scientists face up to the challenges of consumer perceptions and opinions regarding the intervention of science and technology; the greater this intervention, the stronger the criticism that wine is no longer “natural.” How can wine researchers respond to the growing number of wine commentators and consumers who feel that scientific endeavors favor wine quantity over quality and “technical sophistication, fermentation reliability and product consistency” over “artisanal variation”? This paper seeks to present yeast research in a new light and a new context, and it raises important questions about the direction of yeast research, its contribution to science and the future of winemaking

Malic Acid Distribution and Degradation in Grape Must During Skin Contact: The Influence of Recombinant Malo-Ethanolic Wine Yeast Strains

Wine acidity plays an important role in determining wine quality and ensuring physiochemical and microbiological stability. In high-acid wines, the L-malic acid concentration is usually reduced through bacterial malolactic fermentation, while acidulation in low-acidity wines is usually done during final blending of the wine before bottling.  This study showed that skin contact did not influence the relative concentration of L-malic acid in the pulp and juice fractions from Colombard, Ruby Cabernet and Cabernet Sauvignon grape musts, with 32%-44% of the L-malic acid present in the pulp fraction. Four recombinant malo-ethanolic (ME) Saccharomyces wine yeast strains containing the malic enzyme (mae2) and malate transporter (mael) genes of Schizasaccharomyces pombe, effectively degraded the L-malic acid in both the juice and pulp fractions of all three cultivars, with a complete degradation of malic acid in the juice fraction within 2 days

Genetic Engineering of an Industrial Strain of Saccharomyces cerevisiae for L-Malic Acid Degradation via an Efficient Malo-Ethanolic Pathway

The optimal ratio of L-malic and L-tartaric acid in relation to other wine components is one of the most important aspects that ultimately determine wine quality during winemaking. Winemakers routinely rely on the judicious use of malolactic fermentation (MLF) after alcoholic fermentation to deacidify and stabilise their wines. However, due to theunreliability of the process and unsuitable sensory modifications in some grape cultivars, especially for fruity-floral wines, MLF is often regarded as problematic and undesirable. Alternative methods for reducing the amounts of L-malic acid in wine will contribute to improving the production of quality wines in the future, especially in coolclimate regions. Most wine yeast strains of Saccharomyces are unable to effectively degrade L-malic acid, whereas the fission yeast Schizosaccharomyces pombe efficiently degrades high concentrations of L-malic acid by means of malo-ethanolic fermentation. However, strains of S. pombe are not suitable for vinification due to the production of undesirable off-flavours. Previously, the 5. pombe malate permease (mael) and malic enzyme (mae2) genes were  successfully expressed under the 3-phosphoglycerate kinase (PGK1) regulatory elements in 5. cerevisiae, resulting in a recombinant laboratory strain of S. cerevisiae with an efficient malo-ethanolic pathway. Stable integration of the S. pombe malo-ethanolic pathway genes has now been obtained through the construction of a unique integration strategy in a commercial wine yeast strain. Co-transformation of the linear integration cassette containing the mael and mae2 genes and PGK1 regulatory elements and a multi-copy plasmid containing the phleomycin-resistance marker into a commercial Saccharomyces cerevisiae strain resulted in the successful transformation and integration of the malo-ethanolic genes. The recombinant 5. cerevisiae strain was successfully cured of phleomycin-resistance plasmid DNA in order to obtain malo-ethanolic yeast containing only yeast-derived DNA. The integrated malo-ethanolic genes were stable in 5. cerevisiae and during synthetic and grape must fermentation, L-malic acid was completely fermented to ethanol without any negative effect on fermentation kinetics and wine quality

Berry size variation of vitis vinifera l. Cv. Syrah : morphological dimensions, berry composition and wine quality

CITATION: Melo, M. S., et al. 2015. Berry size variation of vitis vinifera l. Cv. Syrah : morphological dimensions, berry composition and wine quality. South African Journal for Enology and Viticulture, 36(1):1-10, doi:10.21548/36-1-931.The original publication is available at http://www.journals.ac.za/index.php/sajevBerry size has always been a quality factor in wine production. In this study, Syrah grapes from a single vineyard were classified into different size groups according to diameter: small ( 14 mm). Smaller berries were present in the highest and larger berries in the lowest numbers. Size distributions were similar in both seasons (2010/2011 and 2011/2012). Berry physical characteristics (mass, volume and skin area) increased with size, showing the same tendency in both years. Positive correlations between berry mass, volume and skin area were found, whereas these variables were negatively related with berry number/kg grapes. Berry volume was negatively correlated with dry skin weight. Skin surface area/berry volume seems to be an indicator of the “dilution” effect associated with increasing size, as larger berries presented the lowest values. In 2012 the grapes were harvested at a higher soluble solid level than in the previous year; large-sized berries presented the lowest levels in both years. The whole-berry analysis of total anthocyanins showed a decrease in concentration and increase in content per berry, from smaller to larger berries. Small berries and the control (naturally occurring berry size mixture) showed a higher extractability of anthocyanins and phenolic compounds than the medium and large berries. Sensorially, wines from medium berries were more consistent over the two years, scoring higher than the rest. Berry sizes were related to wine style differences, and knowing the population of berry sizes in the vineyard close to harvest would offer a possibility to predict wine styles.http://www.journals.ac.za/index.php/sajev/article/view/931Publisher's versio

Qualitative and quantitative detection of SARS-CoV-2 RNA in untreated wastewater in Western Cape Province, South Africa

Recent studies have shown that the detection of SARS-CoV-2 genetic material in wastewater may provide the basis for a surveillance system to track the environmental dissemination of this virus in communities. An effective wastewater-based epidemiology (WBE) system may prove critical in South Africa (SA), where health systems infrastructure, testing capacity, personal protective equipment and human resource capacity are constrained. In this proof-of-concept study, we investigated the potential of SARS-CoV-2 RNA surveillance in untreated wastewater as the basis for a system to monitor COVID-19 prevalence in the population, an early warning system for increased transmission, and a monitoring system to assess the effectiveness of interventions. The laboratory confirmed the presence (qualitative analysis) and determined the RNA copy number of SARS-CoV-2 viral RNA by reverse transcription polymerase chain reaction (quantitative) analysis from 24-hour composite samples collected on 18 June 2020 from five wastewater treatment plants in Western Cape Province, SA. The study has shown that a WBE system for monitoring the status and trends of COVID-19 mass infection in SA is viable, and its development and implementation may facilitate the rapid identification of hotspots for evidence-informed interventions

Phenotypic and genotypic diversity of wine yeasts used for acidic musts

The aim of this study was to examine the physiological and genetic stability of the industrial wine yeasts Saccharomyces cerevisiae and Saccharomyces bayanus var. uvarum under acidic stress during fermentation. The yeasts were sub-cultured in aerobic or fermentative conditions in media with or without l-malic acid. Changes in the biochemical profiles, karyotypes, and mitochondrial DNA profiles were assessed after minimum 50 generations. All yeast segregates showed a tendency to increase the range of compounds used as sole carbon sources. The wild strains and their segregates were aneuploidal or diploidal. One of the four strains of S. cerevisiae did not reveal any changes in the electrophoretic profiles of chromosomal and mitochondrial DNA, irrespective of culture conditions. The extent of genomic changes in the other yeasts was strain-dependent. In the karyotypes of the segregates, the loss of up to 2 and the appearance up to 3 bands was noted. The changes in their mtDNA patterns were much broader, reaching 5 missing and 10 additional bands. The only exception was S. bayanus var. uvarum Y.00779, characterized by significantly greater genome plasticity only under fermentative stress. Changes in karyotypes and mtDNA profiles prove that fermentative stress is the main driving force of the adaptive evolution of the yeasts. l-malic acid does not influence the extent of genomic changes and the resistance of wine yeasts exhibiting increased demalication activity to acidic stress is rather related to their ability to decompose this acid. The phenotypic changes in segregates, which were found even in yeasts that did not reveal deviations in their DNA profiles, show that phenotypic characterization may be misleading in wine yeast identification. Because of yeast gross genomic diversity, karyotyping even though it does not seem to be a good discriminative tool, can be useful in determining the stability of wine yeasts. Restriction analysis of mitochondrial DNA appears to be a more sensitive method allowing for an early detection of genotypic changes in yeasts. Thus, if both of these methods are applied, it is possible to conduct the quick routine assessment of wine yeast stability in pure culture collections depositing industrial strains

Factors influencing radiography lecturers’ perceptions and understanding of reflective practice in a newly implemented curriculum

Background. Reflective practice has become an integral component of the new Bachelor of Science (BSc) radiography programme in South Africa (SA). As a result, lecturers on the programme are required to facilitate reflective learning, and are assumed to be skilled in this. However, relevant literature indicates that health professions educators may not necessarily possess the requisite competence or experience in facilitating reflective learning. Moreover, there is a paucity in the literature on lecturers’ perceptions and understanding of reflective practice in the context of undergraduate radiography curricula, particularly in sub-Saharan Africa.Objective. To gain insight into BSc radiography lecturers’ perceptions and understanding of reflective practice at a selected university of technology in SA.Methods. This was a qualitative exploratory study in which individual semi-structured interviews were conducted with lecturers teaching on the new BSc Radiography programme. The purposively selected sample consisted of 11 participants. Interviews were audio-recorded, transcribed and thematically analysed.Results. Analysis of the data revealed three themes, namely diverse understandings of reflective practice among lecturers, factors  influencing optimal facilitation of reflective learning and strategies to improve reflective practice.Conclusion. The findings of this study indicated that lecturers felt unprepared to facilitate reflective practice in the new BSc radiography curriculum. The need for faculty development initiatives, such as an introduction to reflective tools and educational strategies to support lecturers in facilitating reflective practice, was highlighted. Furthermore, it was found that if objectives were clearly outlined and facilitator guides available, a desired reflective practice could be establishe

Oxygen mass transfer for an immobilised biofilm of Phanerochaete chrysosporium in a membrane gradostat bioreactor.

A novel system, the membrane gradostat reactor (MGR), designed for the continuous production of secondary metabolites, has been shown to have higher production per reactor volume than batch culture systems. The MGR system mimics the natural environment in which wild occurring microorganism biofilms flourish. The biofilms are immobilised on the external surface of an ultrafiltration membrane where substrate distribution gradients are established across the biofilm. The hypothesis that, dissolved oxygen (DO) mass transfer parameters obtained in submerged pellets can be used to describe and model DO mass transfer parameters in the MGR, was refuted. Phanerochaete chrysosporium biofilms, immobilised on ultrafiltration capillary membranes in the MGR systems were used to quantify DO distribution using a Clark-type microsensor. The DO penetration depth decreased with increasing biofilm thickness, which resulted in the formation of anaerobic zones in the biofilms. Oxygen flux values of 0.27 to 0.7 g/(m2.h) were obtained during the MGR operation. The consumption of oxygen and the Monod saturation constants used in the modelling of oxygen distribution in immobilised biofilms were in the range of 894.53 to 2739.70 g/(m3.h) and 0.041 to 0.999 g/m3, respectivel