Industrial Baker Research Articles

Enhancement of fermentation traits in industrial Baker's yeast for low or high sugar environments

Saccharomyces cerevisiae SPC-SNU 70-1 is a commercial diploid baking yeast strain valued for its excellent bread-making qualities, including superior leavening capabilities and the production of flavor-enhancing volatile organic acids. Despite its benefits, this strain faces challenges in fermenting both lean (low-sugar) and sweet (high-sugar) doughs. To address these issues, we employed the CRISPR/Cas9 genome editing system to modify genes without leaving any genetic scars. For lean doughs, we enhanced the yeast's ability to utilize maltose over glucose by deleting a gene involved in glucose repression. For sweet doughs, we increased glycerol production by overexpressing glycerol biosynthetic genes and optimizing redox balance, thereby improving the tolerence to osmotic stress during fermentation. Additionally, the glycerol-overproducing strain demonstrated enhanced freeze tolerance, and bread made from this strain exhibited improved storage properties. This study demonstrates the feasibility and benefits of using engineered yeast strains, created solely by editing their own genes without introducing foreign genes, to enhance bread making.

Specific antioxidant enzymes are involved in the freeze-thawing response of industrial baker's yeasts.

In this study, the biochemical basis of resistance to slow freezing and thawing (F-T) stress was explored in two baker yeast industrial strains of Saccharomyces cerevisiae that presented differential tolerance to freezing in order to be in the frozen bakery industry. Strain Y8, used commercially in sweet baking doughs, exhibited greater stress tolerance than Y9, a strain employed in regular doughs. Survival of Y8 was higher than that of Y9 (30%vs 12%) after F-T or other reactive oxygen species (ROS) inducing stresses compared to their non-stressed controls. The superior F-T tolerance of Y8 was related to its lower ROS accumulation capacity, determined by fluorometry in cell-free extracts and in vivo, by fluorescence microscopy upon F-T, being Y8 ROS accumulation 2-fold lower than that of Y9. That, in turn, could be positively associated with Y8's higher constitutive activities of cytosolic catalase (CAT) and superoxide dismutase by a significant activation (25%) of Y8 CAT after F-T. That would complement the protective effects of other protectant molecules like trehalose, present at high concentration in this strain.

Conversion of Problematic Winery Waste into Valuable Substrate for Baker's Yeast Production and Solid Biofuel: A Circular Economy Approach§.

Wine production, which is considered a major sector in food industry, often involves the use of a large amount of resources. Moreover, wine making generates a large amount of grape pomace, which is generally used for low-value applications such as fertiliser and animal feed. The aim of the present research is to explore the possibility of improving the overall sustainability of traditional winemaking. A zero-waste process was developed. It includes the production of white wine and the substantial valorisation of grape pomace, which is converted into solid biofuel, tartaric acid and concentrated grape extract as feedstock for industrial baker's yeast production. We estimate that a significant surplus of renewable energy of approx. 3 MJ/kg processed grapes can be obtained during this conversion. The suitability of grape extract as a potential substrate for industrial baker's yeast production was evaluated and the feasibility of a partial replacement of molasses (up to 30 %) was demonstrated. We present a circular economy approach for the conversion of winery biowaste into high-value resources such as feedstock and solid biofuel.

The impacts of nicotinamide and inositol on the available cells and product performance of industrial baker's yeasts

A suitable nutrient supply, especially of vitamins, is very significant for the deep display of the inherent genetic properties of microorganisms. Here, using the chemically defined minimal medium (MM) for yeast, nicotinamide and inositol were confirmed to be more beneficial for the performance of two industrial baker's yeasts, a conventional and a high-sugar-tolerant strain. Increasing nicotinamide or inositol to proper levels could enhance the both strains on cell growth and activity and product performance, including trehalose accumulation and leavening performance. The activity of key enzymes (PCK, TPS) and the content of intermediate metabolites (G6P, UDPG) in the trehalose synthesis pathway were promoted by a moderate supply of nicotinamide and inositol. That were also proved that an appropriate amount of niacinamide promoted the transcription of longevity-related genes (PNC1, SIR2), and the proper concentration of inositol altered the phospholipid composition in cells, namely, phosphatidylinositol and phosphatidyl choline. Furthermore, the cell growth and the leavening performance of the both strains were promoted after adjusting inositol to choline to the proper ratio, resulting directly in content changes of phosphatidylinositol and phosphatidyl choline in the cells. While the two strains responded to the different proper ratio of inositol to choline probably due to their specific physiological characteristics. Such beneficial effects of increased nicotinamide levels were confirmed in natural media, molasses and corn starch hydrolyzed sugar media. Meanwhile, such adjustment of inositol to choline ratio could lessen the inhibition of excess inositol on cell growth of the two tested strains in corn starch hydrolyzed sugar media. However, in molasse, such phenomenon was not observed probably since there was higher Ca2+ in it. The results indicated that the effects of nutrient factors, such as vitamins, on cell growth and other properties found out from the simple chemically defined minimal medium were an effective measure to use in improving the recipe of natural media at least for baker's yeast.

Oxidative stability of cocoa hazelnut cream enriched with inactive yeast cells

The aim of the study was to evaluate the effect of inactivated yeast cells on the oxidative stability, color, and sensory properties of cocoa hazelnut cream. For this purpose, yeast cells were inactivated and added in the cocoa hazelnut cream. Glutathione content, antioxidant properties, TBARS, color, and sensory properties of the samples were examined during 150 days. According to the results, the GSH content in the cocoa hazelnut cream after yeast addition was 47.55 nmol GSH/g sample. In addition, TBARS value was lower in yeast-added sample (0.1846 mg MDA/kg sample) compared to control (0.2262 mg MDA/kg sample) at the end of storage. Moreover, the DPPH values of the hazelnut cream did not change with the yeast addition, significantly. However, yeast-added samples had higher FRAP, CUPRAC, and total phenolic content than the control sample. Sensory scores of yeast-added samples were also acceptable compared to control samples. Practical applications In this study, the effect of inactive industrial baker's yeast (Saccharomyces cerevisiae) addition on the functional and sensory properties of the cocoa hazelnut cream was investigated. The results have shown that industrial baker's yeast can be encapsulated by heat treatment and helps to maintain oxidative stability of cocoa hazelnut cream without causing significant changes in the sensory properties during storage. Thus, the addition of inactivate yeast cells can be recommended for the further development of novel and innovative nutritional products with natural ingredients.

Full Mass Balance Analysis During Industrial Baker's Yeast Fermentation Shows New Perspectives for Biomolecule Recovery

Abstract This study aims to partially compensate the lack of reliable data regarding full mass balance analysis during industrial baker's yeast fermentation and the chemical composition of the resu...

Construction of industrial baker's yeast with high level of cAMP.

Cyclic adenosine monophosphate (cAMP) plays an important role in modulating the activity of microbe cell. In this study, PKA (protein kinase A) activity was weakened through truncation of TPK2 promoter (-150bp and -300bp) and gene deletion of BCY1 (encodes the regulatory subunit of PKA), TPK1 and TPK3, generating strains BY9a-T2-150 and BY9a-T2-300, respectively. High-performance liquid chromatography analysis showed cAMP levels in BY9a-T2-150 and BY9a-T2-300 were increased by 5- and 18-fold, respectively, compared with that of parent strain, BY9a. The expression levels of TPK2 gene in two engineered strains were decreased by 95% and 97% compared with that of BY9a, respectively. The PKA activity reflected by heat resistance of engineered strains enhanced compared with parent strain BY9a. This study show a new method to increase the intracellular cAMP concentration in industrial yeast by fine-tuning of PKA activity, without influence in growth and fermentation properties. PRACTICAL APPLICATIONS: cAMP as the "second messenger," is essential for plant, animal, and microorganisms and human life. But its synthesis is still limited by expensive cost and time-consuming method. We constructed the industrial baker's yeast with high level of cAMP and desired to be used to produce functional food for relaxing smooth muscle, expanding blood vessels, improving liver function, and promoting nerve regeneration and as a food additive for treating hyperthyreosis and hepatopathy. The methods of two step homologous recombination and backcross operated in this study eliminate the exogenous gene in engineered strains, made it safety to be used in food production. Fine-tuning of PKA activity in engineered strains ensure produce high level of cAMP and exhibit normal growth performance in engineering strains. Therefore, this work is significantin functional foods product and has the potential to be used in practical application.

An Innovative Biocatalyst for Continuous 2G Ethanol Production from Xylo-Oligomers by Saccharomyces cerevisiae through Simultaneous Hydrolysis, Isomerization, and Fermentation (SHIF)

Many approaches have been considered aimed at ethanol production from the hemicellulosic fraction of biomass. However, the industrial implementation of this process has been hindered by some bottlenecks, one of the most important being the ease of contamination of the bioreactor by bacteria that metabolize xylose. This work focuses on overcoming this problem through the fermentation of xylulose (the xylose isomer) by native Saccharomyces cerevisiae using xylo-oligomers as substrate. A new concept of biocatalyst is proposed, containing xylanases and xylose isomerase (XI) covalently immobilized on chitosan, and co-encapsulated with industrial baker’s yeast in Ca-alginate gel spherical particles. Xylo-oligomers are hydrolyzed, xylose is isomerized, and finally xylulose is fermented to ethanol, all taking place simultaneously, in a process called simultaneous hydrolysis, isomerization, and fermentation (SHIF). Among several tested xylanases, Multifect CX XL A03139 was selected to compose the biocatalyst bead. Influences of pH, Ca2+, and Mg2+ concentrations on the isomerization step were assessed. Experiments of SHIF using birchwood xylan resulted in an ethanol yield of 0.39 g/g, (76% of the theoretical), selectivity of 3.12 gethanol/gxylitol, and ethanol productivity of 0.26 g/L/h.

Turning industrial aerobic fermentation plants into thermal power stations

The industrial aerobic bioprocess of baker's yeast production requires large amounts of water to cool down large bubble column bioreactors. As result, an appreciable quantity of low-grade heat is generated. Because of the low temperature level of the water (~25°C) exiting the bioreactors cooling system, very little attention has been dedicated to heat recovery and conversion from this stream, which is usually released in rivers, streams, and canals. In this work, we simulated the generation of low-grade heat (up to 14.4 MW) from an industrial baker's yeast production plant consisting of seven 150-m3 bioreactors. Subsequently, a dedicated transcritical carbon dioxide heat pump system for the conversion of this low-grade heat into fourth generation district heat (~16.2 MW) was successfully designed. Fourth generation district heat employs low-temperature water (30-70°C) as heat carrier and is expected to play a major role in future sustainable energy system. Finally, an economic study confirmed the feasibility and the applicability of our approach and a concept for long-term energy storage including state-of-the-art phase change material (PCM)–units was discussed.

New trends in industrial baker's yeast fermentation: Recovery of key biomolecules and low-grade heat conversion

New trends in industrial baker's yeast fermentation: Recovery of key biomolecules and low-grade heat conversion

Selection and Characterization of Potential Baker's Yeast from Indigenous Resources of Nepal.

The study aims to isolate the yeast strains that could be used effectively as baker's yeast and compare them with the commercial baker's yeast available in the market of Nepal. A total of 10 samples including locally available sources like fruits, Murcha, and a local tree “Dar” were collected from different localities of Bhaktapur, Kavre, and Syangja districts of Nepal, respectively. Following enrichment and fermentation of the samples, 26 yeast strains were isolated using selective medium Wallerstein Laboratory Nutrient Agar. From the differential tests which included morphological and microscopic observation and physiological and biochemical characterization such as nitrate reduction and lactose utilization tests, 8 strains were selected as possible Saccharomyces strain. The selected strains were further assessed for their efficient leavening ability by tests such as ethanol tolerance, osmotolerance, invertase test, and stress exclusion test. The three most potent strains ENG, MUR3B, and SUG1 isolated from grape, Murcha, and sugarcane, respectively, were used in the fermentation and baking of dough. These strains also carried a possibility of being used as industrial baker's yeast.

Functional analysis of the global repressor Tup1 for maltose metabolism in Saccharomyces cerevisiae: different roles of the functional domains

BackgroundTup1 is a general transcriptional repressor of diverse gene families coordinately controlled by glucose repression, mating type, and other mechanisms in Saccharomyces cerevisiae. Several functional domains of Tup1 have been identified, each of which has differing effects on transcriptional repression. In this study, we aim to investigate the role of Tup1 and its domains in maltose metabolism of industrial baker’s yeast. To this end, a battery of in-frame truncations in the TUP1 gene coding region were performed in the industrial baker’s yeasts with different genetic background, and the maltose metabolism, leavening ability, MAL gene expression levels, and growth characteristics were investigated.ResultsThe results suggest that the TUP1 gene is essential to maltose metabolism in industrial baker’s yeast. Importantly, different domains of Tup1 play different roles in glucose repression and maltose metabolism of industrial baker’s yeast cells. The Ssn6 interaction, N-terminal repression and C-terminal repression domains might play roles in the regulation of MAL transcription by Tup1 for maltose metabolism of baker’s yeast. The WD region lacking the first repeat could influence the regulation of maltose metabolism directly, rather than indirectly through glucose repression.ConclusionsThese findings lay a foundation for the optimization of industrial baker’s yeast strains for accelerated maltose metabolism and facilitate future research on glucose repression in other sugar metabolism.

Industrial baker's yeast fermentation: From manufacture to integrated sustainability

A series of novel L-malic acid modified hyper-cross-linked resins were synthesized from macroporous chloromethylated styrene-divinylbenzene copolymer by Fridel–Crafts reaction and esterification reaction. The adsorption behavior of the resin toward p-aminobenzoic acid was investigated from aqueous solution. KZ-3 displayed the most outstanding adsorption capacity due to its proper surface structure and higher functional group utilization rate. Subsequently, the adsorption kinetic of p-aminobenzoic acid onto precursor resin Z-3, modified resin KZ-3 and commercially available polymeric adsorbent H-103 was compared. The result demonstrated that KZ-3 had the optimal adsorption capacity because of its hydrogen bonding driving force and hydrophilicity enhancement which brought by oxygen-containing functional groups. Especially, KZ-3 is 3 times higher than the precursor resin in terms of adsorption rate. Kinetic curves could be characterized by pseudo-second-order rate equation and both the intra-particle diffusion and the film diffusion were the rate-limiting steps. The isotherms could be fitted by Langmuir model and the adsorption was an exothermic, spontaneous and more ordered arrangement process due to its negative thermodynamic parameters.

Impact of Saccharomyces cerevisiae metabolites produced during fermentation on bread quality parameters: A review

ABSTRACTAlthough bread making with the use of Baker's yeast has a long tradition in human history, little attention has been paid to the connection between yeast addition and the final bread quality. Nowadays, bakers mainly use different flour additives such as enzymes (amylases, hemicellulases, and proteases) to change and improve dough properties and/or bread quality. Another strategy is the use of modified industrial Baker's yeast. To date, there is no yeast strain used in the baking industry, which is genetically modified, despite some studies demonstrating that the application of recombinant DNA technology is a possibility for improved strains suitable for baking. However, due to the fact that the majority of consumers in Europe highly reject the use of genetically modified microorganisms in the production of food, other strategies to improve bread quality must be investigated. Such a strategy would be a reconsideration of the selection of yeast strains used for the baking process. Next to the common criteria, the requirement for adequate gas production, more attention should be paid on how yeast impacts flavor, shelf life, color, and the nutritional value of baked products, in a similar way to which yeast strains are selected in the wine and brewing industries.

Development of intra-strain self-cloning procedure for breeding baker's yeast strains

Previously reported self-cloning procedures for breeding of industrial yeast strains require DNA from other strains, plasmid DNA, or mutagenesis. Therefore, we aimed to construct a self-cloning baker's yeast strain that exhibits freeze tolerance via an improved self-cloning procedure. We first disrupted the URA3 gene of a prototrophic baker's yeast strain without the use of any marker gene, resulting in a Δura3 homozygous disruptant. Then, the URA3 gene of the parental baker's yeast strain was used as a selection marker to introduce the constitutive TDH3 promoter upstream of the PDE2 gene encoding high-affinity cyclic AMP phosphodiesterase. This self-cloning procedure was performed without using DNA from other Saccharomyces cerevisiae strains, plasmid DNA, or mutagenesis and was therefore designated an intra-strain self-cloning procedure. Using this self-cloning procedure, we succeeded in producing self-cloning baker's yeast strains that harbor the TDH3p-PDE2 gene heterozygously and homozygously, designated TDH3p-PDE2 hetero and TDH3p-PDE2 homo strains, respectively. These self-cloning strains expressed much higher levels of PDE2 mRNA than the parental strain and exhibited higher viability after freeze stress, as well as higher fermentation ability in frozen dough, when compared with the parental strain. The TDH3p-PDE2 homo strain was genetically more stable than the TDH3p-PDE2 hetero strain. These results indicate that both heterozygous and homozygous strains of self-cloning PDE2-overexpressing freeze-tolerant strains of industrial baker's yeast can be prepared using the intra-strain self-cloning procedure,and, from a practical viewpoint, the TDH3p-PDE2 homo strain constructed in this study is preferable to the TDH3p-PDE2 hetero strain for frozen dough baking.

Enhanced leavening properties of baker's yeast by reducing sucrase activity in sweet dough.

Leavening ability in sweet dough is required for the commercial applications of baker's yeast. This property depends on many factors, such as glycolytic activity, sucrase activity, and osmotolerance. This study explored the importance of sucrase level on the leavening ability of baker's yeast in sweet dough. Furthermore, the baker's yeast strains with varying sucrase activities were constructed by deleting SUC2, which encodes sucrase or replacing the SUC2 promoter with the VPS8/TEF1 promoter. The results verify that the sucrase activity negatively affects the leavening ability of baker's yeast strains under high-sucrose conditions. Based on a certain level of osmotolerance, sucrase level plays a significant role in the fermentation performance of baker's yeast, and appropriate sucrase activity is an important determinant for the leavening property of baker's yeast in sweet dough. Therefore, modification on sucrase activity is an effective method for improving the leavening properties of baker's yeast in sweet dough. This finding provides guidance for the breeding of industrial baker's yeast strains for sweet dough leavening. The transformants BS1 with deleted SUC2 genetic background provided decreased sucrase activity (a decrease of 39.3%) and exhibited enhanced leavening property (an increase of 12.4%). Such a strain could be useful for industrial applications.

MAL62 overexpression and NTH1 deletion enhance the freezing tolerance and fermentation capacity of the baker's yeast in lean dough.

BackgroundTrehalose is related to several types of stress responses, especially freezing response in baker’s yeast (Saccharomyces cerevisiae). It is desirable to manipulate trehalose-related genes to create yeast strains that better tolerate freezing-thaw stress with improved fermentation capacity, which are in high demand in the baking industry.ResultsThe strain overexpressing MAL62 gene showed increased trehalose content and cell viability after prefermention-freezing and long-term frozen. Deletion of NTH1 in combination of MAL62 overexpression further strengthens freezing tolerance and improves the leavening ability after freezing-thaw stress.ConclusionsThe mutants of the industrial baker’s yeast with enhanced freezing tolerance and leavening ability in lean dough were developed by genetic engineering. These strains had excellent potential industrial applications.

Near-freezing effects on the proteome of industrial yeast strains of Saccharomyces cerevisiae

At near-freezing temperatures (0–4°C), the growth of the yeast Saccharomyces cerevisiae stops or is severely limited, and viability decreases. Under these conditions, yeast cells trigger a biochemical response, in which trehalose and glycerol accumulate and protect them against severe cold and freeze injury. However, the mechanisms that allow yeast cells to sustain this response have been not clarified. The effects of severe cold on the proteome of S. cerevisiae have been not investigated and its importance in providing cell survival at near-freezing temperatures and upon freezing remains unknown. Here, we have compared the protein profile of two industrial baker’s yeast strains at 30°C and 4°C. Overall, a total of 16 proteins involved in energy-metabolism, translation and redox homeostasis were identified as showing increased abundance at 4°C. The predominant presence of glycolytic proteins among those upregulated at 4°C, likely represents a mechanism to maintain a constant supply of ATP for the synthesis of glycerol and other protective molecules. Accumulation of these molecules is by far the most important component in enhancing viability of baker’s yeast strains upon freezing. Overexpression of genes encoding certain proteins associated with translation or redox homeostasis provided specifically protection against extreme cold damage, underlying the importance of these functions in the near-freezing response.

Effects of SNF1 on Maltose Metabolism and Leavening Ability of Baker's Yeast in Lean Dough.

Maltose metabolism of baker's yeast (Saccharomyces cerevisiae) in lean dough is negatively influenced by glucose repression, thereby delaying the dough fermentation. To improve maltose metabolism and leavening ability, it is necessary to alleviate glucose repression. The Snf1 protein kinase is well known to be essential for the response to glucose repression and required for transcription of glucose-repressed genes including the maltose-utilization genes (MAL). In this study, the SNF1 overexpression and deletion industrial baker's yeast strains were constructed and characterized in terms of maltose utilization, growth and fermentation characteristics, mRNA levels of MAL genes (MAL62 encoding the maltase and MAL61 encoding the maltose permease) and maltase and maltose permease activities. Our results suggest that overexpression of SNF1 was effective to glucose derepression for enhancing MAL expression levels and enzymes (maltase and maltose permease) activities. These enhancements could result in an 18% increase in maltose metabolism of industrial baker's yeast in LSMLD medium (the low sugar model liquid dough fermentation medium) containing glucose and maltose and a 15% increase in leavening ability in lean dough. These findings provide a valuable insight of breeding industrial baker's yeast for rapid fermentation.

Effects of Over-liming on Wastewater Detoxification for Enhancement of Biogas Production

Detoxification by overliming was applied to enhance biogas production from an industrial baker’s yeast production wastewater. Various operational parameters affecting the detoxification, i.e., temperature, duration time, and pH, were studied for improvement of biogas production. Overliming was carried out at different pH levels (9 to 12) and temperatures (20 to 60°C) for different periods of time (0 to 12 h). Then, the detoxified wastewaters were subjected to anaerobic digestion. The maximum biogas production of 425 ml CH4/g COD, and the maximum yield of methane production of 3.06 l CH4/l wastewater were achieved under optimum conditions for overliming of wastewater.