Researchers in recent years have utilized a broad spectrum of treatment technologies in treating bakers' yeast production wastewater. This paper aims to review the treatment technologies for the wastewater, compare the process technologies, discuss recent innovations, and propose future perspectives in the research area. The review observed that nanofiltration was the most effective membrane process for the treatment of the effluent (at >95% pollutant rejection). Other separation processes like adsorption and distillation had technical challenges of desorption, a poor fit for high pollutant load and cost limitations. Chemical treatment processes have varying levels of success but they are expensive and produce toxic sludge. Sludge production would be a hurdle when product recovery and reuse are targeted. It is difficult to make an outright choice of the best process for treating the effluent because each has its merits and demerits and an appropriate choice can be made when all factors are duly considered. The process intensification of the industrial-scale production of the bakers' yeast process will be a very direct approach, where the process optimisation, zero effluent discharge, and enhanced recovery of value-added product from the waste streams are important approaches that need to be taken into account.
Glaciozyma antarctica PI12 is a psychrophilic yeast isolated from Antarctica. In this work, we describe the heterologous production, biochemical properties and in silico structure analysis of an arginase from this yeast (GaArg). GaArg is a metalloenzyme that catalyses the hydrolysis of L-arginine to L-ornithine and urea. The cDNA of GaArg was reversed transcribed, cloned, expressed and purified as a recombinant protein in Escherichia coli. The purified protein was active against L-arginine as its substrate in a reaction at 20 °C, pH 9. At 10-35 °C and pH 7-9, the catalytic activity of the protein was still present around 50%. Mn2+, Ni2+, Co2+ and K+ were able to enhance the enzyme activity more than two-fold, while GaArg is most sensitive to SDS, EDTA and DTT. The predicted structure model of GaArg showed a very similar overall fold with other known arginases. GaArg possesses predominantly smaller and uncharged amino acids, fewer salt bridges, hydrogen bonds and hydrophobic interactions compared to the other counterparts. GaArg is the first reported arginase that is cold-active, facilitated by unique structural characteristics for its adaptation of catalytic functions at low-temperature environments. The structure and function of cold-active GaArg provide insights into the potentiality of new applications in various biotechnology and pharmaceutical industries.
Simultaneous Saccharification and Fermentation (SSF) is a process where microbes have to first excrete extracellular enzymes to break polymeric substrates such as starch or cellulose into edible nutrients, followed by in situ conversion of those nutrients into more valuable metabolites via fermentation. As such, SSF is very attractive as a one-pot synthesis method of biological products. However, due to the co-existence of multiple biochemical steps, modeling SSF faces two major challenges. The first is to capture the successive chain-end and/or random scission of the polymeric substrates over time, which determines the rate of generation of various fermentable substrates. The second is to incorporate the response of microbes, including their preferential substrate utilization, to such a complex broth. Each of the above-mentioned challenges has manifested itself in many related areas, and has been competently but separately attacked with two diametrically different tools, i.e., the Population Balance Modeling (PBM) and the Cybernetic Modeling (CM), respectively. To date, they have yet to be applied in unison on SSF resulting in a general inadequacy or haphazard approaches to examine the dynamics and interactions of depolymerization and fermentation. To overcome this unsatisfactory state of affairs, here, the general linkage between PBM and CM is established to model SSF. A notable feature is the flexible linkage, which allows the individual PBM and CM models to be independently modified to the desired levels of detail. A more general treatment of the secretion of extracellular enzyme is also proposed in the CM model. Through a case study on the growth of a recombinant Saccharomyces cerevisiae capable of excreting a chain-end scission enzyme (glucoamylase) on starch, the interlinked model calibrated using data from the literature (Nakamura et al., Biotechnol. Bioeng. 53:21-25, 1997), captured features not attainable by existing approaches. In particular, the effect of various enzymatic actions on the temporal evolution of the polymer distribution and how the microbes respond to the diverse polymeric environment can be studied through this framework.
A divergent population of Saccharomyces cerevisiae has been identified in Malaysia by molecular and genetic analysis. It has also demonstrated that the yeast S. bayanus may be found in South America. Problems of the origin of S. cerevisiae are discussed.
In the field of nanotechnology, the use of various biological units instead of toxic chemicals for the reduction and stabilization of nanoparticles, has received extensive attention. Among the many possible bio resources, biologically active products from fungi and yeast represent excellent scaffolds for this purpose. Since fungi and yeast are very effective secretors of extracellular enzymes and number of species grow fast and therefore culturing and keeping them in the laboratory are very simple. They are able to produce metal nanoparticles and nanostructure via reducing enzyme intracellularly or extracellularly. The focus of this review is the application of fungi and yeast in the green synthesis of inorganic nanoparticles. Meanwhile the domain of biosynthesized nanoparticles is somewhat novel; the innovative uses in nano medicine in different areas including the delivery of drug, cancer therapy, antibacterial, biosensors, and MRI and medical imaging are reviewed. The proposed signaling pathways of nanoparticles induced apoptosis in cancerous cells and anti-angiogenesis effects also are reviewed. In this article, we provide a short summary of the present study universally on the utilization of eukaryotes like yeast and fungi in the biosynthesis of nanoparticles (NPs) and their uses.
Analyzing metabolic pathways in systems biology requires accurate kinetic parameters that represent the simulated in vivo processes. Simulation of the fermentation pathway in the Saccharomyces cerevisiae kinetic model help saves much time in the optimization process. Fitting the simulated model into the experimental data is categorized under the parameter estimation problem. Parameter estimation is conducted to obtain the optimal values for parameters related to the fermentation process. This step is essential because insufficient identification of model parameters can cause erroneous conclusions. The kinetic parameters cannot be measured directly. Therefore, they must be estimated from the experimental data either in vitro or in vivo. Parameter estimation is a challenging task in the biological process due to the complexity and nonlinearity of the model. Therefore, we propose the Artificial Bee Colony algorithm (ABC) to estimate the parameters in the fermentation pathway of S. cerevisiae to obtain more accurate values. A metabolite with a total of six parameters is involved in this article. The experimental results show that ABC outperforms other estimation algorithms and gives more accurate kinetic parameter values for the simulated model. Most of the estimated kinetic parameter values obtained from the proposed algorithm are the closest to the experimental data.
Ageing-related proteins play various roles such as regulating cellular ageing, countering oxidative stress, and modulating signal transduction pathways amongst many others. Hundreds of ageing-related proteins have been identified, however the functions of most of these ageing-related proteins are not known. Here, we report the identification of proteins that extended yeast chronological life span (CLS) from a screen of ageing-related proteins. Three of the CLS-extending proteins, Ptc4, Zwf1, and Sme1, contributed to an overall higher survival percentage and shorter doubling time of yeast growth compared to the control. The CLS-extending proteins contributed to thermal and oxidative stress responses differently, suggesting different mechanisms of actions. The overexpression of Ptc4 or Zwf1 also promoted rapid cell proliferation during yeast growth, suggesting their involvement in cell division or growth pathways.
Fermentation of sugar by Saccharomyces cerevisiae, for production of ethanol in an immobilized cell reactor (ICR) was successfully carried out to improve the performance of the fermentation process. The fermentation set-up was comprised of a column packed with beads of immobilized cells. The immobilization of S. cerevisiae was simply performed by the enriched cells cultured media harvested at exponential growth phase. The fixed cell loaded ICR was carried out at initial stage of operation and the cell was entrapped by calcium alginate. The production of ethanol was steady after 24 h of operation. The concentration of ethanol was affected by the media flow rates and residence time distribution from 2 to 7 h. In addition, batch fermentation was carried out with 50 g/l glucose concentration. Subsequently, the ethanol productions and the reactor productivities of batch fermentation and immobilized cells were compared. In batch fermentation, sugar consumption and ethanol production obtained were 99.6% and 12.5% v/v after 27 h while in the ICR, 88.2% and 16.7% v/v were obtained with 6 h retention time. Nearly 5% ethanol production was achieved with high glucose concentration (150 g/l) at 6 h retention time. A yield of 38% was obtained with 150 g/l glucose. The yield was improved approximately 27% on ICR and a 24 h fermentation time was reduced to 7 h. The cell growth rate was based on the Monod rate equation. The kinetic constants (K(s) and mu(m)) of batch fermentation were 2.3 g/l and 0.35 g/lh, respectively. The maximum yield of biomass on substrate (Y(X-S)) and the maximum yield of product on substrate (Y(P-S)) in batch fermentations were 50.8% and 31.2% respectively. Productivity of the ICR were 1.3, 2.3, and 2.8 g/lh for 25, 35, 50 g/l of glucose concentration, respectively. The productivity of ethanol in batch fermentation with 50 g/l glucose was calculated as 0.29 g/lh. Maximum production of ethanol in ICR when compared to batch reactor has shown to increase approximately 10-fold. The performance of the two reactors was compared and a respective rate model was proposed. The present research has shown that high sugar concentration (150 g/l) in the ICR column was successfully converted to ethanol. The achieved results in ICR with high substrate concentration are promising for scale up operation. The proposed model can be used to design a lager scale ICR column for production of high ethanol concentration.
There are multiple RNA degradation mechanisms in eukaryotes, key among these is mRNA decapping, which requires the Dcp1-Dcp2 complex. Decapping is involved in various processes including nonsense-mediated decay (NMD), a process by which aberrant transcripts with a premature termination codon are targeted for translational repression and rapid decay. NMD is ubiquitous throughout eukaryotes and the key factors involved are highly conserved, although many differences have evolved. We investigated the role of Aspergillus nidulans decapping factors in NMD and found that they are not required, unlike Saccharomyces cerevisiae. Intriguingly, we also observed that the disruption of one of the decapping factors, Dcp1, leads to an aberrant ribosome profile. Importantly this was not shared by mutations disrupting Dcp2, the catalytic component of the decapping complex. The aberrant profile is associated with the accumulation of a high proportion of 25S rRNA degradation intermediates. We identified the location of three rRNA cleavage sites and show that a mutation targeted to disrupt the catalytic domain of Dcp2 partially suppresses the aberrant profile of Δdcp1 strains. This suggests that in the absence of Dcp1, cleaved ribosomal components accumulate and Dcp2 may be directly involved in mediating these cleavage events. We discuss the implications of this.
Comparative genomic studies have reported widespread variation in levels of gene expression within and between species. Using these data to infer organism-level trait divergence has proven to be a key challenge in the field. We have used a wild Malaysian population of S. cerevisiae as a test bed in the search to predict and validate trait differences based on observations of regulatory variation. Malaysian yeast, when cultured in standard medium, activated regulatory programs that protect cells from the toxic effects of high iron. Malaysian yeast also showed a hyperactive regulatory response during culture in the presence of excess iron and had a unique growth defect in conditions of high iron. Molecular validation experiments pinpointed the iron metabolism factors AFT1, CCC1, and YAP5 as contributors to these molecular and cellular phenotypes; in genome-scale sequence analyses, a suite of iron toxicity response genes showed evidence for rapid protein evolution in Malaysian yeast. Our findings support a model in which iron metabolism has diverged in Malaysian yeast as a consequence of a change in selective pressure, with Malaysian alleles shifting the dynamic range of iron response to low-iron concentrations and weakening resistance to extreme iron toxicity. By dissecting the iron scarcity specialist behavior of Malaysian yeast, our work highlights the power of expression divergence as a signpost for biologically and evolutionarily relevant variation at the organismal level. Interpreting the phenotypic relevance of gene expression variation is one of the primary challenges of modern genomics.
Terpenes and terpenoids are among the key impact substances in the food and fragrance industries. Equipped with pharmacological properties and applications as ideal precursors for the biotechnological production of natural aroma chemicals, interests in these compounds have been escalating. Hence, the syntheses of new derivatives that can show improved properties are often called for. Stereoselective biotransformation offers several benefits to increase the rate of production, in terms of both the percentage yield and its enantiomeric excesses. Baker's yeast (Saccharomyces cerevisiae) is broadly used as a whole cell stereospecific reduction biocatalyst, due to its capability in reducing carbonyls and carbon-carbon double bonds, which also extends its functionality as a versatile biocatalyst in terpenoid biotransformation. This review provides some insights on the development and prospects in the reductive biotransformation of monoterpenoids and sesquiterpenoids using S. cerevisiae, with an overview of strategies to overcome the common challenges in large-scale implementation.
The evolutionarily conserved DNA polymerase delta (Polδ) plays several essential roles in eukaryotic DNA replication and repair, responsible for the synthesis of the lagging-strand, lower replicative mutagenesis via its proof-reading exonuclease activity and synthetizes both strands during break-induced replication. In Saccharomyces cerevisiae, the Polδ protein complex consists of three subunits encoded by the POL3, POL31 and POL32 genes. Surprisingly, in contrast to POL3 and POL31, the POL32 gene deletion was found to be viable but lethal in all other eukaryotes, raising the question to which extent the viability of the POL32 deletion in S. cerevisiae was species specific. To address this issue, we inactivated the POL32 gene in 10 evolutionary close or distant S. cerevisiae strains and found that POL32 was either essential (3 strains including SK1), non-essential (5 strains including the reference S288C strain) or confers a slow-growth phenotype (2 strains). Whole-genome sequencing of S288C/SK1 pol32∆ meiotic segregants identified the lethal/suppressor effect of the single Pol31-C43Y polymorphism. Consistently, the introduction of the Pol31-43C allele in the SK1 and West African (WA) pol32∆ mutants was sufficient to restore cell viability and wild-type growth upon introduction of two copies of POL31-43C in the SK1 haploid strain. Reciprocally, introduction of the SK1 POL31-43Y allele in the S288C pol32∆ mutant was lethal. Sequence analyses of the POL31 polymorphisms in the 1,011 yeasts genome dataset correlates with the strict occurrence of the POL31-43Y allele in the yeast African palm wine clade. Differently, the single Pol31-E400G polymorphism confers pol32∆ lethality in the Malaysian strain. In the yeast two-hybrid assay, we observed a weakened interaction between Pol3 and Pol31-43Y versus Pol31-43C suggesting an insufficient level of the Polδ holoenzyme stability/activity. Thus, the enigmatic non-essentiality of Pol32 in S. cerevisiae results from single Pol31 amino acid polymorphism and is clade rather than species specific.
Colloidal gas aphrons (CGAs) are highly stable, spherical, micrometer-sized bubbles encapsulated by surfactant multilayers. They have several intriguing properties, including: high stability, large interfacial area, and the ability to maintain the same charge as their parent molecules. The physical properties of CGAs make them ideal for biotechnological applications such as the recovery of a variety of: biomolecules, particularly proteins, yeast, enzymes, and microalgae. In this review, the bio-application of CGAs for the recovery of natural components is presented, as well as: experimental results, technical challenges, and critical research directions for the future. Experimental results from the literature showed that the recovery of biomolecules was mainly determined by electrostatic or hydrophobic interactions between polyphenols and proteins (lysozyme, β-casein, β-lactoglobulin, etc.), yeast, biological molecules (gallic acid and norbixin), and microalgae with CGAs. Knowledge transfer is essential for commercializing CGA-based bio-product recovery, which will be recognized as a viable technology in the future.
Red pitaya juice (RPJ) was subjected to UV-C irradiation and the potential of UV as a pasteurization tool for reducing microbial load in pitaya juice was evaluated. Effectiveness of the hurdle concept, i.e. addition of citric acid (CA) and dimethyl dicarbonate (DMDC) was also studied. Total plate counts (TPC) and yeast and mould counts (YMC) achieved 2.43 log₁₀ and 2.7 log₁₀ reductions respectively after exposure to UV irradiation. Addition of the CA (0.5 - 2.0%) and dimethyl dicarbonate (DMDC) (5 - 20 µL/100mL) to pitaya juice reduced the microbial loads, with 1.5% CA and 15 µL/100mL DMDC being the most effective concentrations. Addition of CA and DMDC into RPJ prior to UV treatment achieved significantly higher microbial reduction compared to UV alone, which were 4.12 log ₁₀ and 4.14 log₁₀ reductions for TPC and for YMC, respectively.
The proximate compositions, total dietary fibre (TDF) content, textural properties and sensory acceptability of yeast breads formulated with 0%, 2%, 4% and 6% of cornsilk powder (CSP) were studied. The protein, ash and TDF contents of yeast breads were increased in line with the CSP level added whereas moisture content was decreased. Yeast bread added with 6% CSP recorded the highest content of TDF (5.91%), protein (9.76%) and ash (1.03%) compared to other formulation of yeast breads containing lower percentage of CSP. Besides, texture profile analysis (TPA) reported that the firmness, gumminess and chewiness of yeast breads increased directly proportional to the level of CSP added mainly due to higher content of TDF and lower content of moisture. However, for the yeast bread added with 2%CSP, there were no significant differences compared with control yeast bread. Among all cornsilk-based yeast bread, formulation containing 2% CSP had the highest scores for all attributes including overall acceptance and there were no significant differences with control yeast bread. The present study indicated that the addition of 2% CSP could be an effective way to produce functional yeast bread without changing negatively its desirable textural and sensory acceptability.
This study is conducted to investigate the effect of different concentrations of betel leaves extract on color, pH and microbiological in homemade chili bo. The homemade chili bo with different concentrations (0 mg/ml, 0.75 mg/ml, 1.25 mg/ml and 1.75 mg/ml) of betel leaves extract were prepared for analysis. The results showed that the color of chili bo became darker as the concentration of betel leaves extract increased. The extract showed significant in the pH of chili bo after 7 days in which the highest concentration of extract showed the highest value of pH 4.31. The aerobic microbial count was decreased as the concentration of betel leaves extract increased in chili bo. After 7 days of storage, the highest concentration of betel leaves extract showed the highest percentage of reduction (6%), while the control sample showed 2.41% of aerobic reduction. The study also found that the extract contain lesser yeast and mold count (5.22 log CFU/ml) in homemade chili bo compared to the control sample (5.31 log CFU/ml) after 7 days. Betel leaves extract can be considered as natural food preservatives in chili bo to reduce the growth of spoilage microorganism and thus enhance the shelf life of chili bo.
A study was carried out to observe the fermentation process for noni (Morinda citrifolia L.) extract by Saccharomyces cerevisiae. The experiment was based on a central composite rotatable design (CCRD) employing 5 center points with augmented axial and factorial points resulting in 30 runs. The M. citrifolia extract was fermented with different combination of substrate concentration (40, 50, 60, 70 and 80%) (w/v), inoculum size (0, 1.5, 3, 4.5 and 6%) (v/v), temperature (30, 33.5, 37, 40.5 and 44oC) and fermentation time (0, 1.5, 3, 4.5 and 6 days). Five physico-chemical characteristics which include pH, titratable acidity, turbidity, total soluble solids and total polyphenol content were measured. Results showed that all the responses could be well represented using statistical models. For pH, only fermentation time was found to be not significant, while for titratable acidity and total polyphenol content, the effects of substrate concentration and fermentation time were significant. The effects of inoculum size and temperature level were found to be significant for turbidity. For total soluble solids, only the effect of substrate concentration and inoculum size were found to be significant.
Yeast growth and biomass production are greatly influenced by the length of the
incubation period during cultivation. Therefore, this study was conducted to
investigate the growth kinetics of five Lipomyces starkeyi strains as determined by
biomass production. The five L. starkeyi strains, namely L. starkeyi ATCC 12659, L.
starkeyi MV-1, L. starkeyi MV-4, L. starkeyi MV-5 and L. starkeyi MV-8, were inoculated
in sterilized Yeast Malt broth, and, incubated for 192 hr at ambient temperature.
Biomass yields were assessed and calculated gravimetrically every 24 hr. Results
indicated that the optimal biomass production of L. starkeyi ATCC 12659, L. starkeyi
MV-1, L. starkeyi MV-4, L. starkeyi MV-5 and L. starkeyi MV-8 were at 120, 168, 144,
168 and 120 hr, with the concentrations of 6.64, 6.43, 9.78, 11.23 and 8.56 g/L,
respectively. These results indicate that each L. starkeyi strain requires specific
incubation period for the optimum production of fungal biomass. Therefore, by
cultivating each L. starkeyi strain at the predetermined incubation period, biomass
yields could significantly be improved for further downstream applications such as
single cell protein and lipid production.
Kajian ini dijalankan untuk menentukan keupayaan yis (Saccharomyces cerevisiae) yang telah dipencil daripada buahbuahan tempatan iaitu duku langsat (Lansium domesticum), rambutan (Nephelium lappaceum), mangga Chokanan (Mangifera indica cv. Chokanan) dan rebung buluh minyak (Bambusa vulgaris) sebagai agen penaik roti berbanding roti kawalan penambahan yis komersial. Isi padu tertentu roti kawalan menunjukkan perbezaan bererti (p<0.05) dengan semua roti kajian. Roti yang difermentasi oleh yis rebung buluh menunjukkan isi padu tertentu yang paling tinggi, diikuti oleh roti yang difermentasi oleh yis mangga dan yis duku langsat dan ketiga-tiga yis tersebut juga mempunyai isi padu tertentu yang lebih tinggi secara bererti (p<0.05) dengan roti kawalan. Roti yang difermentasi oleh yis rebung buluh dan yis mangga mempunyai tekstur yang lebih lembut berbanding roti kawalan. Peningkatan dalam isi padu tertentu roti boleh meningkatkan kelembutan tekstur roti. Peratus kandungan kelembapan kulit dan isi roti yang difermentasi oleh yis rebung buluh adalah paling tinggi manakala roti yang difermentasi oleh yis rambutan menunjukkan peratus kandungan kelembapan yang paling rendah. Peningkatan peratus kandungan kelembapan juga boleh meningkatkan kelembutan tesktur roti dan sebaliknya. Daripada segi warna kulit, hanya kecerahan warna kulit (L*) roti yang difermentasi oleh yis rebung buluh tidak menunjukkan perbezaan bererti (p>0.05) dengan warna kulit roti kawalan. Kecerahan warna isi (L*) roti kawalan pula menunjukkan perbezaan bererti (p<0.05) dengan semua isi roti kajian. Selain itu, didapati semakin kecil dan padat liang udara, semakin putih warna isi roti. Keseragaman taburan liang udara juga menghasilkan isi roti yang lebih putih. Secara keseluruhan, kesemua yis yang dipencil berpotensi untuk dijadikan sebagai agen penaik. Yis rebung buluh dan yis mangga dapat menghasilkan kualiti roti putih yang lebih baik daripada yis komersial.
Kajian keberkesanan sifat antimikrob ekstrak kacangma dijalankan menggunakan ujian resapan cakera dan ujian perencatan langsung. Hasil menunjukkan ekstrak etanol dengan kepekatan 50 dan 100 mg/mL merencat Staphylococcus aureus. Bagi ekstrak air, kepekatan 10, 25, 50 dan 100 mg/mL merencat Aspergillus niger, 25, 50 dan 100 mg/mL dapat merencat Saccharomyces cerevisae dan kepekatan 100 mg/mL dapat merencat Staphylococcus aureus. Perlakuan suhu yang berbeza ke atas ekstrak dalam ujian perencatan langsung tidak menunjukkan sebarang perbezaan ke atas perencatan mikroorganisma yang dikaji.