Despite the advantages of continuous fermentation whereby ethanol is selectively removed from the fermenting broth to reduce the end-product inhibition, this process can concentrate minor secondary products to the point where they become toxic to the yeast. This study aims to develop a new mathematical model do describe the inhibitory effect of byproducts on alcoholic fermentation including glycerol, lactic acid, acetic acid, and succinic acid, which were reported as major byproducts during batch alcoholic fermentation. The accumulation of these byproducts during the different stages of batch fermentation has been quantified. The yields of total byproducts, glycerol, acetic acid, and succinic acid per gram of glucose were 0.0442, 0.023, 0.0155, and 0.0054, respectively. It was found that the concentration of these byproducts linearly increases with the increase in glucose concentration in the range of 25-250 g/L. The results have also showed that byproduct concentration has a significant inhibitory effect on specific growth coefficient (μ) whereas no effect was observed on the half-velocity constant (Ks). A new mathematical model of alcoholic fermentation was developed considering the byproduct inhibitory effect, which showed a good performance and more accuracy compared to the classical Monod model.
Results of a biodiversity study of Antarctic microfungi from ornithogenic soils are presented in this paper. A wide range of soil habitats within and adjacent to active and abandoned penguin rookeries were sampled in order to examine relationships between environmental factors and the biodiversity of soil microfungi. Soil samples were collected from two contrasting Antarctic locations: (1) Beaufort Island (Ross Sea, Continental Antarctica), which is largely ice- and snow-covered, isolated, difficult to access and infrequently visited, and (2) Barrientos Island (maritime Antarctica) which is mostly ice-free during summer and is often visited by scientists and tourists. Soil sampling at Beaufort and Barrientos Islands was completed during the austral summer seasons of 2004/05 and 2006/07, respectively. Warcup’s soil method was used for fungi cultivation. A total of 27 fungal taxa were isolated from the two study sites, consisting of 11 ascomycetes, 13 hyphomycetes
and three yeasts. Only three taxa — Geomyces sp., a pink and a white yeast — occurred on both sites. The isolated fungi were classified according to their thermal characteristics in culture, with seven psychrophilic, 10 psychrotrophic and 10 mesophilic fungi being isolated. Thelebolus microspores, Thelebolus sp., Geomyces sp. and Antarctomyces sp., were the most frequently isolated fungi. A total of 10 taxa were isolated from the 20 soil samples from Beaufort Island, consisting of five psychrophilic, four psychrotrophic and one mesophilic fungi. Thelebolus microsporus, Thelebolus sp., Asco BI8 and Phoma sp. were the most frequently obtained fungi
(20%–27% of isolates). A total of 22 fungal taxa were isolated from 23 soil samples from Barrientos Island, consisting of four psychrophilic, six psychrotrophic and 12 mesophilic fungi. Geomyces sp. and Antarctomyces sp. were the most frequently isolated taxa. Thus, the fungal diversity of Beaufort Island was dominated by Ascomycetes while that of Barrientos Island was dominated by hyphomycetes.
Lipid biosynthesis and fatty acids composition of oleaginous zygomycetes, namely Cunninghamella bainieri 2A1, cultured in media with excess or limited nitrogen were quantitatively determined at different times of culture growth. Accumulation of lipids occurred even when the activity of NAD(+)-ICDH (β-Nicotinamide adenine dinucleotide-isocitrate dehydrogenase) was still detectable in both media. In C. bainieri 2A1, under nitrogen limitation, the ratio of lipids was around 35%, whereas in nitrogen excess medium (feeding media supplemented with ammonium tartarate), the lipid ratio decreased. The amount of this decrease depended on the level of ammonium tartarate in the media. The main findings in this paper were that C. bainieri 2A1 has the ability to accumulate lipid although nitrogen concentration detected inside the media and that NAD-ICDH was active in all culture periods. These results proved that the strain C. bainieri 2A1 has an alternative behavior in lipid biosynthesis that differs from yeast. According to the old hypotheses, yeasts could not accumulate lipid more than 10% when nitrogen was detected inside the media. Nitrogen-limited and excess media both contained the same fatty acids (palmitic acid, stearic acid, olic acid, linoleic acid and γ-linolenic acid), but at different concentrations. The C:N ratio was also studied and showed no effects on total lipid accumulation, but a significant effect on γ-linolenic acid concentration.
Automated methods for reconstructing biological networks are becoming increasingly important in computational systems biology. Public databases containing information on biological processes for hundreds of organisms are assisting in the inference of such networks. This paper proposes a multiobjective genetic algorithm method to reconstruct networks related to metabolism and protein interaction. Such a method utilizes structural properties of scale-free networks and known biological information about individual genes and proteins to reconstruct metabolic networks represented as enzyme graph and protein interaction networks. We test our method on four commonly-used protein networks in yeast. Two are networks related to the metabolism of the yeast: KEGG and BioCyc. The other two datasets are networks from protein-protein interaction: Krogan and BioGrid. Experimental results show that the proposed method is capable of reconstructing biological networks by combining different omics data and structural characteristics of scale-free networks. However, the proposed method to reconstruct the network is time-consuming because several evaluations must be performed. We parallelized this method on GPU to overcome this limitation by parallelizing the objective functions of the presented method. The parallel method shows a significant reduction in the execution time over the GPU card which yields a 492-fold speedup.
The interest in utilizing algae for wastewater treatment has been increased due to many advantages. Algae-wastewater treatment system offers a cost-efficient and environmentally friendly alternative to conventional treatment processes such as electrocoagulation and flocculation. In this biosystem, algae can assimilate nutrients in the wastewater for their growth and simultaneously capture the carbon dioxide from the atmosphere during photosynthesis resulting in a decrease in the greenhouse gaseousness. Furthermore, the algal biomass obtained from the treatment process could be further converted to produce high value-added products. However, the recovery of free suspended algae from the treated effluent is one of the most important challenges during the treatment process as the current methods such as centrifugation and filtration are faced with the high cost. Immobilization of algae is a suitable approach to overcome the harvesting issue. However, there are some drawbacks with the common immobilization carriers such as alginate and polyacrylamide related to low stability and toxicity, respectively. Hence, it is necessary to apply a new carrier without the mentioned problems. One of the carriers that can be a suitable candidate for the immobilization is zeolite. To date, various types of zeolite have been used for the immobilization of cells of bacteria and yeast. If there is any possibility to apply them for the immobilization of algae, it needs to be considered in further studies. This article reviews cell immobilization technique, biomass immobilization onto zeolites, and algal immobilization with their applications. Furthermore, the potential application of zeolite as an ideal carrier for algal immobilization has been discussed.
Antifreeze proteins (AFPs) are specific proteins, glycopeptides, and peptides made by different organisms to allow cells to survive in sub-zero conditions. AFPs function by reducing the water's freezing point and avoiding ice crystals' growth in the frozen stage. Their capability in modifying ice growth leads to the stabilization of ice crystals within a given temperature range and the inhibition of ice recrystallization that decreases the drip loss during thawing. This review presents the potential applications of AFPs from different sources and types. AFPs can be found in diverse sources such as fish, yeast, plants, bacteria, and insects. Various sources reveal different α-helices and β-sheets structures. Recently, analysis of AFPs has been conducted through bioinformatics tools to analyze their functions within proper time. AFPs can be used widely in various aspects of application and have significant industrial functions, encompassing the enhancement of foods' freezing and liquefying properties, protection of frost plants, enhancement of ice cream's texture, cryosurgery, and cryopreservation of cells and tissues. In conclusion, these applications and physical properties of AFPs can be further explored to meet other industrial players. Designing the peptide-based AFP can also be done to subsequently improve its function.
Iron is an essential micronutrient for most living beings since it participates as a redox active cofactor in many biological processes including cellular respiration, lipid biosynthesis, DNA replication and repair, and ribosome biogenesis and recycling. However, when present in excess, iron can participate in Fenton reactions and generate reactive oxygen species that damage cells at the level of proteins, lipids and nucleic acids. Organisms have developed different molecular strategies to protect themselves against the harmful effects of high concentrations of iron. In the case of fungi and plants, detoxification mainly occurs by importing cytosolic iron into the vacuole through the Ccc1/VIT1 iron transporter. New sequenced genomes and bioinformatic tools are facilitating the functional characterization, evolution and ecological relevance of metabolic pathways and homeostatic networks across the Tree of Life. Sequence analysis shows that Ccc1/VIT1 homologs are widely distributed among organisms with the exception of animals. The recent elucidation of the crystal structure of a Ccc1/VIT1 plant ortholog has enabled the identification of both conserved and species-specific motifs required for its metal transport mechanism. Moreover, recent studies in the yeast Saccharomyces cerevisiae have also revealed that multiple transcription factors including Yap5 and Msn2/Msn4 contribute to the expression of CCC1 in high-iron conditions. Interestingly, Malaysian S. cerevisiae strains express a partially functional Ccc1 protein that renders them sensitive to iron. Different regulatory mechanisms have been described for non-Saccharomycetaceae Ccc1 homologs. The characterization of Ccc1/VIT1 proteins is of high interest in the development of biofortified crops and the protection against microbial-derived diseases.
A number of living creatures in the Antarctic region have developed characteristic adaptation of cold weather by producing antifreeze proteins (AFP). Antifreeze peptide (Afp1m) fragment have been designed in the sequence of strings from native proteins. The objectives of this study were to assess the properties of Afp1m to cryopreserve skin graft at the temperature of -10 °C and -20 °C and to assess sub-zero injuries in Afp1m cryopreserved skin graft using light microscopic techniques. In the present study, a process was developed to cryopreserve Sprague-Dawley (SD) rat skin grafts with antifreeze peptide, Afp1m, α-helix peptide fragment derived from Glaciozyma antractica yeast. Its viability assessed by different microscopic techniques. This study also described the damages caused by subzero temperatures (-10 and -20 °C) on tissue cryopreserved in different concentrations of Afp1m (0.5, 1, 2, 5 and 10 mg/mL) for 72 h. Histological scores of epidermis, dermis and hypodermis of cryopreserved skin grafts showed highly significant difference (p
Foodborne illness is recognized as an emerging infectious disease. The incidence of foodborne
infections is common and the majority cases are undiagnosed or unreported. Apart from some
diarrhea or minor gastrointestinal problem, some foodborne pathogenic microbes may cause
death, particularly to those people with weakened immune system. In this study, we have
developed a new fermented papaya beverage using symbiotic culture of yeast and acetic acid
bacteria under controlled biofermentation process. An in-vitro assessment of fermented papaya
beverage against few foodborne pathogenic microorganism was conducted to determine
its minimum bactericidal concentration (MBC>99). Three types of foodborne pathogen:
Escherichia coli O157, Salmonella enterica serovar Typhimurium ATCC 53648, Salmonella
enterica serovar Enteritidis (isolated from infectious chicken) were selected. From minimum
bactericidal concentration (MBC>99) assay, both fermented papaya pulp and leaves beverages
have shown 100% killing rate against three selected foodborne pathogenic microbes. Inversely,
non-fermented papaya pulp and leaves beverages indicated no inhibition at all. In fact, further
dilution of fermented papaya pulp and leaves beverages demonstrated different degree of
MBC>99 and brix value, but the pH value remained less than 3.5. These findings indicated
the combination of soluble solid compounds presents in both fermented papaya beverage and
product acidity play an important role in the inhibition of pathogenic microorganisms. The
preliminary promising results of this work have shown that the great potential of fermented
papaya beverages as a preventive measure to reduce the incidence of foodborne illness.
Widespread food poisoning due to microbial contamination has been a major concern for the food industry, consumers and governing authorities. This study is designed to determine the levels of fungal contamination in edible bird nests (EBNs) using culture and molecular techniques. Raw EBNs were collected from five house farms, and commercial EBNs were purchased from five Chinese traditional medicine shops (companies A-E) in Peninsular Malaysia. The fungal contents in the raw and commercial EBNs, and boiled and unboiled EBNs were determined. Culturable fungi were isolated and identified. In this study, the use of these methods revealed that all EBNs had fungal colony-forming units (CFUs) that exceeded the limit set by Standards and Industrial Research Institute of Malaysia (SIRIM) for yeast and moulds in EBNs. There was a significant difference (p < 0.05) in the number of types of fungi isolated from raw and commercial EBNs, but no significant difference in the reduction of the number of types of fungi after boiling the EBNs (p > 0.05). The types of fungi isolated from the unboiled raw EBNs were mainly soil, plant and environmental fungi, while the types of fungi isolated from the boiled raw EBNs, unboiled and boiled commercial EBNs were mainly environmental fungi. Aspergillus sp., Candida sp., Cladosporium sp., Neurospora sp. and Penicillum sp. were the most common fungi isolated from the unboiled and boiled raw and commercial EBNs. Some of these fungi are mycotoxin producers and cause opportunistic infections in humans. Further studies to determine the mycotoxin levels and methods to prevent or remove these contaminations from EBNs for safe consumption are necessary. The establishment and implementation of stringent regulations for the standards of EBNs should be regularly updated and monitored to improve the quality of the EBNs and consumer safety.
As fermentation could reduce the negative effects of bran on final cereal products, the utilization of whole-cereal flour is recommended, such as brown rice flour as a functional food ingredient. Therefore, this study aimed to investigate the effect of fermented brown rice flour on white rice flour, white rice batter and its steamed bread qualities. Brown rice batter was fermented using commercial baker's yeast (Eagle brand) according to the optimum conditions for moderate acidity (pH 5.5) to obtain fermented brown rice flour (FBRF). The FBRF was added to white rice flour at 0%, 10%, 20%, 30%, 40% and 50% levels to prepare steamed rice bread. Based on the sensory evaluation test, steamed rice bread containing 40% FBRF had the highest overall acceptability score. Thus, pasting properties of the composite rice flour, rheological properties of its batter, volume and texture properties of its steamed bread were determined. The results showed that peak viscosity of the rice flour containing 40% FBRF was significantly increased, whereas its breakdown, final viscosity and setback significantly decreased. Viscous, elastic and complex moduli of the batter having 40% FBRF were also significantly reduced. However, volume, specific volume, chewiness, resilience and cohesiveness of its steamed bread were significantly increased, while hardness and springiness significantly reduced in comparison to the control. These results established the effectiveness of yeast fermentation in reducing the detrimental effects of bran on the sensory properties of steamed white rice bread and encourage the usage of brown rice flour to enhance the quality of rice products.
Overexpression of MSN2, which is the transcription factor gene in response to stress, is well-known to increase the tolerance of the yeast Saccharomyces cerevisiae cells to a wide variety of environmental stresses. Recent studies have found that the Msn2 is a feasible potential mediator of proline homeostasis in yeast. This result is based on the finding that overexpression of the MSN2 gene exacerbates the cytotoxicity of yeast to various amino acid analogs whose uptake is increased by the active amino acid permeases localized on the plasma membrane as a result of a dysfunctional deubiquitination process. Increased understanding of the cellular responses induced by the Msn2-mediated proline incorporation will provide better comprehension of how cells respond to and counteract to different kinds of stimuli and will also contribute to the breeding of industrial yeast strains with increased productivity.
Plants biosynthesize a great diversity of biologically active small molecules of interest for fragrances, flavors, and pharmaceuticals. Among specialized metabolites, terpenoids represent the greatest molecular diversity. Many terpenoids are very complex, and total chemical synthesis often requires many steps and difficult chemical reactions, resulting in a low final yield or incorrect stereochemistry. Several drug candidates with terpene skeletons are difficult to obtain by chemical synthesis due to their large number of chiral centers. Thus, biological production remains the preferred method for industrial production for many of these compounds. However, because these chemicals are often found in low abundance in the native plant, or are produced in plants which are difficult to cultivate, there is great interest in engineering increased production or expression of the biosynthetic pathways in heterologous hosts. Although there are many examples of successful engineering of microbes such as yeast or bacteria to produce these compounds, this often requires extensive changes to the host organism's metabolism. Optimization of plant gene expression, post-translational protein modifications, subcellular localization, and other factors often present challenges. To address the future demand for natural products used as drugs, new platforms are being established that are better suited for heterologous production of plant metabolites. Specifically, direct metabolic engineering of plants can provide effective heterologous expression for production of valuable plant-derived natural products. In this review, our primary focus is on small terpenoids and we discuss the benefits of plant expression platforms and provide several successful examples of stable production of small terpenoids in plants.
Fennel (Foeniculum vulgare Mill.) and coriander (Coriandrum sativum L.) are known to possess good antimicrobial properties. In the present work, spice-infused frozen parathas were formulated to investigate the effect of fennel and coriander on microbial (aerobic mesophilic bacteria, yeast and mould, and Bacillus cereus) reduction and sensory acceptability of frozen paratha throughout the storage at -18°C. The present work was also aimed at determining the relationship between spice concentrations and storage durations on microbiological quality of the samples. Fennel and coriander seed powder were used at concentrations of 2, 4 and 6% of wheat flour (w/w). The microbiological analysis was performed by total plate count, yeast and mould count, and Bacillus cereus count after 9, 12 and 15 weeks of storage. Sensory evaluation was conducted using hedonic scales at the end of storage durations. Results showed that spice infusion in frozen paratha significantly delayed the growth of aerobic mesophilic bacteria, yeasts and moulds, and Bacillus cereus during storage. The lowest log count was demonstrated by coriander at 6% in total plate count (3.85, 3.90 and 3.91 log10 CFU/g), and yeast and mould count (2.54, 2.59 and 2.60 log10 CFU/g) after 9, 12 and 15 weeks, respectively. Bacillus cereus was not detected throughout the storage durations. Fennel exhibited minimum activity against Bacillus cereus with no significant difference on log count reduction when compared with control. Coriander showed the highest decrease in both total plate count and Bacillus cereus count during the storage duration. Sensory evaluation result indicated that control sample exhibited the highest preference over all attributes when compared with fennel and coriander. Coriander-infused paratha was slightly darker in colour due to high concentration of 6%. Fennel yielded the lowest score in terms of taste among all samples. Fennel and coriander showed no significant difference for sensory acceptability. Overall, all frozen parathas were in good quality after 15 weeks of frozen storage. It can thus be concluded that fennel and coriander can be used as potential natural preservatives to inhibit the growth of microorganisms in paratha during frozen storage. Nevertheless, the optimum spice concentration should be determined to minimise the effects on the sensory attributes.
Supercritical carbon dioxide (SC-CO2 ) is a non-thermal technique implemented by food, pharmaceutical, and similar industries with the aim of inhibiting the microorganisms and apply effective sterilisation. Presently, limited number of studies has reported the application of SC-CO2 on fresh chicken meat. The present work therefore aimed to reveal the microbial and physicochemical quality of the SC-CO2 -treated fresh chicken meat. The fresh chicken meat was subjected to the SC-CO2 at 14 MPa and 45°C for 40 min and was stored at 4°C for 0, 3, and 7 days. The obtained results indicated that the treatment with SC-CO2 significantly decreased the total plate count and, yeast and mould count from log10 5.90 to 2.00 CFU/g and from log10 5.02 to 2.00 CFU/g at day 7 of storage, respectively. The values of pH, cooking loss, and water holding capacity were not affected by the treatment. The results revealed that the SC-CO2 -treated samples displayed harder texture, higher lightness and yellowness, and lower redness. In addition, lipid peroxidation of SC-CO2 and control samples resulted in values of 1.9 and 0.5 MDA/mg of meat at day 7 of storage time and did not significantly change in the rest of the evaluation days. In summary, the application of SC-CO2 was capable of enhancing the microbial quality and certain physicochemical attributes. However, alteration of certain parameters of SC-CO2 might enhance the overall meat quality.
Green coffee beans are stored for a certain period and under certain conditions until they are finally utilized. The storage period may depend on customer demand while the storage conditions depend on where the coffee beans are stored. Thus, this research emphasizes the physicochemical changes that occur in Liberica coffee beans during storage under the Malaysian
climate (average temperature and relative humidity of 29.33ºC and 71.75% respectively). The changes in the physico-chemical (coffee size, mass, densities, colour, proximate analysis, sucrose, chlorogenic acid content) and microbiological (yeast and mould count) properties were evaluated during eight months of storage. After the storage, the physical properties of the coffee changed as the coffee beans expanded in size, reduced in mass and density and became brighter in colour. Changes in the chemical properties were also detected where the moisture decreased and the ash content increased. In addition, the sucrose level was found to decrease with a corresponding increase in chlorogenic acid. During storage, the counts of yeast and mould were reduced. Model equations describing the changes in the properties were developed. The overall conclusion was that the coffee beans reduced in quality during storage.
Aflatoxins are carcinogenic, mutagenic and teratogenic fungal toxins predominantly produced by Aspergillus flavus (A. flavus) and Aspergillus parasiticus (A. parasiticus). Members of the Aspergillus family are wound-invading pathogens that can infect pistachio trees and nuts. The pistachio nut is a favorite tree nut worldwide, and more than half of the world’s pistachio production is from Iran. Pistachio nuts can easily be infected with Aspergillus spp. due to early splitting or due to animal, insect or physical damage. Any established infection of Aspergillus under high relative humidity and temperature results in the production and rapid accumulation of aflatoxins in pistachio nuts. It is impractical to remove aflatoxins from pistachio nuts after they are produced. Some microorganisms (such as saprophytic yeasts) have been reported to have an antagonistic effect against Aspergillus spp. This study aimed to isolate saprophytic yeasts from pistachio fruits and leaves and investigate their biocontrol activities against a toxigenic strain of Aspergillus flavus (A. flavus). Saprophytic yeasts were identified based on their morphological properties and biochemical tests. In total, 24 yeast isolates were obtained from pistachio fruits and leaves, and their antagonistic effect on A. flavus (PTCC 5006) was investigated. Five saprophytic yeast isolates, displaying the highest biocontrol activities against A. flavus (PTCC 5006), were identified as Pseudozyma fusiformata, Cryptococcus albidus, Rhodotorula fragaria, Cryptococcus hungaricus and Rhodotorula hinula. The biocontrol activities of these yeast isolates were evaluated by their inhibitory effects on sporulation, colony expansion, biomass production and prevention of aflatoxin B1 (AFB1) production. Pseudozyma fusiformata was the most effective yeast isolate in terms of spore reduction (84.6%) and inhibition of AFB1 production (89.1%). Cryptococcus albidus produced the maximum reduction in fungal dry weight (77.9%). Based on these results, isolated saprophytic yeasts from pistachio fruits and leaves can be used as effective biocontrol agents against the growth of Aspergillus and aflatoxin production.
Mid-exponential phase Saccharomyces rouxii YSa40 cells subsequently stressed at low aw/pH in the 0.64 aw/pH 3.5 glycerol/CPB system became injured. Such injury was detected by the loss of ability of the
stressed population to form colonies on secondary-stress plating medium (glycerol/BM agar at 0.94 aw
/pH 3.5 (lactic acid)) while colony forming ability on secondary non-stress plating medium (sugars/BM agar at 0.94 aw/pH 3.5 (lactic) was unaffected. The injury was shown to be due to sensitivity to glycerol/lactic acid. Results of the present study will be useful for achieving complete decontamination of ‘Intermediate Moisture Foods’ against xerotolerant molds and yeast.
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.
In this study polymerase chain reaction (PCR) was used to identify yeast in domestic ragi obtained
from two local markets in Sarawak and Pahang. These ragi are normally used as a dry starter in food fermentation (tapai) for Pahang (ST2) and Sarawak (ST3) and tuak (ST1) which is an alcoholic drink in Sarawak. Universal primer, NL1 and NL4 were used as a primer in this study to amplify D1/D2 fragment. Based on the result from the sequencing and after the BLAST search of the nucleotide sequences, the strain was confirmed as Candida glabrata (FN424108.) partial 26S rRNA gene, strain IMUFRJ 51955 for ST1, Saccharomyces cerevisiae(EU285514.1) isolate 35 26S ribosomal RNA gene, partial sequence for ST2 sample and Candida glabrata (FN393990.1) partial 26S rRNA gene, strain MUCL 51244 for ST3. All these strains were found in domestic ragi used for food fermentation.