This study was undertaken to optimize skim milk and yeast extract concentration as a cultivation medium for optimal Bifidobacteria pseudocatenulatum G4 (G4) biomass and β -galactosidase production as well as lactose and free amino nitrogen (FAN) balance after cultivation period. Optimization process in this study involved four steps: screening for significant factors using 2(3) full factorial design, steepest ascent, optimization using FCCD-RSM, and verification. From screening steps, skim milk and yeast extract showed significant influence on the biomass production and, based on the steepest ascent step, middle points of skim milk (6% wt/vol) and yeast extract (1.89% wt/vol) were obtained. A polynomial regression model in FCCD-RSM revealed that both factors were found significant and the strongest influence was given by skim milk concentration. Optimum concentrations of skim milk and yeast extract for maximum biomass G4 and β -galactosidase production meanwhile low in lactose and FAN balance after cultivation period were 5.89% (wt/vol) and 2.31% (wt/vol), respectively. The validation experiments showed that the predicted and experimental values are not significantly different, indicating that the FCCD-RSM model developed is sufficient to describe the cultivation process of G4 using skim-milk-based medium with the addition of yeast extract.
Vaginal discharge is a common complaint of women attending gynaecological clinics. The purpose of this study was to compare the occurrence of commonly implicated microorganisms in vaginal discharge amongst women with or without the complaint, attending a gynaecological and family planning clinic. The association of Gardnerella vaginalis with bacterial vaginosis was also studied. It was found that there were no significant differences between the cases and controls in the isolation rate of Gardnerella vaginalis, Torulopsis glabrata, Ureaplasma urealyticum, Mycoplasma ssp and Group B streptococcus (p greater than 0.05). Only the isolation rate of Candida albicans was significantly higher in the cases than controls (p less than 0.01). However, there was a significant association of G. vaginalis with bacterial vaginosis.
The yield and properties of cellulose produced from bacterial fermentation of black tea broth (known as Kombucha) were investigated in this study. The tea broth was fermented naturally over a period of up to 8 days in the presence of sucrose. Tea broth with a sucrose concentration of 90 g/l produced highest yield of bacterial cellulose (66.9%). The thickness and yield of bacterial cellulose increased with fermentation time. The bacterial cellulose production increased correspondingly with increased surface area:depth ratio. Changes in pH were related to the symbiotic metabolic activities of yeasts and acetic acid bacteria, and the counts of both of these in the tea broths were relatively higher than those in the cellulose layer. Findings from this study suggest that the yield of cellulose depends on many factors that need to be optimized to achieve maximum yield.
Ethanolic extract of Cassia alata leaves was investigated for its antimicrobial activities on several microorganisms including bacteria, yeast, dermatophytic fungi and non-dermatophytic fungi. In vitro, the extract exhibited high activity against various species of dermatophytic fungi but low activity against non-dermatophytic fungi. However, bacterial and yeast species showed resistance against in vitro treatment with the extract. The minimum inhibitory concentration (MIC) values of the extract revealed that Trichophyton mentagorphytes var. interdigitale, Trichophyton mentagrophytes var. mentagorophytes, Trichophyton rubrum and Microsporum gypseum had the MIC of 125 mg/ml, whereas Microsporum canis had the MIC of 62.5 mg/ml. The inhibition can be observed on the macroconidia of Microsporum gypseum which resulted in structural degeneration beyond repair. The mechanism of inhibition can be related to the cell leakage as observed by irregular, wrinkle shape and loss in rigidity of the macroconidia.
Bioprotein is one of the useful products obtained from biotechnology invention. It is a promising replacement for the commercial fish feed supplement. In this study, the enrichment of the bioprotein content after solid state fermentation using palm kernel cake and seaweed by the white rot fungus: Phanerochaete chrysoporium and yeast: Candida utilis was carried out. The growth media components were selected from 11 types of media using Plackett-Burman design (hereinafter PBD) and were optimized by one-factor-at-a-time (OFAT) method with bioprotein concentration (mg/g) as the response. From the screening result using PBD, three media components, namely K2HPO4, CuSO4.5H2O and MnSO4.H2O were selected for further optimization using OFAT method because of their positive contributions to the response. The final results showed that 5.0 g/L K2HPO4, 3.0 g/L CuSO4.5H2O and 0.1 g/L MnSO4.H2O were there to be the optimum media constituents with 9.0 g/L, MgSO4.7H2O, 0.1 g/L, CaCl2.H2O, 3.0 g/L FeSO4.7H2O and 3.0 g/L peptone as fixed compositions. At this optimum concentration, the protein increment of 11% was observed as compared to the results determined in the screening using PBD. The study revealed the benefits of using mixed cultures in improving the protein concentrations which can be used as nutritious fish feed.
A broad range of cell lines with characteristic features are used as bio-factories to produce recombinant proteins for basic research and therapeutic purposes. Genetic engineering strategies have been used to manipulate the genome of mammalian cells, insects, and yeasts for heterologous expression. One reason is that the glycosylation pattern of the expression hosts differs somehow from mammalian cells, which may cause immunogenic reactions upon administration in humans. CRISPR-Cas9 is a simple, efficient, and versatile genome engineering tool that can be programmed to precisely make double-stranded breaks at the desired loci. Compared to the classical genome editing methods, a CRISPR-Cas9 system is an ideal tool, providing the opportunity to integrate or delete genes from the target organisms. Besides broadened applications, limited studies have used CRISPR-Cas9 for editing the endogenous pathways in expression systems for biopharmaceutical applications. In the present review, we discuss the use of CRISPR-Cas9 in expression systems to improve host cell lines, increase product yield, and humanize glycosylation pathways by targeting intrinsic genes.
Condensin is a central regulator of mitotic genome structure with mutants showing poorly condensed chromosomes and profound segregation defects. Here, we identify NCT, a complex comprising the Nrc1 BET-family tandem bromodomain protein (SPAC631.02), casein kinase II (CKII), and several TAFs, as a regulator of condensin function. We show that NCT and condensin bind similar genomic regions but only briefly colocalize during the periods of chromosome condensation and decondensation. This pattern of NCT binding at the core centromere, the region of maximal condensin enrichment, tracks the abundance of acetylated histone H4, as regulated by the Hat1-Mis16 acetyltransferase complex and recognized by the first Nrc1 bromodomain. Strikingly, mutants in NCT or Hat1-Mis16 restore the formation of segregation-competent chromosomes in cells containing defective condensin. These results are consistent with a model where NCT targets CKII to chromatin in a cell-cycle-directed manner in order to modulate the activity of condensin during chromosome condensation and decondensation.
The extract of the Psophocarpus tetragonolobus pods has been tested for antimicrobial activity in a disk diffusion assay on eight human pathogenic bacteria and two human pathogenic yeasts. The extracts of P. tetragonolobus possessed antimicrobial activity against all tested strains. The ethanolic extract of P. tetragonolobus pods was further tested for in vivo brine shrimp lethality test and in vitro sheep erythrocyte cytotoxic assay. The brine shrimp lethality test exhibited no significant toxicity (LC(50)=1.88 mg/ml) against Artemia salina, whereas sheep erythrocyte test showed significant toxicity. The reason for haemolysis of erythrocyte was discussed. The P. tetragonolobus extract with high LC(50) value signified that this plant is not toxic to human. This result also suggested that the ethanolic extract of P. tetragonolobus pods is potential source for novel antimicrobial compounds.
Onychomycosis is the infection of nail apparatus by dermatophytes, yeasts or non-dermatophyte moulds and is responsible for 50% of all nail disorders. A five year retrospective study was conducted at Universiti Kebangsaan Malaysia to identify the common pathogens responsible for onychomycosis and to describe the epidemiology of the affected patients. A total of 278 abnormal nails were cultured, out of which 231 were positive for fungus. Females constituted 50.2% (n=116) while males 49.8% (n=115). The majority (51.9%, n=120) were between ages 50-69 years. The Malay ethnic group was most commonly affected (44.2%, n=102) followed by Chinese (33.8%, n=78), Indians (18.2%, n=42) and other ethnic groups (3.8%, n=9). The most common fungal element isolated was non-dermatophyte moulds (45.4%, n=105) followed by yeast (34.6%, n=80) and dermatophytes (1.3%, n=3). Aspergillus spp. was the commonest (59.8%,n=81) non-dermatophyte mould, while Candida spp. was the commonest yeast (74.3%, n=89) isolated. In this study, non-dermatophyte moulds are the most common microorganisms implicated to cause onychomysosis. Treatment for non-dermatophyte mould is challenging as the current available antifungal agents are more effective against dermatophytes and yeasts.
The biodiversity and the killer activity of yeasts isolated from various types of fermented food in Malaysia were investigated in this study. Of 252 yeasts isolated from 48 fermented food samples in this study, 19 yeast species were identified based on sequence analysis of the ITS1-5.8S-ITS2 partial fragments of the yeasts. A total of 29 (11.5%) of the yeast isolates demonstrated killer activity to at least one Candida species tested in this study; including 22 isolates of Trichosporon asahii, 4 isolates of Pichia anomala, and one isolate each of Pichia norvegensis, Pichia fermentans and Issatchenkia orientalis, respectively. The presence of killer yeasts reflects antagonism that occurs during microbial interaction in the fermented food, whereby certain yeasts produce killer toxins and possibly other toxic substances in competition for limited nutrients and space. The anti-Candida activity demonstrated by killer yeasts in this study should be further explored for development of alternative therapy against candidiasis.
The aim of this study was to investigate the presence of multidrug resistant yeasts in the faeces of synanthropic wild birds from the Bangsar suburb of Kuala Lumpur.
Cell contact formation, which is the process by which cells are brought into close proximity is an important biotechnological process in cell and molecular biology. Such manipulation is achieved by various means, among which dielectrophoresis (DEP) is widely used due to its simplicity. Here, we show the advantages in the judicious choice of the DEP microelectrode configuration in terms of limiting undesirable effects of dielectric heating on the cells, which could lead to their inactivation or death, as well as the possibility for cell clustering, which is particularly advantageous over the linear cell chain arrangement typically achieved to date with DEP. This study comprises of experimental work as well as mathematical modeling using COMSOL. In particular, we establish the parameters in a capillary-based microfluidic system giving rise to these optimum cell-cell contact configurations, together with the possibility for facilitating other cell manipulations such as spinning and rotation, thus providing useful protocols for application into microfluidic bioparticle manipulation systems for diagnostics, therapeutics or for furthering research in cellular bioelectricity and intercellular interactions.
Mixotrophic metabolism was evaluated as an option to augment the growth and lipid production of marine microalga Tetraselmis sp. FTC 209. In this study, a five-level three-factor central composite design (CCD) was implemented in order to enrich the W-30 algal growth medium. Response surface methodology (RSM) was employed to model the effect of three medium variables, that is, glucose (organic C source), NaNO3 (primary N source), and yeast extract (supplementary N, amino acids, and vitamins) on biomass concentration, X(max), and lipid yield, P(max)/X(max). RSM capability was also weighed against an artificial neural network (ANN) approach for predicting a composition that would result in maximum lipid productivity, Pr(lipid). A quadratic regression from RSM and a Levenberg-Marquardt trained ANN network composed of 10 hidden neurons eventually produced comparable results, albeit ANN formulation was observed to yield higher values of response outputs. Finalized glucose (24.05 g/L), NaNO3 (4.70 g/L), and yeast extract (0.93 g/L) concentration, affected an increase of X(max) to 12.38 g/L and lipid a accumulation of 195.77 mg/g dcw. This contributed to a lipid productivity of 173.11 mg/L per day in the course of two-week cultivation.
A total of 2,153 high vaginal swabs were processed for the presence of yeasts. The specimens were obtained from pregnant and non-pregnant Malaysian women with and without vaginitis. The yeast species most commonly isolated were Candida albicans, C. glabrata, C. famata and C. parapsilosis. C. albicans was isolated from 27% of pregnant women with vaginitis, 14% of pregnant women with no overt vaginitis, 15% of non-pregnant women with vaginitis, and 3% of non-pregnant women with no vaginitis. The significant difference of the isolation rates from women with and without vaginitis indicates that C. albicans is likely to be a vaginal pathogen.
The yeast species composition of 12 cocoa bean fermentations carried out in Brazil, Ecuador, Ivory Coast and Malaysia was investigated culture-independently. Denaturing gradient gel electrophoresis of 26S rRNA gene fragments, obtained through polymerase chain reaction with universal eukaryotic primers, was carried out with two different commercial apparatus (the DCode and CBS systems). In general, this molecular method allowed a rapid monitoring of the yeast species prevailing during fermentation. Under similar and optimal denaturing gradient gel electrophoresis conditions, the CBS system allowed a better separated band pattern than the DCode system and an unambiguous detection of the prevailing species present in the fermentation samples. The most frequent yeast species were Hanseniaspora sp., followed by Pichia kudriavzevii and Saccharomyces cerevisiae, independent of the origin of the cocoa. This indicates a restricted yeast species composition of the cocoa bean fermentation process. Exceptionally, the Ivorian cocoa bean box fermentation samples showed a wider yeast species composition, with Hyphopichia burtonii and Meyerozyma caribbica among the main representatives. Yeasts were not detected in the samples when the temperature inside the fermenting cocoa pulp-bean mass reached values higher than 45 °C or under early acetic acid production conditions.
The preparation and observations of spheroplast W303 cells are described with Environmental Scanning Electron Microscope (ESEM). The spheroplasting conversion was successfully confirmed qualitatively, by the evaluation of the morphological change between the normal W303 cells and the spheroplast W303 cells, and quantitatively, by determining the spheroplast conversion percentage based on the OD800 absorbance data. From the optical microscope observations as expected, the normal cells had an oval shape whereas spheroplast cells resemble a spherical shape. This was also confirmed under four different mediums, that is, yeast peptone-dextrose (YPD), sterile water, sorbitol-EDTA-sodium citrate buffer (SCE), and sorbitol-Tris-Hcl-CaCl2 (CaS). It was also observed that the SCE and CaS mediums had a higher number of spheroplast cells as compared to the YPD and sterile water mediums. The OD800 absorbance data also showed that the whole W303 cells were fully converted to the spheroplast cells after about 15 minutes. The observations of the normal and the spheroplast W303 cells were then performed under an environmental scanning electron microscope (ESEM). The normal cells showed a smooth cell surface whereas the spheroplast cells had a bleb-like surface after the loss of its integrity when removing the cell wall.
Consecutive chloroform, ethanol, and ethyl acetate partitions of extracts from winged bean [Psophocarpus tetragonolobus (L.) DC] root, stem, leaf, and pod extracts were tested for their antimicrobial activity against 19 microbial species, including 11 bacterial pathogens, four yeasts, and four molds using the disk diffusion assay technique. The pod extract was found to be most effective against all of the tested organisms, followed by the stem, root, and leaf extracts, and the ethanol fraction showed the most significant (p < 0.05) antimicrobial activity against all of the tests among three soluble fractions of extract, followed by the ethyl acetate and chloroform fractions. The minimum inhibitory concentrations (MICs) of extracts determined by the broth dilution method ranged from 1.25 to 10.0 mg/mL. The MIC of ethanol fraction of pod extracts was the lowest by comparison with the other two extracts. The MIC for fungi was at or below 2.5 mg/mL and for bacteria was at or above 2.5 mg/mL.
Matched MeSH terms: Yeasts/drug effects*; Yeasts/growth & development