Yeast Candida Lipolytica Research Articles

The effect of local soybean fermentation duration using yeast on total plate count, temperature, ph, and water content

This research aims to determine the best use of yeast and fermentation time for Total Plate Count (TPC), temperature, pH, and water content in fermented local soybeans. This research used a Completely Randomized Design with a nested pattern with two factors. Factor 1 uses yeast Saccharomyces cerevisiae and Candida lipolytica, and factor 2 is the fermentation time of 2, 4, and 8 days and repeated four times, so there are 24 treatment units. The data was analyzed using analysis of variance and continued with the Least Significant Difference test. The research results showed that TPC produced an average value of 17.75×109 CFU/mL 37.75×109 CFU/mL, pH 4.69 6.51, temperature 26.775 30.925 °C, water content 32.51% 48.74%. The yeast type S. cerevisiae, with a fermentation time of 4 days, produces the best water content, while C. lipolytica with a fermentation time of 4 days produces the best TPC and temperature. Using C. lipolytica with a fermentation time of 8 days produces the best pH.

Alkaline hydrolysis of olive fruits wastes for the production of single cell protein by Candida lipolytica

This study aims to investigate the probable production of single-cell protein (SCP) through an identified yeast Candida lipolytica 10 DSM 70561, SM-0301 through the hydro-alkaline decomposition of the olive residuals by the recycling of the olive fruit residues and its use as a carbonic source for yeast growth and to provide the ideal circumstance to gain highest quantity of Carbohydrate of the hydro-alkali product. Alternation of the physical and chemical factors of the nutritional media in order to obtain the highest rate of production. The ideal state of this hydro-decomposition was as follows: 0.8 M of sodium hydroxide, temperature at 100 °C and duration time of 60 min. This context provided the highest sugar production, i.e. up to 12.5%. The ideal growth of the yeast and production of SCP was after 4 days of incubation with a rotation speed of 150 RPM at 30 °C and initial pH value of 5.5. Moreover, the medium containing 0.4% peptone as a source was superiors than other used peptone concentration with respect to yeast growth (17.55 g/l) and SCP production (11.24 g/l).

Photophysical and antibacterial activity of light-activated quaternary eosin Y

AbstractThe functional characteristics of a new eosin dye with biocidal quaternary ammonium group (E) were studied in aqueous solution and in organic solvents of different polarity. The spectral properties depend on the nature and polarity of the respective solvents. The antimicrobial activity of compound E has been tested in vitro against Gram-negative bacteria (Escherichia coli, Acinetobacter johnsoni and Pseudomonas aeruginosa), Gram-positive bacteria (Sarcina lutea and Bacillus cereus) and the antifungal activity was tested against the yeasts Candida lipolytica in solution and after treated on cotton fabric. Broth dilution test has been used for quantitative evaluation of the antimicrobial activity of compound E against the model strains. The ability of compound E to inhibit the growth of model Gram-negative P. aeruginosa strain was assessed after 16 h of incubation in presence and absence of light. These experiments were conducted in planktonic format in solution and on cotton fabric. The results suggest that the new compound is effective in treating the relevant pathogens with better results being obtained by irradiation with light. In this case the quaternary ammonium group promotes the binding of eosin Y moiety to the bacterial cell wall thus accelerating bacterial photo inactivation.

PH sensor potential and antimicrobial activity of a new PPA dendrimer modified with benzanthrone fluorophores in solution and on viscose fabric

The paper reports on the influence of pH on the absorption and fluorescence characteristics of a new poly(propylene amine) dendrimer (BD) from second generation modified with eight fluorescent benzanthrone chromophore units. Spectral characteristics have been determined using various percentages of the DMF-water mixture. The antimicrobial activity of the newly synthesized dendrimer BD has been determined in vitro against Gram-positive bacteria Bacillus cereus, Gram-negative bacteria Pseudomonas aeruginosa, and the yeasts Candida lipolytica used as test cultures. Viewing production of antibacterial textile, the antimicrobial activity of the dendrimer deposited on the surface of viscous fabric has also been investigated.

Yeasts and bacterial biosurfactants as demulsifiers for petroleum derivative in seawater emulsions

Oil sludge or waste generated in transport, storage or refining forms highly stable mixtures due to the presence and additives with surfactant properties and water forming complex emulsions. Thus, demulsification is necessary to separate this residual oil from the aqueous phase for oil processing and water treatment/disposal. Most used chemical demulsifiers, although effective, are environmental contaminants and do not meet the desired levels of biodegradation. We investigated the application of microbial biosurfactants as potential natural demulsifiers of petroleum derivatives in water emulsions. Biosurfactants crude extracts, produced by yeasts (Candida guilliermondii, Candida lipolytica and Candida sphaerica) and bacteria (Pseudomonas aeruginosa, Pseudomonas cepacia and Bacillus sp.) grown in industrial residues, were tested for demulsification capacity in their crude and pure forms. The best results obtained were for bacterial biosurfactants, which were able to recover about 65% of the seawater emulsified with motor oil compared to 35–40% only for yeasts products. Biosurfactants were also tested with oil-in-water (O/W) and water-in-oil (W/O) kerosene model emulsions. No relationship between interfacial tension, cell hydrophobicity and demulsification ratios was observed with all the biosurfactants tested. Microscopic illustrations of the emulsions in the presence of the biosurfactants showed the aspects of the emulsion and demulsification process. The results obtained demonstrate the potential of these agents as demulsifiers in marine environments.

Waste Soybean Oil and Corn Steep Liquor as Economic Substrates for Bioemulsifier and Biodiesel Production by Candida lipolytica UCP 0998.

Almost all oleaginous microorganisms are available for biodiesel production, and for the mechanism of oil accumulation, which is what makes a microbial approach economically competitive. This study investigated the potential that the yeast Candida lipolytica UCP0988, in an anamorphous state, has to produce simultaneously a bioemulsifier and to accumulate lipids using inexpensive and alternative substrates. Cultivation was carried out using waste soybean oil and corn steep liquor in accordance with 22 experimental designs with 1% inoculums (107 cells/mL). The bioemulsifier was produced in the cell-free metabolic liquid in the late exponential phase (96 h), at Assay 4 (corn steep liquor 5% and waste soybean oil 8%), with 6.704 UEA, IE24 of 96.66%, and showed an anionic profile. The emulsion formed consisted of compact small and stable droplets (size 0.2–5 µm), stable at all temperatures, at pH 2 and 4, and 2% salinity, and showed an ability to remove 93.74% of diesel oil from sand. The displacement oil (ODA) showed 45.34 cm2 of dispersion (central point of the factorial design). The biomass obtained from Assay 4 was able to accumulate lipids of 0.425 g/g biomass (corresponding to 42.5%), which consisted of Palmitic acid (28.4%), Stearic acid (7.7%), Oleic acid (42.8%), Linoleic acid (19.0%), and γ-Linolenic acid (2.1%). The results showed the ability of C. lipopytica to produce both bioemulsifier and biodiesel using the metabolic conversion of waste soybean oil and corn steep liquor, which are economic renewable sources.

Synthesis and spectral characterization of a new PPA dendrimer modified with 4-bromo-1,8-naphthalimide and in vitro antimicrobial activity of its Cu(II) and Zn(II) metal complexes

This article describes the peripheral modification of a polypropylene amine dendrimer of second generation with eight 4-bromo-1,8-naphthalimide units and its copper and zinc complexes. The new dendrimer and its metal complexes have been characterized by electronic (UV–vis and fluorescence) and vibrational (IR and Raman) spectral methods in view of their antibacterial and antifungal activity. The results have shown that the two metal ions form complexes with the dendrimer ligand. In vitro antimicrobial screening of the newly synthesized metallodendrimers showed promising antimicrobial activities against some pathogenic Gram-positive and Gram-negative bacteria and the yeasts Candida lipolytica. The results suggest that the new metal complexes could find application in designing new antimicrobial preparations to control the spread of infections caused especially by Acinetobacter, Micrococcus and Candida species.

Synthesis and evaluation of biosurfactant produced by Candida lipolytica using animal fat and corn steep liquor

Abstract In the present study, low-cost media based on animal fat and corn steep liquor combined with glucose, yeast extract, urea and other inorganic nitrogen sources were evaluated for the production of biosurfactants by the yeast Candida lipolytica UCP0988 in batch shake flasks. At the end of the 6-day fermentation period, the medium containing only animal fat (5%) and corn steep liquor (2.5%) yielded the maximal reduction in surface tension (from 50 to 28 mN/m). The properties of the biosurfactant separated by different organic solvent extractions were investigated and the critical micelle concentration was determined. Preliminary chemical characterisation revealed the anionic nature of the biosurfactant. Compositional analysis of the biosurfactant was carried out using thin layer chromatography. The biosurfactant produced by the isolate was characterised as a glycolipid. Emulsification activity and surface tension stability of the biosurfactant produced using different hydrocarbons and vegetable oils and the effect of pH, temperature and the addition of salt were also studied. The cell-free broth (crude biosurfactant) was effective at recovering up to 70% of the residual oil from oil-saturated sand samples and also effective at oil displacement (54%). The crude biosurfactant from C. lipolytica and aqueous solutions of the isolated biosurfactant at 0.04%, 0.08% and 0.16% were effective in recovering up to 100% of the motor oil from the walls of the beakers. These properties indicate the potential of a cheaply produced glycolipid for application in the oil industry.

FTIR, XRD, and DSC analysis of the rosemary extract effect on polyethylene structure and biodegradability

Differential scanning calorimetry (DSC) technique enabled the study of the effect of the rosemary (Rosmarinus officinalis) extract on polyethylene foils structure and biodegradability. The polyethylene used for food packaging contains synthetic antioxidants which are often able to migrate and to contaminate the food. Natural extracts with antioxidant properties may be an interesting alternative for the polyethylene fabrication. The influence of the rosemary present in different concentrations in non-irradiated and irradiated polyethylene samples was studied. The biodegradability studies were carried out by incubating the modified polyethylene with the yeast Candida lipolytica. The changes of physical properties and morphological aspects before and after fungal attack were studied by Fourier Transform Infrared Spectroscopy, X-ray diffraction, and DSC. With some exceptions due to the nonuniform character of the investigated samples, the melting peak temperature and crystallization degree were found to decrease with increasing of rosemary content and irradiation. Synergic effects of rosemary and irradiation on the film biodegradability were directly confirmed by optical microscopy.

Antimicrobial and antioxidant activities of lignin from residue of corn stover to ethanol production

To improve the economic viability of the biofuel production from biomass resource, a value-added lignin byproduct from this process is increasingly important. Antioxidant and antimicrobial activities of lignin extracted from residue of corn stover to ethanol production were investigated. The lignin extracts exhibited strong antioxidant activities in hydrophilic oxygen radical absorbance capacity (ORAC) assay and Folin–Ciocalteu test. The extracts also exhibited antimicrobial activities against Gram-positive bacteria ( Listeria monocytogenes and Staphylococcus aureus) and yeast ( Candida lipolytica), but not Gram-negative bacteria ( Escherichia coli O157:H7 and Salmonella Enteritidis) or bacteriophage MS2. Different extraction conditions (temperature and residue/solvent ratio) affected the antioxidant and antimicrobial activities of lignin extracts. Generally, the bioactivities of lignin extracts were consistent with FTIR analysis results. Lignin byproducts showed the potential for their antioxidant and antimicrobial application.

Antimicrobial and anti-adhesive potential of a biosurfactant Rufisan produced by Candida lipolytica UCP 0988

In the last years, researches developed with biosurfactants for application in the medical area have been revealing the promising biological activities of these biomolecules. In this work the antimicrobial and anti-adhesive properties of a biosurfactant Rufisan isolated from the yeast Candida lipolytica UCP 0988, growth in a medium supplemented with ground nut refinery residue was determined against several microorganisms. The biosurfactant was able to reduce the water surface tension from 70 to 25.3mN/m and showed a critical micelle concentration (CMC) of 0.03%. The biosurfactant was isolated after 72h of fermentation and was tested in concentrations varying from 0.75 to 12mg/l. The highest antimicrobial activities were observed against Streptococcus agalactiae, Streptococcus mutans, Streptococcus mutans NS, Streptococcus mutans HG, Streptococcus sanguis 12, Streptococcus oralis J22 at a concentration superior to the biosurfactant critical micelle concentration. Moreover, the biosurfactant showed anti-adhesive activity against most of the microorganisms tested. As far as we know, this is the first compilation of data on antimicrobial and anti-adhesive activities of a biosurfactant obtained from a Candida strain against such a broad group of microorganisms. The results obtained in this work showed that the biosurfactant from C. lipolytica is a potential antimicrobial and/or anti-adhesive agent for several biomedical applications.

Combined pH and high hydrostatic pressure effects on Lactococcus starter cultures and Candida spoilage yeasts in a fermented milk test system during cold storage

The combined effects of high pressure processing (HPP) and pH on the glycolytic and proteolytic activities of Lactococcus lactis subsp. lactis, a commonly used cheese starter culture and the outgrowth of spoilage yeasts of Candida species were investigated in a fermented milk test system. To prepare the test system, L. lactis subsp. lactis C10 was grown in UHT skim milk to a final pH of 4.30 and then additional samples for treatment were prepared by dilution of fermented milk with UHT skim milk to pH levels of 5.20 and 6.50. These milk samples (pH 4.30, 5.20 and 6.50) with or without an added mixture of two yeast cultures, Candida zeylanoides and Candida lipolytica (10 5 CFU mL −1 of each species), were treated at 300 and 600 MPa (≤20 °C, 5 min) and stored at 4 °C for up to 8 weeks. Continuing acidification by starter cultures, as monitored during storage, was substantially reduced in the milk pressurised at pH 5.20 where the initial titratable acidity (TA) of 0.40% increased by only 0.05% (600 MPa) and 0.10% (300 MPa) at week 8, compared to an increase of 0.30% in untreated controls. No substantial differences were observed in pH or TA between pressure-treated and untreated milk samples at pH 4.30 or 6.50. The rate of proteolysis in milk samples at pH values of 5.20 and 6.50 during storage was significantly reduced by treatment at 600 MPa. Treatment at 600 MPa also reduced the viable counts of both Candida yeast species to below the detection limit (1 CFU mL −1) at all pH levels for the entire storage period. However, samples treated at 300 MPa showed recovery of C. lipolytica from week 3 onwards, reaching 10 6–10 7 CFU mL −1 by week 8. In contrast, C. zeylanoides did not show any recovery in any of the pressure-treated samples during storage.

TEA EXTRACTS AS POSSIBLE NATURAL FOOD PRESERVATIVE FOR ORGANIC FOOD

The inhibitory effect of hot water extracts of green and black tea on some food spoilage and poisoning microorganisms was assayed using the in vitro zone inhibition technique. The antimicrobial effect of tea extracts was evaluated on various representatives of different microbial groups i.e. filamentous fungi (Aspergillus niger, Aspergillus oryzae, Fusarium moniliforme, Trichoderma viride & Trichoderma reesei), yeasts (Candida lipolytica, Candida utilis, Geotricum candidum & Saccharomyces cerevisiae), gram positive bacteria (Bacillus cereus & Staphylococcus aureus) and gram negative bacteria (Erwinia carotovora and Escherichia coli). Generally, extracts of the black tea showed stronger antifungal action on the different tested fungi than that of the green tea. Also, the highest concentrations of both tested tea extracts gave the strongest antifungal action. However, Staphylococcus aureus was the only gram positive bacteria which were inhibited by the higher tested concentrations of black tea, while green tea gave no inhibition effect. Whereas, the different tested concentrations of black tea inhibited the tested gram negative bacteria i.e. Erwinia carotovora and E. coli while green tea extracts at 3 and 4 % concentration inhibited only E. coli. Also, C. lipolytica was the only tested yeast which expressed growth inhibition at the different tested concentrations of both green and black tea extracts. It could be concluded that tea extracts could be used as natural food preservative in the processed organic food and/or beverage, which might be subjected to fungal spoilage.

Bioemulsifier production in batch culture using glucose as carbon source by Candida lipolytica.

The yeast Candida lipolytica IA 1055 produced an inducible extracellular emulsification activity while utilizing glucose at different concentrations as carbon source during batch fermentation at 27 degrees C. In all glucose concentrations studied, maximum production of emulsification activity was detected in the stationary phase of growth, after pH reached minimal values. The bioemulsifier isolated was a complex biopolymer constituting proteins, carbohydrates, and lipids. The results obtained in this work show that the biosynthesis of a bioemulsifier is not simply a prerequisite for the degradation of extracellular hydrocarbon.

Effect of Growth Phase on the Content and Composition of Ceramides of the Hydrocarbon-assimilating Yeast Candida lipolytica

Candida lipolytica yeast was grown batchwise on n-hexadecane as the carbon and energy source. Ceramides were quantitatively isolated from total lipids of exponential and stationary phase cells by a combination of column chromatography and preparative high-performance thin-layer chromatography. After acid methanolysis their composition was analyzed by gas-liquid chromatography. The ceramide content of the exponential phase cells was two times higher than the one of the stationary phase cells. The composition of long-chain base moiety of ceramides did not change significantly during the growth. In both growth phases 19-phytosphingosine was the major long-chain base. However, the fatty acid composition of ceramides changed greatly during the growth. In the exponential growth phase, ceramides contained predominantly fatty acids greater than 20 carbon atoms, while fatty acids shorter than 20 atoms predominated in ceramides of the stationary phase, 16:0 being the main one. In the exponential growth phase fatty acid moiety of ceramides was characterized by unusually high degree of unsaturation and relatively high proportion of odd-numbered fatty acids. However, the proportion of both, unsaturated and odd-numbered fatty acid decreased significantly in ceramides of the stationary phase. The unexpected finding was the absence of fatty acid hydroxylation of ceramides in the exponential phase cells and unusually low degree of hydroxylation in the stationary phase.

Enzymes for the resolution of α-tertiary-substituted carboxylic acid esters

Aromatic α-amino-α-methyl acids and α-hydrazino-α-methyl acids are known aromatic amino acid decarboxylase inhibitors. Specific derivatives such as 2-amino-2-methyl-3-(3,4-dihydroxyphenyl)propanoate, Aldomet®, and 2-hydrazino-2-methyl-3-(3,4-dihydroxypheny Lodosyn®, have been developed as therapeutic agents to treat hypertension and Parkinson's disease, respectively. We recently reported a method for the kinetic resolution of the racemic esters of such compounds using a crude preparation of a novel enzyme catalyst from the yeast Candida lipolytica (Yee, C.; Blythe, T. A.; McNabb, T. J.; Walts, A. E. J. Org. Chem. 1992, 57, 3525-3527). Here we report the purification and initial characterization of the active enzyme component, an enzyme given the name Candida lipolytica ester hydrolase (CLEH). CLEH was purified to > 95% homogeneity by chromatography on Matrex TM Blue B resin. The enzyme was found to be a glycoprotcin with M r = 80,000–300,000. In addition to esterolytic activity, the enzyme was found to catalyze the hydrolysis of amides, anilides and peptides. Sequence analysis of internal peptides of CLEH revealed striking homology to a number of enzymes belonging to the group of serine carboxypeptidases (E.G. 3.4.16.1). One peptide aligned with the canonical serine carboxypeptidase active site sequence, GESYAG. Based on the structural relationship of CLEH to serine carboxypeptidases, three representative serine carboxypeptidases were evaluated for their utility in resolving racemic α-tertiary ester substrates and compared with the activity of CLEH. All enzymes revealed similarly high activity and enantioselectivity towards the α-hydrazino-α-methyl ester precursor of the Parkinson-drug Carbidopa. However, differences in enantioselectivity were observed with other α-tertiary-substituted ester substrates. Serine carboxypeptidase-catalyzed ester resolutions thus offer a new route to many sterically hindered homochiral α-amino, α-hydrazino and α-hydroxy carboxylic acids.

Lipid hydrogenation induces elevated 18:1-CoA desaturase activity in Candida lipolytica microsomes

Microsomal membranes prepared from the mesophilic yeast Candida lipolytica grown at 10 degrees C were hydrogenated by the homogeneous Pd-catalyst, palladium di (sodium alizarine sulfonate) (Pd(QS)2). After hydrogenation to various levels, the microsomes were washed free of the Pd-complex and transferred to a reaction mixture (containing NADH, MgCl2, ATP, CoA and [14C]18:1-CoA) for assay of 18:1-CoA desaturase activity. Microviscosity alterations were also followed by measuring changes in DPH fluorescence polarization. Rapid catalytic hydrogenation of unsaturated fatty acids of the lipids occurred within 20-120 s, resulting in large increases in 16:0, 18:0 and 18:1 acids and decreases in 18:2 acid. In the range 7-20% 18:0 content, a pronounced increase in desaturase activity was observed, with a maximum of greater than 2-fold at a 18:0 content of 12%, followed by a decrease to the initial activity at 33% 18:0 content. These changes were well-correlated with changes in microviscosity, maximal desaturase activity occurring in the DPH fluorescence anisotropy range of 0.23-0.24; above and below this range, desaturase activities were close to the initial control values. It is suggested that the hydrogenation-induced increase in the formation of 18:2 from 18:1-CoA (proceeding partly through direct desaturation of PC) may be due to changes in conformation of the membrane-bound desaturase enzyme complex as a result of controlled rigidification of the surrounding lipids. The operation of such a self-regulating control mechanism would be consistent with a previously proposed model for microsomal desaturase action.

Characterization of A cAMP-independent Ca 2+-inhibited protamine kinase from [formula omitted

A cAMP-independent protamine kinase has been purified from extracts of the yeast Candida lipolytica by ion-exchange and affinity chromatography. Two subunits with apparent M r's of 52,000 and 36,000 were resolved by SDS-PAGE. The purified kinase exhibited about 20% activity with casein and histone Type VII-S as substrates relative to protamine. The enzyme was inactive against other protein substrates tested, and was essentially insensitive to AMP, cAMP, cGMP up to 0.2 mM, the polyamines spermine and spermidine up to 1 mM, N-ethylmaleimide (5 mM), 2-mercaptoethanol (20 mM), or dithiothreitol (2 mM), and several cations like Zn 2+, N 1+, or Co 2+ at 0.1 mM each. Ca 2+ at 3 mM inhibited protamine kinase activity by 50%, which was reversed by EGTA.

Purification and Characterization of Liposan, a Bioemulsifier from Candida lipolytica

The inducible water-soluble bioemulsifier liposan (M. C. Cirigliano and G. M. Carman, Appl. Environ. Microbiol. 48:747-750, 1984) was purified from the yeast Candida lipolytica. The purification procedure included repeated solvent extractions of a concentrated culture filtrate and Affi-Gel concanavalin A affinity chromatography. The procedure yielded a preparation containing a major band (M(r) = 27,600) which stained for protein and carbohydrate upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Liposan is composed of approximately 83% carbohydrate and 17% protein. Acid and enzymatic digestions of the emulsifier revealed that the carbohydrate portion is a heteropolysaccharide consisting of glucose, galactose, galactosamine, and galacturonic acid. Liposan effected and stabilized oil-in-water emulsions with a variety of commercial vegetable oils. Emulsification and stabilization properties of liposan were compared to those of a number of commercial emulsifiers and stabilizers.

Recovery of heat‐stressed yeasts in media containing plant oleoresins

Conner, D.E. & Beuchat, L.R. 1985. Recovery of heat‐stressed yeasts in media containing plant oleoresins. Journal of Applied Bacteriology59, 49–55.Oleoresins from seven plants (allspice, cinnamon, clove, garlic, onion, oregano, and thyme) were tested for their effects on eight genera of sublethally heated food‐spoilage and industrially important yeasts (Candida lipolytica, Debaryomyces han‐senii, Hansenula anomala, Kloeckera apiculata, Lodderomyces elongisporus, Rhodotorula rubra, Saccharomyces cerevisiae, and Torulopsis glabrata). All heat‐stressed yeasts had increased sensitivity to oleoresins in recovery media. Heated cells were most sensitive to cinnamon oleoresin, a lethal effect being observed to concentrations as low as 5 μg/ml. Results indicate that all eight yeasts underwent metabolic and/or structural injury as a result of exposure to elevated temperature. Whilst changes in sensitivity to oleoresins were not the same for all yeasts, it is concluded that the oleoresins may have a pronounced synergistic or additive effect on thermal inactivation of yeasts in foods. Furthermore, carryover of oleoresins in low dilutions of food to enumeration media could adversely influence the recovery of heat‐stressed cells, thus resulting in apparent low populations.