Candida Bombicola Research Articles

PSVIII-33 Therapeutic effects of sophorolipid on intestinal restoration in HT-29 cell and early-weaned rats

Abstract Sophorolipid (SPL) is a glycolipid bio-surfactant produced by Candida bombicola and displays antimicrobial activity and stimulates collagenesis with less toxic and more biodegradable than other surfactants. As its effect has not been demonstrated in weaning animal models, we conducted two experiments to investigate the effect of SPL on intestinal restoration. First, a wound healing assay using HT-29 cells was conducted to quantify migration capacity. Cells were seeded into 6-well plate and scratched. After 24- and 48-hour incubation, SPL increased migration of cells in a dose-dependent manner (P < 0.05). It seemed that SPL had restorative effects on intestinal wounds. Afterwards, an in vivo experiment was conducted using thirty 2-week-old early-weaned rats, which were randomly allotted into three treatments (CON; NIH-31 diet, BAM; 32 ppm of bambermycin and SPL; 10 ppm of SPL). During 16 days of trial, rats fed ad libitum and body weights were measured every four days. Rat were sacrificed at days 8 and 16 to collect jejunum to investigate morphological changes and cecal contents to determine microbial population. The data were analyzed using ANOVA procedures of SAS 9.4. The means were compared by Duncan’s multi-range test at P < 0.05 level of significance. Rats in SPL were heavier than those in CON by 15.1 % at day 16. Jejunal villus height in SPL was highest among treatments at day 8, and it was also significantly lengthened in SPL than other treatments at day 16 (P < 0.05). Cecal populations of E. coli, Streptococcus spp. and Salmonella spp. were significantly reduced by 64.7 %, 67.2 % and 40.0 % in SPL, respectively, relative to CON at day 16 (P < 0.05). Collectively, SPL accelerated growth by improving intestinal restoration and modulating microbiota population. SPL might have a potential as a feed additive to reduce early-weaning stress of livestock animals.

Production and characterization of sophorolipids by Candida bombicola using catfish fat

Catfish fat is a high nutrition by-product of seafood processing industry. In this work, catfish fat has been used as a sustainable and economical raw material for sophorolipids production by Candida bombicola. Sophorolipids yield was maximum as 21.8g/L after 7 days of fermentation at 25oC, pH 6, 180 rpm. The obtained sophorolipids was to contained the main component as lactonic sophorolipids, which has been confirmed by Thin layer chromatography (TLC). Sophorolipids also exhibited the ability to resistant Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and showed the ability of free radical scavenging the dose dependent manner with IC50 was 4.45 mg/ml. These results suggested that sophorolipids could be used in health care products and cosmetic. Catfish fat could be used as the low cost hydrophobic carbon source to replace fatty acid for sophorolipids production.

Rational high-throughput screening system for high sophorolipids production in Candida bombicola by co-utilizing glycerol and glucose capacity

BackgroundA rational and high-performance high-throughput screening system effectively improves the efficiency of strain screening.ResultsA rapid and efficient method was developed to detect carbon sources. And then by using glucose analog as screening pressure and two indexes for sophorolipids (SLs) production and glycerol consumption, a mutant of Candida bombicola L1.3 was successfully selected among 3000 mutants. It produces high SLs titer and co-utilizes glucose and glycerol simultaneously.ConclusionsCompared to the wild-type strain, L1.3 exhibited 15.06% higher SLs titer and 35.69% higher glycerol consumption capacity in a 5-L bioreactor. We believe that L1.3 can potentially be used for the efficient industrial production of SLs with simple downstream processing for separating SLs and glycerol.

Application of biosurfactants and chitosan in toothpaste formulation

The aim of the present study was to formulate toothpastes containing biosurfactants and either fungal chitosan or sodium fluoride and evaluate the cytotoxicity, antimicrobial action and inhibition potential against biofilm formed by Streptococcus mutans. Chitosan was extracted from the biomass of the fungus Mucorales. We tested biosurfactants produced by Pseudomonas aeruginosa UCP 0992 (PB), Bacillus metylotrophicus UCP 1616 (BB) and Candida bombicola URM 3718 (CB). Fractional inhibitory concentration analysis was performed to determine the type of interaction between the compounds. Six toothpaste were prepared, the active ingredients of which were the biosurfactants, chitosan or sodium fluoride. The cytotoxicity tests were performed using the 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay for the L929 (mouse fibroblast) and RAW 264.7 (mouse macrophage) cell lines. The toothpastes were tested with regard to pH, consistency and foaming capacity. The inhibition of biofilm was investigated by applying the toothpaste to biofilm formed in modified artificial saliva for 24 h at 37 °C in anaerobiosis. All substances had a minimum inhibitory concentration (MIC) for S. mutans. The combinations of CB and PB with chitosan had an additive effect against S. mutans, whereas BB combined with chitosan had an indifferent effect. The toothpastes were non-toxic. The formulations had pH around 9, spreading capacity between 8 and 17 mm and foaming capacity between 63 and 95%. All formulations inhibited the cellular viability of S. mutans in the biofilm, with similar results compared to the commercial toothpaste tested. The present results show that the formulations suggested are promising when compared to a commercial tooth paste.

Formulation and evaluation of wound healing activity of sophorolipid-sericin gel in wistar rats

Sericin is a useful by-product of silk processing and is resistant to oxidation and ultraviolet and can absorb and release moisture easily. Sericin has biological activities such as antibacterial, antioxidant, and tyrosinase inhibition. Sophorolipids are microbial extracellular glycolipids produced by resting cells of Candida bombicola. Sophorolipids show excellent skin compatibility and also have antibacterial property. In this study, we have developed a novel formulation consisting of sericin and sophorolipid with calcium alginate as a binding agent. Since both the ingredients are biocompatible and biodegradable, the formulation was tested for wound healing in Wistar rats. A commercial ointment povidone was used as control. The animal group, treated with sericin and sophorolipid cream, showed fast contraction, rapid closure, and healing when compared with control and commercial ointment. These observations were validated with histopathological studies where more fibroblast proliferation, angiogenesis, and keratinization were observed. This is a green, cost-effective formulation for fast wound healing.

Sophorolipid Biosurfactants Activate Taste Receptor Type 1 Member 3‐Mediated Taste Responses and Block Responses to Bitter Taste In Vitro and In Vivo

AbstractSophorolipids (SL) are typically produced and secreted by select nonpathogenic yeast species (i.e., Candida) from renewable substrates. They are currently being used by industry on a limited basis in formulations for cleaning solutions as well as laundry and dishwashing detergents. Due to the nature of their chemical structure, it was hypothesized that SL would demonstrate taste‐sensory properties. In this study, SL were produced via fermentation on a mixed substrate platform with glucose and either palmitic acid, stearic acid, or oleic acid using Candida (currently reclassified as Starmerella) bombicola ATCC 22214. The taste properties of SL were determined using a single‐cell manual calcium imaging technique on cultured human fungiform taste papillae (HBO) cells. The results of those studies demonstrated that sweetener‐responsive HBO cells also respond to SL, and these responses are mediated by the type 1 taste receptors 3 (T1R3), because they were blocked by lactisole (a T1R3 receptor‐specific blocker). The involvement of the T1R3 receptor in SL recognition was confirmed via the chorda tympani nerve recording (CTNR) study in a (−/−) T1R3 knockout (KO) mouse model. We further demonstrated that SL are capable of blocking the bitter stimuli‐elicited responses both in HBO cells and in the CTNR study. This is the first report demonstrating that SL have taste‐sensory properties, which opens up numerous possibilities for practical applications of SL to ameliorate bitter tastes in foods and drugs and understand the potential source of dysgeusia in some patients.

Mouthwash containing a biosurfactant and chitosan: An eco-sustainable option for the control of cariogenic microorganisms.

The aim of the present study was to determine the antimicrobial action and toxicity of mouthwashes formulated with a biosurfactant, chitosan of a microbial origin and peppermint (Mentha piperita) essential oil (POE). Chitosan was extracted from the biomass of a fungus from the order Mucorales grown in yam bean broth. Three biosurfactants produced by Pseudomonas aeruginosa UCP 0992 (PB), Bacillus cereus UCP 1615 (BB) and Candida bombicola URM 3718 (CB) were tested. Six mouthwashes were prepared, the active ingredients of which were the biosurfactant, chitosan and POE. The minimum inhibitory concentration (MIC) was determined for the test substances separately, in combinations and in the mouthwash formulas. The toxicity of the mouthwashes was tested using MTT (3-(4,5-dimethylthiazole-2-il)-2,5-diphenyltetrazolium bromide) for the L929 (mouse fibroblast) and RAW 264.7 (mouse macrophage) cell lines. All substances tested had a MIC for cariogenic microorganisms. The combinations of the CB and PB biosurfactants with chitosan demonstrated an additive effect on the majority of microorganisms tested. The toxicity of the mouthwashes was significantly lower than that of the commercial mouthwash. The present findings demonstrate that mouthwashes containing natural products constitute a safe, effective, natural alternative to commercially available mouthwashes for the control of oral microorganisms.

Metabolic profiling and flux distributions reveal a key role of acetyl-CoA in sophorolipid synthesis by Candida bombicola

Sophorolipids are important biosurfactants produced by the wild-type strain of Candida bombicola ATCC 22214, with antimicrobial, antiviral, and anticancer activities. Our previous study showed that a mutant strain lacking multifunctional enzyme type 2 displayed extremely low sophorolipid production capacity compared to the wild-type strain. In the present study, we aimed to reveal the possible reasons for this deficiency through a series of analytical methods. First, comparative metabolic profiling analysis between the wild-type and mutant strains was performed. Multivariate statistical analysis, including principal component analysis and orthogonal partial least squares discriminant analysis, showed that nine potential metabolites were closely associated with the metabolic differences between strains. Second, metabolic flux analysis revealed that an inadequate supply of intracellular acetyl-CoA in the mutant strain could significantly affect the biosynthesis of sophorolipids. In addition, supplementation of citric acid recovered the sophorolipid production in the mutant strain by approximately 83%, suggesting that citrate metabolism plays an important role in sophorolipid biosynthesis. These findings provide novel theoretical insights for further improving the production of sophorolipids in C. bombicola.

Rational high-throughput system for screening of high sophorolipids-producing strains of Candida bombicola

A rational high-throughput screening can significantly improve the efficiency of strain screening with high performance. In this study, based on the addition reaction of unsaturated fatty acids in the sophorolipids (SLs) and I2 molecules, a simple and rapid high-throughput detection method for SLs was established which demonstrated a correlation coefficient (R2) of 0.9106 with high-performance liquid chromatography (HPLC) method. Moreover, chlorpromazine, as a rational selecting pressure for enrichment of mutants with high cytochrome P450 enzyme activity, which was a key enzyme in the synthesis of SLs, was introduced into the high-throughput screening model. Consequently, with the aid of this effective screening system, a high-yielding mutant designated as Candida bombicola F6.5 was successfully screened out from 1500 single colonies, which presented improvements of 40.3% and 11.4% on SLs titer and yield, respectively, compared to the parent strain in a 1L bioreactor.

Bioleaching of trace elements and rare earth elements from coal fly ash

Coal fly ash originated from coal combustion has high concentrations of metals. If suitable leaching techniques are identified, then coal fly ash could serve as a useful source of valuable minerals including rare earth elements (REEs). In this study, three microbial strains, Candida bombicola, Phanerochaete chrysosporium and Cryptococcus curvatus were tested on their performance of leaching trace elements and REEs from fly ash. Through comparing mineral loss and leaching efficiencies resulting from indirect leaching or use of the culture supernatant, C. bombicola was identified to be the best leading to the highest mineral loss and extracting efficiencies of trace elements and REEs among the three strains. The highest mineral loss observed from using the supernatant of this yeast strain was 59.7%. Among all trace elements, As and Mo had the highest leaching efficiency of 80.9% and 79.5%, respectively. The same leaching test led to 67.7% of Yb and 64.6% of Er dissolved from the ash. This study, thus, demonstrated that bioleaching is feasible for leaching metals out of fly ash. The C. bombicola strain deserves further investigation due to its robust actions on metal leaching.

Antiangiogenic activity of sophorolipids extracted from refined bleached deodorized palm olein.

OBJECTIVES:Sophorolipids (SLs) are a group of surface-active glycolipids produced by a type of nonpathogenic yeast Candida bombicola in the presence of vegetable oil through fermentation technology. SLs have shown antitumor activity; however, the mechanism of action underlying the anticancer activity of SLs is poorly understood. This work evaluated the anticancer activity of SLs fermented from palm oil by exploring its antiangiogenic activity.MATERIALS AND METHODS:The SLs that were fermented and further characterized for their biochemical activities. Cytotoxicity study was performed to assess cytostatic properties. A series of in vitro and ex vivo angiogenesis assay was also carried out. The relative fold change in the expression of p53 mRNA by SLs was also studied.RESULTS:Altogether, the data show that SLs derived from palm oil fermentation process inhibited neovascularization in the ex vivo tissue segments and also the endothelial cell proliferation between 50% and 65% inhibition as a whole. The palm oil derived SLs also caused downregulation of the suppression level of vascular endothelial growth factor and also upregulate the p53 mRNA level. The analytical studies revealed the presence of high amount of phenolic compounds but with relatively weak antioxidant activity. The gas chromatography-mass spectrometry studies revealed abundant amount of palmitic and oleic acid, the latter an established antiangiogenic agent, and the former being proangiogenic.CONCLUSION:Therefore, it can be concluded from this study that SLs derived from fermented palm oil have potent antiangiogenic activity which may be attributed by its oleic acid component.

Investigation of fermentation conditions for Candida bombicola ACTT22214 from molasses and soybean oil for sophorolipid production

Sophorolipid (SL) is a biosurfactant belonging to the glycolipids group and was produced by harmless Candida bombicola ACTT22214 and has been widely used in many fields in our life. In order to search for appropriate condition for C. bombicola fermentation producing SL with high efficiency, this study focused on the investigation of the soy oil concentration, molasses concentration, fermentation time, pH and temperature. SL products were tested for antimicrobial activity, antioxidant, emulsifier, foaming ability. The highest content of SL was 43.27 ± 0.30 g/L under conditions of: soybean oil content 5%, molasses content 150 g/L, fermentation time 7 days, pH = 5, 28oC fermentation temperature. The antibacterial activity of SL was good: the highest resistance to Candida albicans (16.33 ± 1.15 mm), good resistance to Bacillus spizizenii (13.67 ± 0.58 mm), resistance to Staphylococcus aureus (12.67 ± 1.15 mm), relatively weak resistance to Pseudomonas aeruginosa (11.33 ± 0.58 mm) and Escherichia coli (9.67 ± 0.58 mm). The antioxidant capacity of SL was quite high with an IC50 value of 6.024 mg/mL. The emulsifying capacity of SL was equivalent to the emulsification of the tween 20 at a concentration of 5 - 10 mg/mL. SL had the ability to foam evenly from concentrations of 5 to 20 mg/mL but not higher than the corresponding concentrations of tween 20, SL was smooth, even, stable longer than tween 20

Antimicrobial applications of sophorolipid from Candida bombicola: A promising alternative to conventional drugs

Antimicrobial applications of sophorolipid from Candida bombicola: A promising alternative to conventional drugs

A novel microbial technique for producing high-quality sophorolipids from horse oil suitable for cosmetic applications.

SummaryHorse oil contains linoleic, palmitoleic and unsaturated fatty acids that are similar to those in human skin, and may therefore be an ideal substance from which to isolate biosurfactants for cosmetic products to improve human skin quality. Herein, an innovative approach was developed to synthesise sophorolipids from horse oil by hydrolysis, followed by fermentation using the yeast Candida bombicola. The yield of sophorolipids from direct fermentation of horse oil and hydrolysed horse oil was 40.6 ± 1.3 g l−1 and 58.4 ± 1.8 g l−1 respectively. To further increase the yield, 30–40 g l−1 glucose was added in a fed‐batch fermentation process to maintain the pH between 4.0 and 4.5, resulting in a conversion yield of 71.7 ± 0.8 g l−1. The purity and structure of the synthesised sophorolipids were analysed by ultra‐performance liquid chromatography‐mass spectrometry and nuclear magnetic resonance. An in vitro human dermal fibroblast model was used as a surrogate for human skin to measure elastase inhibition activity. Antiwrinkle properties of isolated sophorolipids were better than those of horse oil or hydrolysed horse oil in several in vitro assays. Furthermore, no cytotoxicity was observed at a concentration of 50 μg ml−1, and wound‐healing capacity was evident in a cell culture model. Additionally, the synthesised sophorolipids attenuated lipopolysaccharide‐induced expression of inflammatory cytokines in macrophages, and efficiently inhibited several strains of bacteria and yeast. In conclusion, fed‐batch fermentation of hydrolysed horse oil is a novel and efficient approach for producing high‐quality and high‐yield sophorolipids that exhibit great potential as cosmetic ingredients.

Antimicrobial Applications of Sophorolipid from Candida bombicola: a Promising Alternative to Conventional Drugs

Antimicrobial Applications of Sophorolipid from Candida bombicola: a Promising Alternative to Conventional Drugs

Hypocholesterolemic impact of newly isolated sophorolipids produced by microbial conversion of safflower oil cake in rats fed high-fat and cholesterol diet

The present study aims to produce low cost sophorolipids, and to evaluate their potential hypocholesterolemic impact. Sophorolipids were produced by Candida bombicola grown on safflower oil cake, extracted by methanol followed by ethyl acetate with a yield of 24.4 and 48.3 g·100 g-1 mixed substrate, respectively. Their structure was confirmed by FTIR and 1H NMR and proven to be safe when subjected to an acute toxicity test. A biological experiment was done on 42 male albino rats classified into six groups for 4 weeks following an induction period for hypercholesterolemia of 8 weeks. The two extracts and their mixture were examined for their hypocholesterolemic effect compared to rosuvastatin. The results revealed a reduction in total cholesterol, low density lipoprotein cholesterol, atherogenic index, liver transaminases’ activity and malondialdehyde. They also revealed an elevation in high density lipoprotein cholesterol and antioxidant enzymes which was more efficient than rosuvastatin. Histopathological examination confirmed these results. In conclusion, the newly isolated sophorolipids are powerful hypocholesterolemic compounds which are even more efficient and safer than rosuvastatin.

Lubricant Properties of ω − 1 Hydroxy Branched Fatty Acid-Containing Natural and Synthetic Lipids

Hydroxy fatty acids, such as those derived from castor and lesquerella seed oils, make ideal substrates for the synthesis of bio-lubricants, cosmetics, coatings, plastics, and lubricant additives. However, feedstocks of such fatty acids suffer from major drawbacks, such as a lack of a cropping system to produce those seeds or toxic by-products in generating the seed oil, all of which limit availability and thus add to costs. In this study, we explore lubrication properties of microbially derived hydroxy fatty acids and demonstrate that such microbial ω − 1 hydroxy fatty acids, and their derivatives, exhibit lubrication traits (e.g., anti-friction and anti-wear properties) comparable to those of seed-derived hydroxy fatty acids. These ω − 1 hydroxy fatty acids can be recovered from sophorolipids produced by the yeast Candida bombicola ATCC 22214, or by bioengineering bacterial systems to produce them from sugar (Garg et al. in Metab Eng 35:9–20, 2016). Optimization of this latter system can pave the way for a less costly and sustainable alternative to plant-derived bio-lubricants.

Fundamental Characterization of the Micellar Self-Assembly of Sophorolipid Esters.

Surfactants are ubiquitous constituents of commercial and biological systems that function based on complex structure-dependent interactions. Sophorolipid (SL) n-alkyl esters (SL-esters) comprise a group of modified naturally derived glycolipids from Candida bombicola. Herein, micellar self-assembly behavior as a function of SL-ester chain length was studied. Surface tensions as low as 31.2 mN/m and critical micelle concentrations (CMCs) as low as 1.1 μM were attained for diacetylated SL-decyl ester (dASL-DE) and SL-octyl ester, respectively. For deacetylated SL-esters, CMC values reach a lower limit at SL-ester chains above n-butyl (SL-BE, 1-3 μM). This behavior of SL-esters with increasing hydrophobic tail length is unlike other known surfactants. Diffusion-ordered spectroscopy (DOSY) and T1 relaxation NMR experiments indicate this behavior is due to a change in intramolecular interactions, which impedes the self-assembly of SL-esters with chain lengths above SL-BE. This hypothesis is supported by micellar thermodynamics where a disruption in trends occurs at n-alkyl ester chain lengths above those of SL-BE and SL-hexyl ester (SL-HE). Diacetylated (dA) SL-esters exhibit an even more unusual trend in that CMC increases from 1.75 to 815 μM for SL-ester chain lengths of dASL-BE and dASL-DE, respectively. Foaming studies, performed to reveal the macroscopic implications of SL-ester micellar behavior, show that the observed instability in foams formed using SL-esters are due to coalescence, which highlights the importance of understanding intermicellar interactions. This work reveals that SL-esters are an important new family of green high-performing surfactants with unique structure-property relationships that can be tuned to optimize micellar characteristics.

Sweet sorghum bagasse and corn stover serving as substrates for producing sophorolipids.

To make the process of producing sophorolipids by Candida bombicola truly sustainable, we investigated production of these biosurfactants on biomass hydrolysates. This study revealed: (1) yield of sophorolipds on bagasse hydrolysate decreased from 0.56 to 0.54 and to 0.37g/g carbon source when yellow grease was dosed at 10, 40 and 60g/L, respectively. In the same order, concentration of sophorolipids was 35.9, 41.9, and 39.3g/L; (2) under similar conditions, sophorolipid yield was 0.12, 0.05 and 0.04g/g carbon source when corn stover hydrolysate was mixed with soybean oil at 10, 20 and 40g/L. Sophorolipid concentration was 11.6, 4.9, and 3.9g/L for the three oil doses from low to high; and (3) when corn stover hydrolysate and yellow grease served as the substrates for cultivating the yeast in a fermentor, sophorolipid concentration reached 52.1g/L. Upon further optimization, sophorolipids production from ligocellulose will be indeed sustainable.

Production and characterization of sophorolipids produced by Candida bombicola from coconut oil

The biosurfactants from microbial origin increasingly gained interests because of their application in many field and excellent properties compared to surfactants from chemical origin, such as the higher biodegradability, lower toxicity and environmentally friendly. Sophorolipids, biosurfactants of glycolipid groups are produced through the fermentation by nonpathogenic yeasts such as Candida bombicola. In this study, we investigated the production, surveyed properties of sophorolipids through fermentation by C. bombicola from coconut oil. The results showed that the yield of sophorolipid obtained after 7 days of culture was 14.6 g/L, the surface tension was 40 mN/m. The obtained sophorolipid showed ability to be resistant to some bacteria such as E. coli, B. subtilis, P. aeruginosa, and S. aureus. Through DPPH experiment, sophorolipids showed the scavenging acitivity with IC50 = 1.4063 mg/mL. These results showed that sophorolipids could be applied in cosmetics.