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To develop a structural homolog of mannosylerythritol lipids (MELs), Pseudozyma tsukubaensis JCM16987 (known to be a specific producer of the diastereomer type of mono-acetylated MEL (MEL-B)) was cultivated in medium containing 4 % (w/v) olive oil as the primary carbon source and 4 % L-arabitol as the supplemental sugar alcohol. Based on thin-layer chromatography (TLC), the glycolipid extract showed two major spots corresponding to MEL-B and an unknown glycolipid (GL1). Based on high-performance liquid chromatography after acid hydrolysis, GL1 from the L-arabitol culture showed two primary peaks identical to mannose and arabitol using the sugar analysis column, and one peak identical to L-arabitol was detected using the chiral resolution column. Based on NMR analysis, GL1 was identified as mono-acetylated mannosyl-L-arabitol lipid (MLAL-B) consisting of mannose, with L-arabitol as the sugar moiety. The observed critical micelle concentration (CMC) and surface tension at the CMC (γCMC) of MLAL-B were 1.2 × 10(-5) M and 32.8 mN/m, which were significantly higher than MEL-B (CMC = 3.1 × 10(-6) M and γcmc = 26.1 mN/m). Furthermore, based on a water-penetration scan, MLAL-B efficiently formed lamellar phase (Lα) and myelins at a broad concentration range. Thus, the present glycolipid showed higher hydrophilicity and/or water solubility and increased our understanding of environmentally advanced biosurfactants.

The spreading property of mannosylerythritol lipids (MELs) was investigated in connection with our search for new application in agriculture. The wetting ability of MEL solutions for hydrophobic surfaces was evaluated based on contact angle measurements for several surfactant solutions on abiotic and biotic surfaces. The contact angle of MEL-A solution on a hydrophobic plastic surface at 100 s after placement decreased to 8.4°, and those of other MEL solutions decreased more significantly compared to those of commonly-used nonionic surfactants. In addition, the contact angle of MEL solutions also dropped down to around 10° on various plant leaf surfaces. MEL solutions, in particular, efficiently spread even on poorly wettable Gramineae plant surfaces on which general nonionic surfactant solutions could not. Moreover, the wetting ability of MEL solutions was found to be greatly affected by the structural difference in their carbohydrate configuration. Furthermore, surface pretreatment with MEL solution led to more efficient spreading and fixing of microbial cells onto plant leaf surface compared to several conventional surfactants used in this study. These results suggested that MELs have a potential to use as a natural bio-based spreading agent, particularly as agrochemical spreader for biopesticides.

The synergic effect and miscibility of the lactonic and acidic forms of sophorolipids (SLs) produced by Starmerella bombicola NBRC 10243 were first evaluated through atomic force microscopy (AFM), together with the Langmuir monolayer technique. The π-A isotherm of a pure lactonic sophorolipid (LS) monolayer mostly exhibited a liquid expanded monolayer, while that of an acidic sophorolipid (AS) monolayer showed a liquid condensed monolayer, suggesting that the lactonization of SLs makes the molecules more bulky and prevents them from adopting a close-packed arrangement. Plots of the mean area per molecule of mixed LS/AS monolayers gave positive deviations from the ideal curves, implying that the LS and AS molecules are miscible. Interestingly, the positive deviation of excess area (Aex) from ideality was most significant at a mole fraction (XAS) of 0.3, which closely resembles the composition of the LS/AS mixture secreted by S. bombicola in culture. The AFM images of mixed LS/AS monolayers transferred at 20 mN/m revealed no phase-separated microdomain structures, but rather showed small protruding objects for all compositions, indicating that LS and AS are partially miscible, as predicted by the positive deviations from the ideal curves. Cross-section analysis of the AFM images indicated that the observed protruding objects are AS-rich monolayers formed on the LS/AS monolayer. Our results clearly demonstrate that AFM combined with the Langmuir technique is useful for the exploration of the miscibility and synergic effects of microbial products.

Biosurfactants are surface-active compounds produced by microorganisms. Mannosylerythritol lipids (MEL) are promising biosurfactants produced by Ustilaginomycetes, and their physicochemical and biochemical properties differ depending on the chemical structure of their hydrophilic and/or hydrophobic moieties. To further develop MEL derivatives and expand their potential applications, we focused our attention on the use of cuttlefish oil, which contains polyunsaturated fatty acids (e.g., docosahexaenoic acid, C₂₂:₆, and eicosapentaenoic acid, C₂₀:₅, as the sole carbon source. Among the microorganisms capable of producing MEL, only nine strains were able to produce them from cuttlefish oil. On gas chromatography-mass spectrometry (GC/MS) analysis, we observed that Pseudozyma churashimaensis OK96 was particularly suitable for the production of MEL-A, a MEL containing hexadecatetraenoic acid (C₁₆:₄) (23.6% of the total unsaturated fatty acids and 7.7% of the total fatty acids). The observed critical micelle concentration (CMC) and surface tension at CMC of the new MEL-A were 5.7×10⁻⁶ M and 29.5 mN/m, respectively, while those of MEL-A produced from soybean oil were 2.7×10⁻⁶ M and 27.7 mN/m, respectively. With polarized optical and confocal laser scanning microscopies, the self-assembling properties of MEL-A were found to be different from those of conventional MEL. Furthermore, based on the DPPH radical-scavenging assay, the anti-oxidative activity of MEL-A was found to be 2.1-fold higher than that of MEL-A produced from soybean oil. Thus, the newly identified MEL-A is attractive as a new functional material with excellent surface-active and antioxidative properties.

Mannosylerythritol lipids (MELs) produced by yeasts are one of the most promising glycolipid biosurfactants. There are two MEL diastereomers, in which the configurations of the erythritol moieties are opposite. The 4-O-β-D-mannopyranosyl-(2S,3R)-erythritol (S-form) or 4-O-β-D-mannopyranosyl-(2R,3S)-erythritol (R-form) is the hydrophilic domain. In this study, we prepared S- and R-form MEL homologs with similar fatty acyl groups, and compared their interfacial properties. Among the four diastereomers (S-MEL-B and -D/R-MEL-B and -D), R-form MELs showed a higher critical aggregation concentration and hydrophilicity compared to the corresponding S-form. R-form MELs also efficiently formed relatively large vesicles compared to S-form. Moreover, we estimated the binary phase diagram of the MEL-water system and compared the aqueous phase behavior among the four diastereomers. The present MELs self-assembled into a lamellar (L(α)) structure at all concentration ranges. Meanwhile, the one-phase L(α) region of R-form MELs was wider than those of S-form MELs. R-form MELs may maintain more water between the polar layers in accordance with an extension of the interlayer spacing. These results suggest that the differences in MEL carbohydrate configurations significantly affect interfacial properties, self-assembly, and hydrate ability., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Mannosylerythritol lipids (MELs) are secreted by yeasts and are promising glycolipid biosurfactants. In our study on the non-aqueous phase behaviors of MEL homologues, we found that MEL-D (4-O-[2',3'-di-O-alka(e)noyl-β-D-mannopyranosyl]-(2R,3S)-erythritol) forms aggregates in decane. The microscopic observation and the X-ray scattering measurement of these aggregates revealed that they are reverse vesicles that consist of bilayers whose hydrophilic domains are located in the interior of the bilayers. In addition, MEL-D formed reverse vesicles without co-surfactants and co-solvents in various oily solutions, such as n-alkanes, cyclohexane, squalane, squalene, and silicone oils at a concentration below 10 mM. This is the first report on the reverse vesicle formation from biosurfactants.

Mannosylerythritol lipids (MELs) produced by yeasts are one of the most promising glycolipid biosurfactants. In this study, we succeeded in the preparation of a novel MEL homolog having no acetyl groups, namely MEL-D. MEL-D was synthesized by lipase-catalyzed hydrolysis of acetyl groups from a known MEL, and identified as 4-O-[2',3'-di-O-alka(e)noyl-β-d-mannopyranosyl]-(2R,3S)-erythritol. The obtained MEL-D showed a higher critical aggregation concentration (CAC=1.2 × 10(-5)M) and hydrophilicity compared to known MELs, retaining an excellent surface tension lowering activity (the surface tension at the CAC was 24.5mN/m). In addition, we estimated the binary phase diagram of the MEL-D-water system based on a combination of visual inspection, polarized optical microscopy, and SAXS measurement. From these results, MEL-D was found to self-assemble into a lamellar (L(α)) structure over all ranges of concentration. Meanwhile, the one-phase L(α) region of MEL-D was extended wider than those of known MELs. MEL-D might keep more water between the polar layers in accordance with the extension of the interlayer spacing (d). These results suggest that the newly obtained MEL-D would facilitate the application of MELs in various fields as a lamellar-forming glycolipid with higher hydrate ability., (2010 Elsevier Ltd. All rights reserved.)

Cryptococcus humicola JCM 1461 efficiently produced cellobiose lipids (CLs), bolaform biosurfactants. The main product was identified as 16-O-(2″,3″,4″,6'-tetra-O-acetyl-β-cellobiosyl)-2-hydroxyhexadecanoic acid. The production yield of CLs reached 13.1 g/L under the intermittent feeding of glucose. The critical micelle concentrations (CMC) of the main product at pH 4.0 and 7.0 were 3.3×10(-5) M and 4.1×10(-4) M respectively.

Mannosylerythritol lipids (MELs) are glycolipid biosurfactants excreted by fungal strains. They show not only excellent surface-active properties but also versatile biochemical actions. Ustilago scitaminea NBRC 32730 has been reported mainly to produce a mono-acetylated and di-acylated MEL, MEL-B, from sucrose as sole carbon source. In order to make biosurfactant production more efficient, we focused our attention on the use of sugarcane juice, one of the most economical resources. The fungal strain produced MEL-B at the yield of 12.7 g/L from only sugarcane juice containing 22.4% w/w sugars. Supplementation with organic (yeast extract, peptone, and urea) and inorganic (sodium nitrate and ammonium nitrate) nitrogen sources markedly enhanced the production yield. Of the nitrogen sources, urea gave the best yield. Under optimum conditions, the strain produced 25.1 g/L of MEL-B from the juice (19.3% sugars) supplemented with 1 g/L of urea in a jar fermenter at 25 °C over 7 d. The critical micelle concentration (CMC) and the surface-tension at the CMC for the present MEL-B were 3.7×10(-6) M and 25.2 mN/m respectively. On water-penetration scan, the biosurfactant efficiently formed the lamella phase (L(α)) and myelins over a wide range of concentrations, indicating excellent surface-active and self-assembling properties. More significantly, the biosurfactant showed a ceramide-like skin-care property in a three-dimensional cultured human skin model. Thus, sugarcane juice is likely to be effective in glycolipid production by U. scitaminea NBRC 32730, and should facilitate the application of MELs.

Sophoroselipids (SL) are bolaform biosurafactants which are abundantly produced by microorganisms from renewable resources. In this study, four kinds of bolaform biosurfactants were produced, and these derivatives were chemoenzymatically synthesized from "acid form" diacetylated SL (SLdiAc), which are preferentially produced by Candida floricola TM 1502 which was newly found in our group. The effects of the structure of sugar moiety on their surface-active properties were investigated by surface tension measurement. After microbial production of SLdiAc from oleic acid/glucose, SLdiAc was converted into acetylated glucoselipid (GLAc). Among twelve species of glucosidases, pectinase and pectolyase including polygalacturonase were found to cleave the beta-1,2-glycosidic linked disaccharide, especially pectolyase produced GLAc effectively at 40 degrees C and pH 4.0. The structure of the major component of purified GLAc was assigned as 17- [(beta-D-glucopyranosyl)oxy]-cis-9-octadecenoate 6'-acetate by using NMR analyses, MALDI-TOF/MS and GC-MS. Glucoselipid (GL) without acetyl group was also enzymatically converted from SL obtained from alkaline hydrolysis of SLdiAc. Interestingly, the estimated CMC values of SLdiAc, SL, GLAc, GL indicated almost the same values despite their difference in hydrophilic structure. Although the difference in monosaccaride and disaccharide also did not affect gammaCMC, the presence of acetyl group on sugar moiety was found to lower the gammaCMC value slightly, suggesting that the acetyl group on produced bolaform biosurfactant is likely to play more important role to reduce the free energy of air/water interface.

Glycolipid biosurfactants, mannosylerythritol lipids (MELs), were produced from glucose and sucrose without vegetable oils. Pseudozyma antarctica JCM 10317, Ustilago maydis NBRC 5346, U. scitaminea NBRC 32730, and P. siamensis CBS 9960 produced mainly MEL-A, MEL-A, MEL-B, and MEL-C respectively. The sucrose-derived MELs showed excellent interfacial properties: low critical micelle concentration as well as that of oil-derived MELs.

A smut fungus Ustilago scitaminea NBRC 32730 on sugar cane (Saccharum) was found to accumulate a large amount of glycolipids in the culture medium. As a result of structural characterization, the main glycolipid was identified as MEL-B, 4-O-beta-(2',3'-di-O-alka(e)noyl-6'-O-acetyl-D-mannopyranosyl)-erythritol. The MEL-B was sufficiently produced from a variety of sugars such as sucrose, glucose, fructose, and mannose. Olive oil and methyl oleate were also available as carbon sources to produce MEL-B. However, these residual oils made product recovery very complicated. Under optimal conditions, a maximum MEL yield of 12.8 g/l was achieved by feeding of sucrose.

Mannosylerythritol lipids (MELs) are glycolipid biosurfactants (BS) abundantly produced from renewable resources by yeast strains of the genus Pseudozyma. In this study, the ternary phase behaviors of two types of MELs, i.e. MEL-A and MEL-B, mixed with water and oil were investigated at 25 degrees C based on polarized optical microscopy and small-angle X-ray scattering (SAXS). When n-decane was used as an oil phase, diacetylated MEL-A formed single-phase water-in-oil (W/O) microemulsion in a remarkably large region. MEL-A, with a negative spontaneous curvature, also formed sponge (L(3)), reverse bicontinuous cubic (V(2)), and lamellar (L(alpha)) phases. Meanwhile, monoacetylated MEL-B, with the opposite configuration of the erythritol moiety, gave single-phase bicontinuous microemulsion and showed a triangular phase diagram dominated by the L(alpha) phase, suggesting that MEL-B has an almost zero spontaneous curvature. Moreover, we succeeded in preparation of oil-in-liquid crystal (O/LC) emulsion in the biphasic L(alpha)+O region of the MEL-B/water/n-decane system. The obtained gel-like emulsion was stable for at least 1 month. These results clearly demonstrated that the difference in the number of acetyl group on the headgroup and/or the chirality of the erythritol moiety drastically changed the phase behavior of MELs. Accordingly, these MELs would be quite distinctive from conventional BS hitherto reported, and would have great potential for the preparation of microemulsion and LC-based emulsion.

Mannosylerythritol lipids (MELs) are glycolipid biosurfactants produced by the yeast strains of the genus Pseudozyma. These compounds show not only excellent surface-active properties, but also versatile biochemical actions. During a survey of new MEL producers, we found that a basidiomycetous yeast, Pseudozyma crassa, extracellularly produces three glycolipids. When glucose and oleic acid were used as the carbon source, the total amount of glycolipids reached approximately 4.6g/L in the culture medium. The structures of these glycolipids were similar to those of well-known MEL-A, -B, and -C, respectively. Very interestingly, in all the present glycolipids, the configuration of the erythritol moiety was entirely opposite to that of conventional MELs. The present glycolipids were identified to have the carbohydrate structure of 4-O-beta-D-mannopyranosyl-(2R,3S)-erythritol, stereochemically different from 4-O-beta-D-mannopyranosyl-(2S,3R)-erythritol of conventional MELs. Furthermore, these new glycolipids possessed both short-chain acids (C(2) or C(4)) and long-chain acids (C(14), C(16), or C(18)) on the mannose moiety. The major component of the present glycolipids clearly showed different interfacial and biological properties, compared to conventional MELs comprising two medium-chain acids on the mannose moiety. Accordingly, the novel MEL diastereomers produced by P. crassa should provide us with different glycolipid functions, and facilitate a broad range of applications of MELs.

Mannosylerythritol lipids (MELs) are one of the most promising glycolipid biosurfactants produced by yeast strains of the genus Pseudozyma. In this study, the aqueous-phase behavior of a new monoacetyl MEL derivative, 1-O-beta-(2',3'-di-O-alka(e)noyl-6'-O-acetyl-d-mannopyranosyl)-d-erythritol (MEL-B), was investigated using polarized optical microscopy, small-angle X-ray scattering (SAXS), confocal laser scanning microscopy (CLSM), and differential scanning calorimetry (DSC). The present MEL-B was found to self-assemble into a lamellar (L(alpha)) phase over remarkably wide concentration and temperature ranges. According to SAXS measurement, the interlayer spacing (d) was estimated to be almost constant (about 4.7 nm) at the low MEL-B concentration (60 wt.%) region, the d-spacing gradually decreased to 3.1 nm with an increase in the MEL-B concentration. The thermal stability of the liquid crystalline phase was investigated by DSC measurement. The obtained L(alpha) phase was found to be stable up to 95 degrees C below a MEL-B concentration of 85 wt.%; then, the melting temperature of the liquid crystalline phase dramatically decreased with an increase in MEL-B concentration (above 85 wt.%). Furthermore, we found relatively large vesicles (1-5 microm) at the low MEL-B concentration using CLSM observation. The trapped volume of the obtained MEL-B vesicle was estimated to be about 0.42 microL/mumol by glucose dialysis method. These results suggest that the natural glycolipid biosurfactant, the newly found MEL-B, would be useful in various fields of applications as an L(alpha) phase- and/or vesicle-forming lipid.

Mannosylerythritol lipid-A (MEL-A) is a glycolipid biosurfactant abundantly produced from soybean oil by microorganisms at a yield of up to 100 g/L. In this study, the formation of water-in-oil (W/O) microemulsion based on the single component of MEL-A was confirmed using dynamic light scattering (DLS) and freeze fracture electron microscopy (FF-EM). DLS and FF-EM measurements revealed that the diameter of the microemulsion increases with an increase in water-to-surfactant mole ratio (W(0)) ranging from 20 to 60 nm, and the maximum W(0) value was found to be 20, which is as high as that of soybean lecithin. Glycolipid biosurfactant has a great potential for the formation of W/O microemulsion without using any cosurfactants.

Mannosylerythritol lipid-A (MEL-A) is one of the most promising glycolipid biosurfactants, and abundantly produced by Pseudozyma yeasts. MEL-A gives not only excellent self-assembling properties but also a high binding affinity toward human immunoglobulin G (HIgG). In this study, three kinds of MEL-A were prepared from methyl myristate [MEL-A (m)], olive oil [MEL-A (o)], and soybean oil [MEL-A (s)], and the effect of interfacial properties of each MEL-A monolayer on the binding affinity toward HIgG was investigated using surface plasmon resonance (SPR) and the measurement of surface pressure (pi)-area (A) isotherms. Based on GC-MS analysis, the main fatty acids were C(8) and C(10) acids in all MEL-A, and the content of unsaturated fatty acids was 0% for MEL-A (m), 9.1% for MEL-A (o), 46.3% for MEL-A (s), respectively. Interestingly, the acid content significantly influenced on their binding affinity, and the monolayer of MEL-A (o) gave a higher binding affinity than that of MEL-A (m) and MEL-A (s). Moreover, the mixed MEL-A (o)/ MEL-A (s) monolayer prepared from 1/1 molar ratio, which comprised of 27.8% of unsaturated fatty acids, indicated the highest binding affinity. At the air/water interface, MEL-A (o) monolayer exhibited a phase transition at 13 degrees C from a liquid condensed monolayer to a liquid expanded monolayer, and the area per molecule significantly expanded above 13 degrees C, while the amount of HIgG bound to the liquid expanded monolayer was much higher than that bound to liquid condensed monolayer. The binding affinity of MEL-A toward HIgG is thus likely to closely relate to the monolayer packing density, and may be partly controlled by temperature.

Mannosylerythritol lipids (MELs) are one of the most promising biosurfactants known, because of their multifunctionality and biocompatibility. In order to attain an efficient production of MELs, Pseudozyma parantarctica JCM 11752(T), which is a newly identified strain of the genus, was examined for the productivity of MELs at different culture conditions. The yeast strain showed significant cell growth and production of di-acylated MELs even at 36 degrees C. In contrast, on conventional high-level MEL producers including P. rugulosa, the MEL yield considerably decreased with an increase of the cultivation temperature at over 30 degrees C. On P. parantarctica, soybean oil and sodium nitrate were the best carbon and nitrogen sources, respectively. Under the optimal conditions on a shake-flask culture at 34 degrees C, the amount of di-acylated MELs reached over 100 g/L by intermittent feeding of only soybean oil. Interestingly, the yeast strain produced tri-acylated MELs as well as di-acylated ones when grown on the medium containing higher soybean oil concentrations than 8% (vol/vol). The production of tri-acylated MELs was significantly accelerated at between 34 and 36 degrees C. With 20 % (vol/vol) of soybean oil at 34 degrees C, the yield of tri-acylated MELs reached 22.7 g/L. The extracellular lipase activity considerably depended on the culture temperature, and became the maximum at 34 degrees C; this would bring the accelerated production of tri-acylated MELs. Accordingly, the present strain of P. parantarctica provided high efficiency in MEL production at elevated temperatures compared to conventional MEL producers, and would thus be highly advantageous for the commercial production of the promising biosurfactants.

Mannosylerythritol lipids (MELs), which are glycolipid biosurfactants produced by Pseudozyma yeasts, show not only excellent interfacial properties but also versatile biochemical actions. In the course of MEL production from glucose as the sole carbon source, P. antarctica was found to produce unknown glycolipids more hydrophilic than conventional "di-acylated MELs," which have two fatty acyl esters on the mannose moiety. Based on a detailed characterization, the most hydrophilic one was identified as 4-O-(3'-O-alka(e)noyl-beta-D: -mannopyranosyl)-D: -erythritol namely, "mono-acylated MEL." The mono-acylated MEL reduced the surface tension of water to 33.8 mN/m at a critical micelle concentration (CMC) of 3.6 x 10(-4) M, and its hydrophilic-lipophilic balance was tentatively calculated to be 12.15. The observed CMC was 100-fold higher than that of the MELs hitherto reported. Interestingly, of the yeast strains of the genus Pseudozyma, only P. antarctica and P. parantarctica gave the mono-acylated MEL from glucose, despite a great diversity of di-acylated MEL producers in the genus. These strains produced MELs including the mono-acylated one at a rate of 20-25%. From these results, the new MEL is likely to have great potential for use in oil-in-water-type emulsifiers and washing detergents because of its higher water solubility compared to conventional MELs and will thus contribute to facilitating a broad range of applications for the environmentally advanced surfactants.

Kinetic studies on the interactions between self-assembled monolayers of mannosylerythritol lipids (MELs), which are glycolipid biosurfactants abundantly produced by microorganisms, and various classes of immunoglobulins including human IgG, IgA, and IgM were performed using surface plasmon resonance (SPR). The effect of the MEL structure on the binding behavior of HIgG was examined. Assembled monolayers of MEL-A having two acetyl groups on the headgroup gave a high affinity (K(d)=1.7x10(-6)M) toward HIgG, while those of MEL-B or MEL-C having only one acetyl group at C-6' or C-4' position gave little affinity. Our kinetic analysis revealed that the binding manner of HIgG, HIgA (K(d)=2.4x10(-7)M), and HIgM (K(d)=2.2x10(-7)M) to the assembled monolayers of MEL-A is not the monovalent mode but the bivalent mode, and both the first and second rate association constants (k(a1), k(a2)) increase with an increase in the number of antibody binding sites, while those for dissociation (k(d1), k(d2)) changed little. Moreover, we succeeded in directly observing great amounts of HIgG, HIgA, and HIgM bound to MEL-A monolayers using atomic force microscopy (AFM). Finally, we found that MEL-A assembled monolayer binds toward various IgG derived from mouse, pig, rabbit, horse, goat, rat, and bovine as well as human IgG (HIgG), and the only exception was sheep IgG. These results clearly demonstrate that MEL-A assembled monolayers would be useful as noble affinity ligand system for various immunoglobulins.

A carbohydrate ligand system has been developed which is composed of self-assembled monolayers (SAMs) of mannosylerythritol lipid-A (MEL-A) from Pseudozyma antarctica, serving for human immunoglobulin G and M (HIgG and HIgM). The estimated binding constants from surface plasmon resonance (SPR) measurement were Ka = 9.4 x 10(6) M(-1) for HIgG and 5.4 x 10(6) M(-1) for HIgM, respectively. The binding site was not in the Fc region of immunoglobulin but in the Fab region. Large amounts of HIgG and HIgM bound to MEL-A SAMs were directly observed by atomic force microscopy.

The aqueous-phase behavior of mannosylerythritol lipid A (MEL-A), which is a glycolipid biosurfactant produced from vegetable oils by yeast strains of the genus Pseudozyma, was investigated using polarized optical microscopy, small-angle X-ray scattering (SAXS), and differential scanning calorimetry (DSC). MEL-A was found to self-assemble into a variety of distinctive lyotropic liquid crystals including sponge (L3), bicontinuous cubic (V2), and lamella (Lalpha) phases. On the basis of SAXS measurements, we determined the structure of the liquid crystals. The estimated lattice constant for Lalpha was 3.58 nm. DSC measurement revealed that the phase transition enthalpies from the liquid crystal to the fluid isotropic phase were in the range of 0.22-0.44 kJ/mol. Although the present MEL-A phase diagram closely resembled that obtained from relatively hydrophobic poly(oxyethylene) or fluorinated surfactants, the MEL-A L3 region was spread considerably over a wide temperature range (20-65 degrees C) compared to L3 of those surfactants: this is probably due to the unique structure which is molecularly engineered by microorganisms. In this paper, we clarify the aqueous phase diagram of the natural glycolipid biosurfactant MEL-A, and we suggest that the obtained lyotropic crystals are potentially useful as novel nanostructured biomaterials.

The alpha-anomer form of sulfoquinovosyl-monoacylglycerol with a saturated C18 fatty acid (alpha-SQMG-C(18:0)) is a natural sulfolipid that is a clinically promising antitumor agent. It forms vesicles, micelles or an emulsion in water, depending on several physicochemical conditions. The type of aggregate formed appears to strongly influence the bioactivity level. Thus, we investigated the nature of the aggregates in relation to their bioactivities. The structure of the alpha-SQMG-C(18:0) assembly was greatly affected by the type of additive used in the preparation. Emulsification with ethanol and n-decane might be more effective at inhibiting tumor cell growth than the micelle or vesicle preparations. alpha-SQMG-C(18:0) formed an "emulsion-like-aggregate" in ethanol containing an n-decane concentration in the range of 1.03-103 mM: . These ethanol/n-alkane/alpha-SQMG-C(18:0) aggregates inhibited cell growth in a dose-dependent manner, under optimum conditions (i.e., ethanol containing 103 mM: of n-decane or n-dodecane dispersed in phosphate-buffered saline or culture medium). Based on these data, we discuss the relationship between the molecular action of and antitumor activity by alpha-SQMG-C(18:0).

Sulfoquinovosyldiacyglycerol (SQDG) has a wide range of biological activities that make it an attractive compound for the development of new drugs. Chemically synthesized beta-SQDG-C(18:0) (1,2-di-O-stearoyl-3-O-(6-deoxy-6-sulfo-beta-d-glucopyranosyl)-sn-glycerol), for example, has a potent inhibitory effect on DNA polymerases. We investigated the properties of the vesicle form of beta-SQDG-C(18:0) as the monomer has low solubility in water. The structure of the beta-SQDG-C(18:0) vesicles are highly influenced by NaCl concentration in preparation process. At low NaCl concentrations, the beta-SQDG-C(18:0) vesicles have high surface curvature and form small unilamellar vesicles. Increases in NaCl concentration, resulted in decreased surface curvature and a tendency for beta-SQDG-C(18:0) to form large multilamellar vesicles. The small unilamellar vesicles showed a potent inhibitory effect on DNA polymerase beta, whereas the large multilamellar vesicles had no such effect. We investigated further the relationship between vesicle size and activity by preparing smaller vesicles (262, 99 and 43 nm in diameter) using an extrusion technique. These smaller vesicles had a greater inhibitory effect on DNA polymerase beta activity than non-extruded vesicles. beta-SQDG-C(18:0) vesicles, especially those of small size, were effective in DNA polymerase inhibition and are expected to have high applicability in DNA polymerase study.

Thermodynamically stable vesicle (L(alpha1)) formation from glycolipid biosurfactant sponge phase (L(3)) and its mechanism were investigated using a "natural" biocompatible mannosyl-erythritol lipid-A (MEL-A)/L-alpha-dilauroylphosphatidylcholine (DLPC) mixture by varying the composition. The trapping efficiency for calcein and turbidity measurements clearly indicated the existence of three regions: while the trapping efficiencies of the mixed MEL-A/DLPC assemblies at the compositions with X(DLPC)< or =0.1 or X(DLPC)> or =0.8 were almost zero, the mixed assemblies at the compositions with 0.1 or =0.8 were multilamellar vesicles (L(alpha)) with diameter from 2 to 10 microm. Meanwhile, dynamic light scattering (DLS) measurement revealed that the average size of the vesicles at the composition of X(DLPC)=0.3 was 633.2 nm, which is remarkably small compared to other compositions. Moreover, the mixed vesicle solution at the composition of X(DLPC)=0.3 was slightly bluish and turbid and kept its dispersion stability at 25 degrees C for more than 3 months, indicating the formation of a thermodynamically stable vesicle (L(alpha1)). These results exhibited the formation of a thermodynamically stable vesicle (L(alpha1)) with a high dispersibility from the MEL-A/DLPC mixture. The asymmetric distribution of MEL-A and DLPC in the two vesicle monolayers caused by the difference in geometrical structures is very likely to have changed their self-assembled structure from a sponge phase (L(3)) to a thermodynamically stable vesicle (L(alpha1)).

Physicochemical properties of 1,2-di-O-stearoyl-3-O-(6-deoxy-6-sulfo-alpha-D-glucopyranosyl)-sn-glycerol (alpha-SQDG-C(18:0)) and 1,2-di-O-stearoyl-3-O-(6-deoxy-6-sulfo-beta-D-glucopyranosyl)-sn-glycerol (beta-SQDG-C(18:0)) in monolayer and bilayer membranes were examined. Surface pressure measurements in monolayer membranes indicated the molecular area of beta-SQDG-C(18:0) to be slightly smaller than that of alpha-SQDG-C(18:0). In bilayer membranes, the phase transition temperature and the enthalpy of beta-SQDG-C(18:0) were higher than those of alpha-SQDG-C(18:0), while the trapping efficiency of beta-SQDG-C(18:0) vesicles was lower. The results suggested tighter packing with beta-SQDG-C(18:0) than alpha-SQDG-C(18:0), due to differences in the head group stereochemistry. High-performance liquid chromatography-electrospray ionization ion trap mass spectrometry (HPLC-ESI-MS) data and computational modeling studies provided supporting evidence for morphological differences. In both monolayer and bilayer membranes, the affinity of beta-SQDG-C(18:0) with cholesterol was greater than that of alpha-SQDG-C(18:0), again due to the differences in head group properties. Turbidity measurement and microscopic examination of alpha- and beta-SQDG-C(18:0)/cholesterol mixtures confirmed formation of large vesicles. The addition of cholesterol to SQDG-C(18:0) optimized membrane formation and stabilized its structure.

The article presents a study on the improvement of celloboise lipids (CL) production using the strains of Cryptococcus humicola (Cr. humicola) and their physiochemical properties. The study used thin-layer chromatography (TLC) to analyze glycolipids from the strain JCM 1461 and nuclear magnetic resonance (NMR). Results showed that the strain JCM 1461 has produced 16-O-(2",3"4",6'-tetra-O-acetyl-β-cellobiosyl)-2-hydroxy-hexadecanoic acid with asymmetric bolaform glycolipid property.

Abstract: The photooxidative mineralizations of two microorganisms-produced glycolipid biosurfactants 4-O-(4′,6′-di-O-acetyl-2′,3′-di-O-alkanoyl-β-d-mannopyranosyl)-d-erythritol (MEL-A) and 1-O-(6′-O-acetyl-2′,3′-di-O-alkanoyl-β-d-mannopyranosyl)-d-erythritol (MEL-B) were examined by monitoring the temporal changes in UV absorption, the time profiles of CO2 evolution and the changes in interfacial tension occurring by an advanced oxidation process in the presence of a metal-oxide (TiO2). Features of their mineralization are compared to the photomineralization of the anionic dodecylbenzene sulfonate (DBS) surfactant carried out under otherwise identical conditions. The adsorption of surfactants on the TiO2 surface and the positions of attack of the photogenerated ls on the surfactants’ structure were assessed by molecular orbital (MO) calculations of partial charges and frontier orbital electron densities, respectively. The photodegradation of DBS was faster than the MELs as also evidenced by surface tension measurements, whereas the photomineralization of the anionic DBS surfactant was definitely slower than that of the MEL biosurfactants due to the hydrophobic alkyl chain in the DBS structure. Possible initial mechanistic stages of the photooxidation of MEL-A and MEL-B are proposed based on experimental data and comparison with MO calculations. [Copyright &y& Elsevier]