Antibacterial Activities of Sophorolipids and Nisin and Their Combination against Foodborne Pathogen Staphylococcus aureus

In food processing, both preservatives and surfactants are usually added to ensure the quality of food. Sophorolipids (SLs) are extracellular biosurfactants produced by yeasts that have antibacterial activities. Nisin is a polycyclic antibacterial peptide produced by Lactococcus lactis. In this study, the possibility of using SLs alone as a preservative and in combination with nisin as a compound additive is investigated. SLs and nisin exhibit effective antibacterial activities against foodborne pathogen Staphylococcus aureus with minimum inhibitory concentrations of 32 and 0.5 µg mL^?1, respectively. Both SLs and nisin increase cell wall and cell membrane permeability of S. aureus, enhance intracellular content release and activities of extracellular ALP and β‐galactosidase, and reduce respiratory‐chain dehydrogenase activity of the pathogen. Fractional inhibitory concentration (FIC) index for SLs and nisin combination is 1.5, indicating the effect of SLs with nisin is additive. Results of the study indicate that SLs could be used alone as a preservative or in combination with nisin as a promising compound additive with functions of both emulsifier and preservative. Practical Applications: SLs could be used alone as a preservative in food industry or in combination with nisin as a promising compound additive with functions of both emulsifier and preservative.     

The Yeast ATF1 Acetyltransferase Efficiently Acetylates Insect Pheromone Alcohols: Implications for the Biological Production of Moth Pheromones

Many moth pheromones are composed of mixtures of acetates of long‐chain (≥10 carbon) fatty alcohols. Moth pheromone precursors such as fatty acids and fatty alcohols can be produced in yeast by the heterologous expression of genes involved in insect pheromone production. Acetyltransferases that subsequently catalyze the formation of acetates by transfer of the acetate unit from acetyl‐CoA to a fatty alcohol have been postulated in pheromone biosynthesis. However, so far no fatty alcohol acetyltransferases responsible for the production of straight chain alkyl acetate pheromone components in insects have been identified. In search for a non‐insect acetyltransferase alternative, we expressed a plant‐derived diacylglycerol acetyltransferase (EaDAcT) (EC 2.3.1.20) cloned from the seed of the burning bush (Euonymus alatus) in a yeast system. EaDAcT transformed various fatty alcohol insect pheromone precursors into acetates but we also found high background acetylation activities. Only one enzyme in yeast was shown to be responsible for the majority of that background activity, the acetyltransferase ATF1 (EC 2.3.1.84). We further investigated the usefulness of ATF1 for the conversion of moth pheromone alcohols into acetates in comparison with EaDAcT. Overexpression of ATF1 revealed that it was capable of acetylating these fatty alcohols with chain lengths from 10 to 18 carbons with up to 27‐ and 10‐fold higher in vivo and in vitro efficiency, respectively, compared to EaDAcT. The ATF1 enzyme thus has the potential to serve as the missing enzyme in the reconstruction of the biosynthetic pathway of insect acetate pheromones from precursor fatty acids in yeast.     

Behenic acid esters: kinetics and properties

Esterification of commercial behenic acid with fatty alcohols (decanol, lauryl alcohol, myristyl alcohol and cetyl alcohol) was studied using n‐butyl benzene as solvent and tetra n‐butyl titanate (TBT) as catalyst. The effects of reaction temperature (165–185 °C) and catalyst loading (0.3–1.2%, wt/wt) on the conversion of behenic acid were studied for all the alcohols. The reaction showed first order dependence with respect to fatty alcohols and behenic acid concentration. The values of activation energy obtained for the catalyzed reaction of behenic acid with decanol, lauryl alcohol, myristyl alcohol and cetyl alcohol were found to be 68.1, 69.2, 64.5 and 67.3 kJ mol^?1, respectively. The values of activation energy obtained for the uncatalyzed reaction of behenic acid with decanol, lauryl alcohol, myristyl alcohol and cetyl alcohol were found to be 86.2, 79.6, 78.6 and 87.1 kJ mol^?1, respectively. Physical properties such as melting point, viscosity, specific gravity, and refractive index of these esters are reported.     

Synthesis of fatty acid esters from acid oils using lipase B from Candida antarctica

Esterification of corn and sunflower acid oils with straight‐ and branched‐chain alcohols were conducted using lipase B from Candida antarctica (Novozym 435) in n‐hexane. Sunflower acid oil consisted of 55.6% free fatty acids and 24.7% triacylglycerols, while the free fatty acids and triacylglycerols contents of corn acid oil were 75.3% and 8.6%, respectively. After 1.5 h of methanolysis of sunflower acid oil, the highest fatty acid methyl ester content (63.6%) was obtained at 40 °C and the total fatty acid/methanol molar ratio was 1/1, using 15% enzyme based on acid oil weight. The conversion of both acid oils with straight‐ and branched‐chain alcohols was not significantly affected by the chain length of the alcohols. However, the lowest fatty acid methyl ester content (50%) was obtained in the reaction of corn acid oil with methanol. Sunflower acid oil was converted to fatty acid esters using primer alcohols such as n‐propanol, i‐ and n‐butanol, n‐amylalcohols, n‐octanol, and a mixture of amylalcohol isomers, resulting in a fatty acid ester content of about 70% at 40 °C.     

Effect of Alcohol Addition to Gasoline on Soot Distribution Characteristics in Laminar Diffusion Flames

Alcohols are widely used as alternative fuel for spark ignition engines to reduce soot emission. 2D soot distributions of alcohols‐gasoline blends were studied in laminar diffusion flames by two‐color laser‐induced incandescence technique. The soot volume fraction decreases significantly with the alcohol blending ratio. At the same blending ratio, the soot‐reducing ability declined with the carbon length of the alcohol, but the reducing effect for short‐chain alcohols, i.e., methanol and ethanol, deteriorated at high blending ratio while it remained constant for the long‐chain alcohol n‐butanol. At fixed oxygen content, the soot‐reducing ability of a short‐chain alcohol is still higher than that of a long‐chain alcohol, i.e., not only the oxygen content but also the molecular structure dominates the soot‐reducing ability.     

Kinetic Analysis of Terminal and Unactivated C?H Bond Oxyfunctionalization in Fatty Acid Methyl Esters by Monooxygenase‐Based Whole‐Cell Biocatalysis

The alkane monooxygenase AlkBGT from Pseudomonas putida GPo1 constitutes a versatile enzyme system for the ω‐oxyfunctionalization of medium chain‐length alkanes. In this study, recombinant Escherichia coli W3110 expressing alkBGT was investigated as whole‐cell catalyst for the regioselective biooxidation of fatty acid methyl esters to terminal alcohols. The ω‐functionalized products are of general economic interest, serving as building blocks for polymer synthesis. The whole‐cell catalysts proved to functionalize fatty acid methyl esters with a medium length alkyl chain specifically at the ω‐position. The highest specific hydroxylation activity of 104 U g_CDW⁻¹ was obtained with nonanoic acid methyl ester as substrate using resting cells of E. coli W3110 (pBT10). In an optimized set‐up, maximal 9‐hydroxynonanoic acid methyl ester yields of 95% were achieved. For this specific substrate, apparent whole‐cell kinetic parameters were determined with a V_max of 204±9 U g_CDW⁻¹, a substrate uptake constant (K_S) of 142±17 μM, and a specificity constant V_max/K_S of 1.4 U g_CDW⁻¹ μM ⁻¹ for the formation of the terminal alcohol. The same E. coli strain carrying additional alk genes showed a different substrate selectivity. A comparison of biocatalysis with whole cells and enriched enzyme preparations showed that both substrate availability and enzyme specificity control the efficiency of the whole‐cell bioconversion of the longer and more hydrophobic substrate dodecanoic acid methyl ester. The efficient coupling of redox cofactor oxidation and product formation, as determined in vitro, combined with the high in vivo activities make E. coli W3110 (pBT10) a promising biocatalyst for the preparative synthesis of terminally functionalized fatty acid methyl esters.     

Lipophilic Extractives in Eucalyptus globulus Kraft Pulps. Behavior during ECF Bleaching

Abstract

The composition of E. globulus kraft pulp lipophilic extractives and their behaviour during an ECF (DEDED) bleaching sequence were investigated. Sterols; fatty acids, including several α‐ and ω‐hydroxyfatty acids; and long‐chain aliphatic alcohols are the major lipophilic extractives of the unbleached pulp. During the bleaching, about 80% of the aliphatic extractives are removed from pulp (ca. 70% of the sterols, 70% of the fatty acids, and 90% of the long‐chain aliphatic alcohols). The decrease of sterols is mainly due to the degradation of β‐sitosterol by chlorine dioxide, while the decrease of fatty acids and alcohols is essentially assigned to their extraction and elimination with the alkaline filtrates. The major chemical transformations in pulp extractives composition and structure occur in the last bleaching stages.     

Microbial Conversion of Vegetable Oil to Rare Unsaturated Fatty Acids and Fatty Alcohols by an <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis> Isolate

We isolated two new microorganisms capable of converting vegetable oil to several rare unsaturated fatty acids and rare unsaturated fatty alcohols from a soil sample. The strains were identified as belonging to the same genus and species, Aeromonas hydrophila. The rare unsaturated fatty acids and rare unsaturated fatty alcohols were accumulated as a wax ester form by the strains. Compared to other strains, the A. hydrophila isolates effectively decreased fatty acid chain lengths and converted rapeseed oil, which is rich in 9-C18:1 fatty acid, into rare fatty acids, such as 7-C16:1 fatty acid and 5-C14:1 fatty acid. Furthermore, the A. hydrophila isolates converted the resulting fatty acids to rare unsaturated fatty alcohols, such as 7-C16:1 fatty alcohol and 5-C14:1 fatty alcohol. The isolates also converted safflower oil, which is rich in 9,12-C18:2 fatty acid, to 7,10-C16:2 fatty acid, 5,8-C14:2 fatty acid, 9,12-C18:2 fatty alcohol, 7,10-C16:2 fatty alcohol, and 5,8-C14:2 fatty alcohol. 7,10,13-C16:3 fatty acid, 9,12,15-C18:3 fatty alcohol, and 7,10,13-C16:3 fatty alcohol were also converted from linseed oil, which is rich in 9,12,15-C18:3 fatty acid, by the A. hydrophila isolates. These fatty acids and fatty alcohols are rarely found in natural oils. Since decreasing fatty acid carbon chain lengths from the carboxyl end and reducing unsaturated fatty acids to unsaturated fatty alcohols are both difficult reactions to accomplish by chemical means, we suggest that these A. hydrophila isolates may facilitate introduction of new bioprocess for producing rare unsaturated fatty acids and rare unsaturated fatty alcohols, especially fatty alcohols harboring more than two double bonds.     

Esterification of Fatty Acid by Zirconic Catalysts

Sulfated zirconia (SO₄ ^2?/ZrO₂) was proved as an efficient catalyst for the esterification of fatty acids and alcohols, the alkyl esters of long chain fatty acids are wildly used in industry and daily life. In this paper, the esterification of long chain fatty acid (both saturated and unsaturated) with simple alcohol catalyzed by SZ catalyst was presented. It is found that the degree of esterification reaction of the fatty acids with 1-butanol has a close relationship with the length and unsaturation degree of the chain. The longer the carbon chain is, the more difficult it is to react; the higher the unsaturation level is, the more difficult it is to respond. The related regulation will be confirmed and serve as the dictator for other reactions within this system.     

Jatropha oil and karanja oil as carbon sources for production of sophorolipids

Biosurfactants like sophorolipids (SL) are mild and environmentally friendly surfactants to be used in cosmetics and health care products. In addition to surfactant properties, SL also possess antimicrobial and skin healing properties. SL are produced by microbial fermentation using refined vegetable oils with glucose as a carbon source. This affects the economics of the production of SL. In the present work, non‐traditional oils like jatropha oil, karanja oil, and neem oil were used as newer feedstock for fermentative production of SL using Starmerella bombicola (ATCC 22214). In the fermentation, jatropha oil and karanja oil gave 6.0 and 7.6 g/L of SL (mainly lactonic form), respectively. HPLC, liquid chromatography–mass spectrometer, and ¹H NMR of crude SL obtained from fermentation broth showed lactonic form of two major SL. Oleic acid and linoleic acid were preferentially consumed over other fatty acids by the organism. Neem oil gave lower yield, i.e., 2.63 g/L of SL (mainly acidic form). Practical applications: Jatropha oil and karanja oil are one of the non‐traditional oils grown wildly in India that have large potential that is still to be explored. These oils contain non‐glycerides components that exclude their use as edible oil. These oils can be used as substrate for SL that can find novel applications like in soil remediation, skin care production, antimicrobial agents, low foaming detergents, and food additives. The current study has provided proof of concept work that has indicated the potential of these oils to be used as substrate for SL. It has opened new avenues and there is further scope to improve the yield by validating the process parameters like aeration rate, residual substrate recycle and pH control.     

Lipophilic components in black currant seed and pomace extracts

The nature of the fatty acids and other lipophilic components in extracts from black currant seed and pomace (containing seed) were investigated, with a view to highlighting any potential uses. The same non‐hydroxylated fatty acids were the major components in both types of extract, but total levels were less in pomace (75 582 mg 100 g^?1 oil) than in seed alone (90 972 mg 100 g^?1 oil) and there were less unsaturated fatty acids, including GLA (8653 and 12 625 mg 100 g^?1 oil, respectively), but long chain n‐20:0 – n‐30:0 fatty acids (4080 and 437 mg 100 g^?1 oil, respectively) were greatly increased in pomace. Phytosterols (mainly β‐sitosterol), saturated n‐20:0 – n‐30:0 policosanols, ω‐hydroxy fatty acids (mainly 16‐hydroxy 16:0) and 2‐hydroxy fatty acids (mainly 2‐hydroxy 24:0) were present at much greater levels in pomace (2496, 2097, 958 and 46 mg 100 g^?1 oil, respectively) than in seed (553, 108, 161, and 1 mg 100 g^?1 oil, respectively). The pomace extract is a useful source of fatty acids, phytosterols and policosanols with potential functional properties. Practical applications: The study investigated the lipophilic components in isohexane extracts from black currant seed and pomace (containing seed). Only pomace extracts had substantial amounts of phytosterols and policosanols that have potential as cholesterol‐lowering agents, whereas fatty acids such as GLA, that has anti‐inflammatory properties, are mainly in the seed.     

Effects of Rice Bran Oil Enriched with n-3 PUFA on Liver and Serum Lipids in Rats

Lipase-catalyzed interesterification was used to prepare different structured lipids (SL) from rice bran oil (RBO) by replacing some of the fatty acids with α-linolenic acid (ALA) from linseed oil (LSO) and n-3 long chain polyunsaturated fatty acids (PUFA) from cod liver oil (CLO). In one SL, the ALA content was 20% whereas in another the long chain n-3 PUFA content was 10%. Most of the n-3 PUFA were incorporated into the sn-1 and sn-3 positions of triacylglycerol. The influence of SL with RBO rich in ALA and EPA + DHA was studied on various lipid parameters in experimental animals. Rats fed RBO showed a decrease in total serum cholesterol by 10% when compared to groundnut oil (GNO). Similarly structured lipids with CLO and LSO significantly decreased total serum cholesterol by 19 and 22% respectively compared to rice bran oil. The serum TAGs level of rats fed SLs and blended oils were also significantly decreased by 14 and 17% respectively compared to RBO. Feeding of an n-3 PUFA rich diet resulted in the accumulation of long chain n-3 PUFA in various tissues and a reduction in the long chain n-6 PUFA. These studies indicate that the incorporation of ALA and EPA + DHA into RBO can offer health benefits.     

Aldehyde‐Catalyzed Transition Metal‐Free Dehydrative β‐Alkylation of Methyl Carbinols with Alcohols

Different to the borrowing hydrogen strategy in which alcohols were activated by transition metal‐catalyzed anaerobic dehydrogenation, the direct addition of aldehydes was found to be an effective but simpler way of alcohol activation that can lead to efficient and green aldehyde‐catalyzed transition metal‐free dehydrative C‐alkylation of methyl carbinols with alcohols. Mechanistic studies revealed that the reaction proceeds via in situ formation of ketones by Oppenauer oxidation of the methyl carbinols by external aldehydes, aldol condensation, and Meerwein–Ponndorf–Verley (MPV)‐type reduction of α,β‐unsatutated ketones by substrate alcohols, affording the useful long chain alcohols and generating aldehydes and ketones as the by‐products that will be recovered in the next condensation to finish the catalytic cycle.     

Neue Tenside aus gemischten Guerbet-Alkoholen

New Surfactants from Mixed Guerbet-Alcohols 2-Benzyl fatty alcohols can be prepared in good yields by mixed Guerbet-reaction of fatty alcohols in the range of chain lengths from C₈–C₁₄ with benzyl alcohol. The polyglycolethers and polyglycolethersulfates from these mixed Guerbet-alcohols are very effective new surfactants with a better biodegradibility as compared to nonylphenol polyglycolethers.     

Distinctive Lipid Composition of the Copepod Limnocalanus macrurus with a High Abundance of Polyunsaturated Fatty Acids

We studied the copepod Limnocalanus macrurus for seasonal variation in the composition of fatty acids, wax esters and sterols in large boreal lakes, where it occurs as a glacial‐relict. Vast wax ester reserves of Limnocalanus were accumulated in a period of only two months, and comprised mono‐ and polyunsaturated fatty acids (PUFA) and saturated fatty alcohols. In winter, the mobilization of wax esters was selective, and the proportion of long‐chain polyunsaturated wax esters declined first. PUFA accounted for >50 % of all fatty acids throughout the year reaching up to ca. 65 % during late summer and fall. Long‐chain PUFA 20:5n‐3 and 22:6n‐3 together comprised 17–40 % of all fatty acids. The rarely reported C24 and C26 very‐long‐chain PUFA (VLC‐PUFA) comprised 6.2 ± 3.4 % of all fatty acids in August and 2.1 ± 1.7 % in September. The VLC‐PUFA are presumably synthesized by Limnocalanus from shorter chain‐length precursors because they were not found in the potential food sources. We hypothesize that these VLC‐PUFA help Limnocalanus to maximize lipid reserves when food is abundant. Sterol content of Limnocalanus, consisting ca. 90 % of cholesterol, did not show great seasonal variation. As a lipid‐rich copepod with high abundance of PUFA, Limnocalanus is excellent quality food for fish. The VLC‐PUFA were also detected in planktivorous fish, suggesting that these compounds can be used as a trophic marker indicating feeding on Limnocalanus.     

Conformational behaviors of syndiotactic polystyrene in chloroform revealed by intrinsic fluorescence technique

Conformational behaviors of syndiotactic polystyrene (sPS) in chloroform were investigated by detecting its concentration dependence of intrinsic fluorescence in the range from 25 to −62 °C. For the dimer emission, the coexistence of T-shaped configuration with sandwich configuration was observed and the contribution of sandwich configuration increases with increasing sPS concentration in the system. In addition, three conformations are observed in sPS/CHCl₃ systems: (I) coil phase; (II) coil-to-helix transformation; (III) helix-to-network transformation. The conformational ordering of polymer chain in super dilute sPS/CHCl₃ systems during isothermal process indicated that micro-gel formed when the concentration was higher than 0.2 g L⁻¹, and single helix formed at 0.09 ∼ 0.2 g L⁻¹ proceeds through a unimolecular or self-organization mechanism and no helical aggregates were found. The helix was formed by clustering of several chain segments when the polymer concentration was below 0.09 g L⁻¹. Moreover, a model was proposed to describe the elasticity of sPS/CHCl₃ systems near and beyond the critical percolation concentration.     

Chemo‐enzymatic synthesis of lipophilic ferulates and their evaluation for antioxidant and antimicrobial activities

An efficient chemo‐enzymatic synthesis of ferulic acid‐based structured lipids mimicking triacylglycerol with a pendant phenolic moiety was carried out for the first time. Initially, ferulic acid was reduced to coniferyl alcohol, followed by its esterification with fatty acids. The key step in the synthesis was dihydroxylation of the olefinic side chain of coniferyl ester, which was eventually esterified with fatty acids to generate phenolic structured lipids. Two such compounds of varying fatty acid chain lengths were synthesized in good yield. Structural confirmation of both compounds is based on IR, ¹H and ¹³C NMR, and MS techniques. The synthesized compounds were tested for in vitro antioxidant and antimicrobial activities. Both compounds exhibited moderate to good antioxidant activity. The phenolic structured lipid with only shorter‐chain fatty acids showed antibacterial activity. Both compounds did not show any antifungal activity.     

Fatty alcohols

“Fatty” or higher alcohols are mostly C₁₁ to C₂₀ monohydric compounds. In probably no other homologous aliphatic series is the current balance between natural and synthetic products so vividly evident. Natural sources, such as plant or animal esters (waxes), can be made to yield straight chain (normal) alcohols with a terminal (primary) hydroxyl, along with varying degrees of unsaturation. In the past, socalled fatty alcohols were prepared commercially by three general processes from fatty acids or methyl esters, occasionally triglycerides. Fatty acids add hydrogen in the carboxyl group to form fatty alcohols when treated with hydrogen under high pressure and suitable metal catalysts. By a similar reaction, fatty alcohols are prepared by the hydrogenation of glycerides or methyl esters. Fatty alcohols are also prepared by the sodium reduction of esters of fatty acids in a lower molecular weight alcohol. The sodium reduction method was ordinarily too expensive; it was displaced early by the other methods; finally most unsaturated alcohols made by this route were largely replaced. Methyl ester reduction continues to provide perhaps 20% of the saturated fatty alcohols, and selective hydrogenation with the use of special catalysts such as copper or cadmium oxides was developed for the production of oleyl alcohol. Synthetic or petroleum technology for long chain alcohols include the Ziegler process, useful for straight chain, even-numbered saturated products. A second is the carbonylation and reduction of olefins affording medium or highly branched chain alcohols. Paraffin oxidation affords mixed primary alcohols. Fatty alcohols undergo the usual reactions of alcohols. They may be reacted with ethylene oxide to yield a series of polymeric polyoxyethylene alcohols or with acetylene under pressure to yield vinyl ethers or with vinyl acetate to give vinyl ethers.     

An Enantioconvergent Synthesis of (R)‐4‐Aryloxy‐1‐butyne‐3‐ols for Prostanoid Side Chains

The single enantiomer title alcohols, useful as ω‐side chain precursors for pharmaceutically important prostaglandin analogues were synthesised from the corresponding racemic alcohols by a convenient 4‐step sequence. After enzymatic acylation of the alcohol with a vinyl carboxylate, the residual (S)‐alcohol in the mixture was converted to the mesylate. Subsequent displacement with the corresponding carboxylate anion, followed by enzymatic deacylation gave the desired (R)‐alcohol. In this way, all of the starting alcohol was utilised without the need for separation of the starting material and product after the bioresolution.     

Free Fatty Acids Reduction in Waste Cooking Oil by Rhodosporidium toruloides and Simultaneous Carotenoids,Lipids, and PAL Enzyme Production in a Two-Phase Culture System

Waste cooking oil (WCO) is a problematic waste product that contains free fatty acids (FFAs), preventing it from being valorized easily as biodiesel and poses an environmental hazard if incorrectly disposed. The use of WCO as a carbon source for Rhodosporidium toruloides (R. toruloides) using a two-phase culture system is developed. The normal growth of R. toruloides when cultured in WCO (OD₆₀₀ 52) reveals its ability to use a hydrophobic substrate as the carbon source compared to glucose (OD₆₀₀ 51.9). Interestingly, the extracellular lipase activity when R. toruloides is grown on WCO is 14.4 U mL⁻¹ compared to when grown on glucose (2.4 U mL⁻¹). Additionally, FFA levels in WCO are reduced from 2% to 0.2% at end of fermentation, suggesting that R. toruloides can consume FFA. Furthermore, higher yield of beneficial products: β-carotene (4.57 µg mL⁻¹), torularhodin (4.2 µg mL⁻¹), fatty acids (1 mg mL⁻¹), and phenylalanine ammonia-lyase (PAL) enzyme (0.12 µmol mg⁻¹) are produced when WCO is the carbon source, compared to glucose (4.1 µg mL⁻¹ β-carotene, 3.0 µg mL⁻¹ torularhodin, 1 mg mL⁻¹ of fatty acids, and 0.096 µmol mg⁻¹ PAL enzyme). This is a first study that shows R. toruloides can grow on hydrophobic carbon source.