Optimization of production of docosahexaenoic acid (DHA) byThraustochytrium aureum ATCC 34304

By varying culture carbon source, lipid content in mycelium ofThraustochytrium aureum ATCC 34304 varied widely in the range 1–25% of biomass weight. Docosahexaenoic acid (DHA) content of mycelium lipid was higher (65–76%) when biomass lipid content was very low (1–2%) and lower (40–50%) when biomass contained a high lipid content (14–18%). DHA yields from glucose, starch and maltose were 270, 325 and 334 mg/L, respectively. DHA yield and content of biomass was optimal at an initial culture pH of 6.0. During the culture cycle ofT. aureum, DHA content in lipids remained relatively constant with optimal DHA yield being observed after six days. Biomass, lipid content in biomass, DHA content in biomass and DHA yield were all optimal at a cultivation temperature of 28°C. However, the proportion of DHA in lipids declined with increase in temperature. Biomass, lipids in biomass and DHA yields were 13%, 42% and 47% higher, respectively, din light-exposed cultures as compared to dark cultures. A maximum yield of DHA of 511 mg/L was observed in light- exposed cultures containing 2.5% starch, where lipids accounted for over 20% of biomass dry weight. To whom correspondence should be addressed.     

A physiological study on growth and dibenzothiophene (DBT) desulfurization characteristics of Gordonia sp. CYKS1

Physiological characteristics of DBT desulfurization and cell growth ofGordonia sp. CYKS1 were investigated. It exhibited a preference to ethanol in a medium containing two carbon sources, ethanol and one of the carbohydrates used, glucose, sucrose, maltose, and galactose although it consumed both carbon sources simultaneously. Cell growth on ethanol or glucose followed the Monod kinetics. The optimal range of pH for the desulfurization of DBT and the cell growth was 7 to 8. The desulfurization rate decreased about 30% at pH 6, and no significant desulfurization or cell growth was observed at pH 5. As the initial DBT concentration increased up to 1.5 mM, the desulfurization rate also increased while no significant changes in the growth rate were observed. The maximum desulfurization rate was 12.50 μmol L^-1 h^-1 at an initial DBT concentration of 1.5 mM. Cell growth and desulfurization activity were severely inhibited by the presence of 2-hydroxybiphenyl (2-HBP). When 0.05 mM of 2-HBP was added at the beginning, both of the desulfurization rate and cell growth rate decreased about 20%. It was found that cell growth and desulfurization were completely inhibited in the presence of 2-HBP at 0.15 mM or a higher concentration. The inhibition by 2,2′-dihydroxybiphenyl (DHBP) was less severe than 2-HBP. About 80% of desulfurization activity was retained in the presence of 2,2′-DHBP at 0.4 mM.     

Mechanism by which noninhibitory concentrations of glucose increase inhibitory activity ofp-coumaric acid on morning-glory seedling biomass accumulation

Noninhibitory levels of glucose-C [ 72 µg carbon (C)/g soil] increased the inhibitory activity ofp-coumaric acid on morning-glory seedling biomass accumulation in Cecil B_t-horizon soil. The amount ofp-coumaric acid required for a given level of inhibition of shoot and seedling biomass accumulation decreased as the concentration of glucose increased. Soil extractions with neutral EDTA (0.25 M, pH 7) after addition of combinations ofp-coumaric acid and glucose (concentrations ranging from 0 to 1.25 µmol/g soil) to the soil showed that utilization ofp-coumaric acid by microbes decreased linearly as the concentration of glucose increased. The increased inhibitory activity of a given concentration ofp-coumaric acid in the presence of glucose was not due to a reduction in soil sorption ofp-coumaric acid or effects of nitrogen-limited microbial growth. Noninhibitory levels of phenylalanine andp-hydroxybenzoic acid slowed the utilization ofp-coumaric acid by microbes in a similar manner as glucose. The presence of methionine, however, did not affect the rate ofp-coumaric acid utilization by microbes. These observations suggest that differential utilization of individual molecules in organic mixtures by soil microbes can modify, and in this case increase, the effectiveness of a given concentration of an inhibitor such asp-coumaric acid on the inhibition of seedling growth such as morning-glory.The use of trade names in this publication does not imply endorsement by the United States government or the North Carolina Agricultural Research Service of products named, nor criticism of similar ones not mentioned.     

Biodegradation of toluene and dimethyl sulfide in a cocultured biofilter

Biofiltration was performed for 50 days in a perlite-packed biofilter (8 cm I.D.x 105 cm height) for the simultaneous removal of toluene and dimethyl sulfide (DMS). Two strains,Rhodococcus pyridinovorans PYJ-1 andGordonia sihwaniensis PKL-1, were cocultured in the biofilter. Removal efficiencies of toluene and DMS at an empty bed residence time (EBRT) of 3 min were 80–85% and 40–45%, respectively, for an input concentration of 2.5–3.0 mg/L of toluene and 1.5–2.0 mg/L of DMS. The pH of the perlite column was maintained at 7.0–7.2, and the moisture content varied from 61% at the bottom to 51% at the top. Starting from the same initial cell concentration (2.4 x 10₆ CFU/g of wet packing) the number ofR. pyridinovorans PYJ-1 was 2.5 times higher compared with that of G.sihwaniensis PKL-1 after 50 days of operation.     

Studies on the preparation and dental properties of antibacterial polymeric dental restorative composites containing alkylated ammonium chloride derivatives

The preparation of antibacterial polymeric dental restorative composites (PDRC) was carried out using various alkylated ammonium chloride derivatives as the antibacterial compound. To improve the mechanical properties of PDRC, the surface of the silica filler was hydrophobically treated with γ-methacryloxypropyltrimethoxysilane. The diametral tensile strength, flexural strength, and Knoop hardness of PDRC were measured and the antibacterial effect was also investigated. The addition of alkylated ammonium chloride derivatives to the bisphenol-A glycerolate dimethacrylate/triethyleneglycol dimethacrylate resin matrix enhanced the antibacterial properties of PDRC. It was also discovered that alkylated ammonium chloride derivatives with a higher chain length between the ammonium and the acryl (or methacryl) functional groups reduced the mechanical properties of PDRC. In addition, it was found that the optimum concentration of the antibacterial compound was about 3 wt% based upon resin matrix, and the optimum chain length between functional groups of alkylated ammonium chloride derivatives was 2–3 carbons.     

Variation in fatty acid content and seed weight in some lauric acid richCuphea species

Significant genetic variation for lauric acid (12∶0) and capric acid (10∶0) composition and seed weight was measured within lauric acid-rich, self-pollinating germplasm accessions ofCuphea wrightii, C. tolucana, andC. lutea. Means and ranges of individual plant progenies for 12∶0 content ofC. wrightii accessions was 60.5±.63% (49.8±65.8%), 10∶0 content was 23.7±.54% (18.6±33.0%), and 1000-seed weight was 1.50±.03 g (1.20–2.47 g). Progenies of single plant selections carried to the S₂ generation exhibited reduced variability within selections, but significant variation among selections for 12∶0, 10∶0 and 1000-seed weight. Variation among single plant selections ofC. tolucana was less than that ofC. wrightii and attributed to a restricted germplasm base. Means and ranges for 12∶0 content were 61.6±.47% (59.2–69.9%), 10∶0 was 22.3±.62% (11.7–25.3%), and 1000-seed weight was 1.40±.05 g (0.90–1.69 g).Cuphea lutea has a significantly different 12∶0−10∶0 profile than the other lauric acid-rich species. Means and ranges for 12∶0 were 36.8±.14% (33.7–40.8%), 10∶0 was 21.8±.08% (16.4–23.9%), 1000-seed weight was 2.26±.02 g (1.82–272 g). The 1000-seed weight was highly positively correlated with 8∶0, 10∶0, 18∶1 and 18∶2 contents and highly negatively correlated with 12∶0, 14∶0 and 16∶0 in bothC. wrightii andC. tolucana. No such relationship was found forC. lutea. A highly significant negative correlation was also measured for 12∶0 and 10∶0 contents inC. wrightii andC. tolucana.     

Effect of phenol on Β-carotene content in total carotenoids production in cultivation of<Emphasis Type="Italic">Rhodotorula glutinis</Emphasis>

The composition of carotenoids produced byR. glutinis was observed to be dependent upon the addition of phenol into medium. A stimulatory effect of phenol on Β-carotene ofRhodotorula glutins K-501 grown on glucose was investigated. Carotenoids produced byRhodotorula glutinis K-501 were identified to torularhodin, torulene and Β-carotene, whose composition was 79.5%, 6.4% and 14.1%, respectively. The Β-carotene content increased up to 35% when phenol was added to culture media at 500 ppm. The ratio of torularhodin decreased with increasing phenol concentration, while torulene content was almost constant.     

Effect of temperature and chainlength on the densities and molal volumes ofn-alkyl chlorides

The densities ofn-alkyl chlorides from pentyl chloride to hexadecyl chloride were determined at temperatures between 15–80°C at 5°C intervals. The densities increase linearly with temperature and chainlength. A four-constant equation, V=n/(−3.6640 × 10⁻⁵ T+0.07151)+1/(−5.6526 × 10⁻⁵ T+0.04243), was formulated. This formula accurately predicted the molal volume and, hence, the density for all then-alkyl chlorides at any temperature within the range.     

Purification and characterization of thermophilic and alkalophilic tributyrin esterase fromBacillus strain A30-1 (ATCC 53841)

An extracellular esterase (EC 3.1.1.1) from a thermophilicBacillus A30-1 (ATCC 53841) was purified 139-fold to homogeneity by sodium chloride (6 M) treatment, ammonium sulfate fractionation (30–80%) and phenyl-Sepharose CL-6B column chromatography. The native enzyme was a single polypeptide chain with a molecular weight of about 65,000 and an isoelectric point at pH 4.8. The optimum pH for esterase activity was 9.0, and its pH stability range was 5.0–10.5. The optimum temperature for its activity was 60°C. The esterase had a half-life of 28 h at 50°C, 20 h at 60°C and 16 h at 65°C. It showed the highest activity on tributyrin, with little or no activity toward long-chain (12–20 carbon) fatty acid esters. The enzyme displayed K_m and K_cat values of 0.357 mM and 8365/min, respectively, for tributyrin hydrolysis at pH 9.0 and 60°C. Cyclodextrin (α, β, and γ), Ca²⁺, Co²⁺, Mg²⁺ and Mn²⁺ enhanced the esterase activity, and Zn²⁺ and Fe²⁺ acted as inhibitors of the enzyme activity. The enzyme activity was not affected by ethylenediaminetetraacetic acid, p-chloromercuribenzoate andN-bromosuccinimide. This paper was presented in part at the 82nd Annual Meeting and Exposition of the American Oil Chemists’ Society, held May 12–15, 1991, in Chicago, Illinois.     

Characterization and Immobilization of Marine Algal 11-Lipoxygenase from Ulva fasciata

The lipoxygenase (LOX) of the marine green alga Ulva fasciata was purified and immobilized in order to improve the stability and reusability. The algal LOX was partially purified by fractionation with 35–55% saturation of ammonium sulfate and MacroPrep high Q anion exchange chromatography. The LOX was purified ten times using linoleic acid (C_18:2) or arachidonic acid (C_20:4) as substrate, the Michaelis constant (K _m) of LOX was 117.6, 31.3 μM, and maximum velocity (V _max) was 12.8, 23.3 μmol hydroperoxy fatty acid/min-mg protein, respectively. The algal LOX showed the highest activity towards C_18:4 followed by C_20:4, C_18:2 and methyl ester of C_18:4. LOX activity increased up to 10.5 times with increased concentration of Triton X-100 in the extraction medium reaching an optimum at 0.05%. Calcium chloride, glutathione and phenylmethylsulphonyl fluoride were found effective protectants to LOX during purification. Hydroperoxyeicosatetraenoic acid (HpETE) formed from arachidonic acid catalyzed by this purified algal LOX was reduced and identified as 11-hydroxy-5,8,12,14-eicosatetraenoic acid (11-HETE) by NP-HPLC and GC–MS. This algal 11-LOX was immobilized in alginate beads. The stability was sevenfold greater than that of the unbound lipoxygenase at 4 °C in 0.05 M Tris–HCl buffer (pH 7.5). This is the first report on immobilization of a marine algal lipoxygenase with a view to its potential role in seafood flavor formation.     

Synthesis and surface properties of chemodegradable anionic surfactants: Diastereomeric (2-n-alkyl-1,3-dioxan-5-yl) sulfates with monovalent counter-ions

Sodium, potassium and ammonium cis- and trans-(2-n-alkyl-1,3-dioxan-5-yl) sulfates 6–8 (alkyl: n-C₉H₁₉, 6a–8a, and n-C₁₁H₂₃, 6b–8b) were synthesized in a reaction of aliphatic aldehydes 1a,b with glycerol 2 followed by separation in high yields of individual geometric isomers of cis-and trans-2-n-alkyl-5-hydroxy-1,3-dioxanes, cis-3a,b and trans-3a,b, followed by sulfation with sulfur trioxide-pyridine complex, and finally neutralization with NaOH, KOH, and NH₄OH, respectively. Physical data of the compounds and some surface properties of 2-n-nonyl derivatives, such as critical micelle concentration (CMC), effectiveness of aqueous surface tension reduction (Π_CMC), surface excess concentration Γ_CMC, and the surface area demand per molecule (A_CMC), were determined. It was shown that the surface activity of these compounds is influenced both by their geometric structure and by the monovalent counter-ion.     

Environmentally acceptable drag-reducing surfactants for district heating and cooling

The aim of this work was to find drag-reducing surfactants for both district heating and cooling that are environmentally more acceptable than the organic salts of quaternary ammonium compounds that so far have dominated this application. Vortex inhibition of test solutions in glass beakers has been used to screen a large number of surfactant mixtures, various electrolytes, and temperatures. For the most interesting products, the pressure drop in a test loop was measured at various flow rates and temperatures.N-cetyl sarcosinate andN-stearyl betaine, the latter together with Na-dodecylbenzene sulfonate, give good drag reduction (DR) properties at 65–100 and 45–85°C, respectively. The sarcosinate is sensitive to pH changes, whereas the betaine-sulfonate complex is more robust. Ethoxylates of oleic acid monoethanolamide show good DR properties at 0–25°C. At an increased salt concentration, a mixture of this surfactant with an ethoxylated oleyl alcohol worked well. At a still higher salt concentration, e.g., sea water, a combination ofN-cetyl betaine and alkylbenzene sulfonate showed DR.     

Micellar and Surface Properties of Some Monomeric Surfactants and a Gemini Cationic Surfactant

The physicochemical and interfacial properties of the monomeric surfactants cetyltrimethyl ammonium bromide (CTAB), cetyltriphenyl phosphonium bromide (CTPB), tetradecyl triphenyl phosphonium bromide (TTPB), cetyldiethylethanol ammonium bromide (CDEEAB), cetyltrimethyl ammonium chloride (CTACl), tetradecyltrimethyl ammonium bromide (TTAB), and a gemini surfactant (C₁₆-3-C_16, 2Br⁻) at different pH (3.1, 7.0, and 7.75) have been investigated by conductivity and surface tension measurements at 300 K. The critical micellar concentration (CMC), degree of micellar ionization (α), surface excess concentration (Г_max), minimum surface area per molecule of surfactant (A _min), Gibbs free energy of micellization (∆G _m⁰), surface pressure at the CMC (π _CMC), and the Gibbs energy of adsorption (∆G _ads⁰) of the monomeric surfactants have also been determined. The CMC, α and Г_max, increase with increasing pH whereas A _min decreases.     

Vernonia galamensis: A rich source of epoxy acid

A percolation extraction ofVernonia galamensis seed, affording 38.6% of crude vernonia oil is described. The dark colored crude oil was degummed with water, treated with activated charcoal and bleached with a neutral agent, to give a light colored oil (Lovibond: 0.9 red, 3.5 yellow). Gas chromatographic/mass spectrometric analysis of the refined oil indicates a relative fatty acid composition of 79–81% vernolic (cis-12,13-epoxy-cis-9-octadecenoic) acid, 11–12% linoleic acid, 4–6% oleic acid, 2–3% stearic acid, 2–4% palmitic acid, and a trace amount of arachidic acid.     

Biosurfactant production by <Emphasis Type="Italic">Bacillus coagulans</Emphasis>

A new biosurfactant producer, Bacillus coagulans, was isolated from soil. Its 24-h-old culture broth had a low surface tension (27–29 mN/m). Optimization of cell growth of this bacterium led to maximal biosurfactant production with glucose or starch as the organic carbon source, a pH in the range 4.0–7.5, and incubation temperatures from 20 to 45°C. The crude biosurfactants obtained after neutralization and lyophilization of the acid precipitate yielded a minimal aqueous solution surface tension value of 29 mN/m and an interfacial tension value of 4.5 mN/m against hexadecane. The critical micelle concentration of the crude biosurfactants was 17 mg/L. Addition of NaCl to the aqueous solution of the crude product caused lowering of surface tension at both the aqueous solution-air and aqueous solution-n-hexadecane interfaces. These results indicate that the biosurfactants obtained have potential environmental and industrial applications and may have uses in microbially enhanced oil recovery.     

Surface and thermodynamic parameters of mixed N-dodecyl N-trimethylsilane ammonium chloride with isooctyl phenol ethoxylate

Isooctyl phenol ethoxylate was mixed with N-dodecyl-N-trimethylsilane ammonium chloride and N-hexadecyl-N-trimethylsilane ammonium chloride, respectively. The surface tensions of aqueous solutions of cationic-nonionic mixtures were measured as a function of nonionic concentration at 35°C with constant cationic concentration. By using the Hutchinson method, the surface excess concentrations, surface mole fractions, and interaction parameters of binary surfactant mixtures were calculated. A synergistic effect for reducing surface tension was observed for adsorption at the surface at low concentrations of N-dodecyl-N-trimethylsilane ammonium chloride.     

Growth and eicosapentaenoic acid production byPhaeodactylum tricornutum in batch and continuous culture systems

Maximum specific growth rate (μ_max) ofPhaeodactylum tricornutum increased with increasing culture reactor surface-to-volume ratio. Values for μ_max of 0.647, 0.377 and 0.339 day⁻¹ were observed for the 75-mL tube, 5.6-L tank and the 16-L tank, respectively. Higher biomass was achieved in the 75-mL batch culture tube under continuous light as compared with light cycle conditions. Palmitic acid, palmitoleic acid and eicosapentaenoic acid (EPA) accounted for over 60% of total fatty acids in the batch tube culture, with EPA content increasing to a maximum after three days. In chemostat cultures, run at dilution rates of 0.15 day⁻¹ (0.45 of μ_max) and 0.3 day⁻¹ (0.9 of μ_max), cell concentration reached a steady state of 2.18 and 0.7 g/L, respectively, while contents of EPA per liter of culture at steady state were 100.9 and 82.5 mg/L, respectively. At both dilution rates, EPA content of total fatty acids was the same (35.0–35.2%). At a dilution rate of 0.3 day⁻¹, the continuous culture system manifested productivities of 0.51 g/L/d and 25.1 mg/L/d for biomass and EPA, respectively.     

Screening study of spray solution parameters for depositing cerium-based conversion coatings on Al alloy 2024-T3

Cerium-based conversion coatings were deposited on aluminum alloy 2024-T3 by a spray process using a solution containing cerium chloride, hydrogen peroxide, and gelatin. As deposited coatings were composed of hydrated cerium oxide and were post-treated in a phosphate solution to improve corrosion performance. Coating solution parameters, including the pH (1–2.5), cerium chloride concentration (0.025–0.125 M), and hydrogen peroxide content (0–1.2 M), were varied to investigate the effect(s) of solution parameters on the corrosion performance of the post-treated coatings. Results indicated that thickness of coatings deposited from solutions with different pH values were similar, while coating thickness increased with increasing concentration of cerium chloride and hydrogen peroxide in the solutions. Electrochemical impedance spectroscopy and observations of the surface appearances of the coatings indicated that coatings deposited from solutions with a pH 2, a cerium concentration of 0.1 M, and a hydrogen peroxide concentration of 0.8 M exhibited the best corrosion resistance.     

Synthesis and Properties of Organosilicon Quaternary Salts Surfactants

Diethyl-benzyl-[3-methyldimethoxyl)]silpropyl ammonium chloride (DEBSAC) was synthesized from N,N-diethyl-aminopropyl-methyldimethoxysilane (DEAPMDES) and benzyl chloride. Diethyl-2,3-epoxypropyl-[3-methyldimethoxyl)] silpropyl ammonium chloride (DEEPSAC) was synthesized from DEAPMDES and epoxy chloropropane. Trimethyl-[3-methyldimethoxyl)] silpropyl ammonium chloride (TMSAC) was synthesized from trimethylamine and γ-chloropropylmethyldimethoxysilane (CPMDMS). The products were characterized by ¹H NMR, ¹³C NMR, and IR. The surface tension measurements showed that the organosilicon quaternary salts exhibit a lower surface tension and a lower critical micelle concentration (CMC) than the hydrocarbon counterparts. The plate count method experiments illustrated that DEEPSAC has a better degree of antibacterial activity against Escherichia coli than DEBSAC. The solubilizing effects of the organosilicon quaternary salts on organosilicone were studied by pseudo-ternary phase diagrams of synthesized products/cosurfactant(n-butanol)/water/octamethylcyclotetrasiloxane. The solubilizing activities decreased in the order of TMSAC > DEBSAC > DEEPSAC.     

The effect of two isomeric octadecenoic acids on alkyl diacyl glycerides and neutral glycosphingolipids of Novikoff hepatoma cells

The addition of 150 μg ofcis-6-octadecenoic acid (6–18∶1) per milliliter to the growth medium of Novikoff hepatoma cells reduced the rate of multiplication of these cells, whereas the same level ofcis-9-octadecenoic acid (9–18∶1) was without effect. The quantity of alkyl diacyl glycerols in cells grown in medium containing 100 μg/ml of either 6–18∶1 or 9–18∶1 was reduced to about 10% of that observed in cells grown in unsupplemented media. The presence of 6–18∶1 acid led to an increase in the concentration of the dihexosyl ceramide and a corresponding decrease in that of the monohexosyl ceramides of the cells; whereas the presence of 9–18∶1 resulted in an increase in the relative amount of tetraglycosyl ceramide and corresponding reductions in those of mono- and dihexosyl ceramide. Possible causes for these changes are discussed.