Lipid Production by Arctic Microalga Chlamydomonas sp. KNF0008 at Low Temperatures

A lipid-producing microalga, Chlamydomonas sp. KNF0008, collected from the Arctic was capable of growing at temperatures ranging from 4 to 20 °C, and the highest cell density was measured at 15 °C and 100 μmol photons m^?2 s^?1 light intensity under continuous shaking and external aeration. KNF0008 showed the elevated accumulation of lipid bodies under nitrogen-deficient conditions, rather than under nitrogen-sufficient conditions. Fatty acid production of KNF0008 was 4.2-fold (104 mg L^?1) higher than that of C. reinhardtii CC-125 at 15 °C in Bold’s Basal Medium. The dominant fatty acids were C16:0, C16:4, C18:1, and C18:3, and unsaturated fatty acids (65.69%) were higher than saturated fatty acids (13.65%) at 15 °C. These results suggested that Arctic Chlamydomonas sp. KNF0008 could possibly be utilized for production of biodiesel during periods of cold weather because of its psychrophilic characteristics.

     

A Novel Labeling Reagent of 2-(12-Benzo[b]acridin-5-(12H)-yl)-acetohydrazide for Determination of Saturated and Unsaturated Fatty Acids in Traditional Chinese Herbs by HPLC-APCI-MS

A new fluorescence labeling reagent 2-(12-benzo[b]acridin-5(12H)-yl)-acetohydrazide (BAAH) has been designed for fatty acids labeling. Eleven fatty acids containing seven saturated and four unsaturated fatty acids were used to evaluate the analytical potential of this reagent. The labeling reaction of BAAH with fatty acids was completed at 85 °C for 60 min using 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC·HCl) as the condensing agent. Separation of the derivatized fatty acids was carried out on a reversed-phase Thermo Hypersil Gold C18 column (4.6 mm × 250 mm, 5 μm) in combination with a gradient elution with a good baseline resolution. The fluorescence excitation and emission wavelengths were set at λ_ex 280 and λ_em 510 nm, respectively. The identification was carried out by the online APCI-MS in positive-ion detection mode. Linear correlation coefficients for all fatty acid derivatives were of >0.9994. Detection limits, at a signal-to-noise ratio of 3:1, were 3.89–12.5 nmol L⁻¹ for the labeled fatty acids. The developed method was successfully applied to the accurate determination of fatty acids in five traditional Chinese herbs with satisfactory results.

     

Thermal stability of the silica-aminopropylsilane-polyimide interface

Thermal degradation of the silica–aminopropylsilane–amic acid/imide interface was studied by modifying a high-surface-area, neutral silica gel with a number of substituted aminopropylsilanes (APS). These substrates were reacted further with phthalic anhydride or aromatic amic acid monomers and the thermal decomposition of the adsorbed/reacted materials was monitored by thermogravimetric analysis (TGA) and infrared (IR) spectroscopy. The 3-aminopropyltriethoxysilane/poly[N,N′-(p,p′-oxydiphenylene)pyromellitimide] interface was also evaluated by this method. Comparison clearly distinguishes the thermal decomposition of surface-bound APS from surface-bound alkylphthalimides, the adhesion product of alkylamines and aromatic amic acids. Alkylamine imidization with the elimination of aromatic amine (analogous to polymer scission) and the decomposition of the surface-bound imide are shown in the amic acid TGA profiles. This imidization and the accompanying aniline elimination begin at about 130°C, under nitrogen, to form the surface alkyl imide which slowly decomposes at 400°C. TGA analysis indicates that the surface-bound imide undergoes minimal degradation under nitrogen at 370 ± 10°C; temperatures above this threshold range produce changes in the APS–imide interface.     

Development of a Temperature Shift Strategy for Efficient Docosahexaenoic Acid Production by a Marine Fungoid Protist, <Emphasis Type="Italic">Schizochytrium</Emphasis> sp. HX-308

Temperature was one of the important environmental factors affecting the biosynthesis of docosahexaenoic acid (DHA; C22:6, ω−3). Generally, a low temperature would slow the strain growth, but promote the accumulation of unsaturated fatty acids. According to this information, the effects of temperature and different two-stage temperature shifting strategies on fatty acid production and DHA content of the marine fungoid protist, Schizochytrium sp. HX-308, were investigated in this study. Finally, the highest DHA percentage was up to 51.98% (per total fatty acids) with the DHA production of 6.05% (per dry cell weight), which was obtained with the method of shifting temperature from 30 °C for 32 h to 20 °C for 12 h.     

The Role of Lignocellulosic Composition and Residual Lipids in Empty Fruit Bunches on the Production of Humic Acids in Submerged Fermentations

The aim of this research was to study the production of humic acids (HA) by Trichoderma reesei from empty fruit bunches (EFBs) of palm oil processing, with a focus on the effects of lignocellulosic content and residual lipids. EFBs from two different soils and palm oil producers were previously characterized about their lignocellulosic composition. Submerged fermentations were inoculated with T. reesei spores and set up with or without residual lipids. The results showed that the soil and the processing for removal of the palm fresh fruits were crucial to EFB quality. Thus, EFBs were classified as type 1 (higher lignocellulosic and fatty acids composition similar to the palm oil and palm kernel oil) and type 2 (lower lignocellulosic content and fatty acids composition similar to palm oil). Despite the different profiles, the fungal growth was similar for both EFB types. HA production was associated with fungal growth, and it was higher without lipids for both EFBs. The highest HA productivity was obtained from type 1 EFB (approximately 90 mg L⁻¹ at 48 h). Therefore, the lignocellulosic composition and the nature of the residual lipids in EFBs play an important role in HA production by submerged fermentation.

     

Radio-gas-chromatographische Kontrolle des 14C-Acetateinbaus in die Phosphatidfettsäuren der Sojabohnen

Germinating soybeans (glycine max.L.) can be employed to give a good yield of labelled phosphatides. The incorporation of ¹⁴C-acetate into fatty acids of the phosphatides, which under optimal conditions (35°C, 3000 Lux) is about 15%, was radio-gas-chromatographically controlled. It was found that exposure to light causes an increase in the formation of unsaturated fatty acids. Under the influence of a pure oxygen atmosphere at 16°C the phosphatides obtained have about 50% of the ¹⁴C-radioactivity in the oleic acid and 20% in linolic acid. This gives the preconditions necessary to obtain natural phosphatides with unequal distribution of radioactivity in the saturated and unsaturated fatty acids.

     

Determination of Fatty Acids (C1 – C10) from Bryophytes and Pteridophytes

A simple and mild method for the determination of fatty acids (C₁ – C₁₀) based on a condensation reaction using 7-aminonaphthalene-1,3-disulfonic acid (ANDSA) as labeling reagent with capillary zone electrophoresis has been developed. The detection was performed with a diode array detector at 254 nm. A 58.5 cm × 50 μm i.d. (50 cm effective length) untreated fused-silica capillary was used. To optimize the separation conditions, the background electrolyte concentration, column temperature, voltage and other factors were evaluated. The optimal separation conditions were as follows: 30 mmol L⁻¹ borate buffer (pH 9.5), 15 mmol L⁻¹ β-CD, temperature at 20 °C, pressure 50 mbar and injection time 8 s. Under the established conditions, 10 fatty acid derivatives could be well-separated within 17 min. The linearity was in the range of 0.07–5.0 μmol L⁻¹. Detection limits (at a signal-to-noise ratio of 3) were in the range of 0.027–0.042 μmol L⁻¹. The fatty acids from the extracted Funaria Hedw. and Selaginella samples were determined with satisfactory results.

     

Microbial Lipid Production from Enzymatic Hydrolysate of Pecan Nutshell Pretreated by Combined Pretreatment

Biodiesel is a fuel composed of monoalkyl esters of long-chain fatty acids derived from renewable biomass sources. In this study, biomass waste pecan nutshell (PS) was attempted to be converted into microbial oil. For effective utilization of PS, sequential pretreatment with ethylene glycol–H₂SO₄–water (78:2:20, wt:wt:wt) at 130 °C for 30 min and aqueous ammonia (25 wt%) at 50 °C for 24 h was used to enhance its enzymatic saccharification. Significant linear correlation was obtained about delignification-saccharification (R ² = 0.9507). SEM and FTIR results indicated that combination pretreatment could effectively remove lignin and xylan in PS for promoting its enzymatic saccharification. After 72 h, the reducing sugars from the hydrolysis of 50 g/L pretreated PS by combination pretreatment could be obtained at 73.6% yield. Using the recovered PS hydrolysates containing 20 g/L glucose as carbon source, microbial lipids produced from the PS hydrolysates by Rhodococcus opacus ACCC41043. Four fatty acids including palmitic acid (C16:0; 23.1%), palmitoleic acid (C16:1; 22.4%), stearic acid (C18:0; 15.3%), and oleic acid (C18:1; 23.9%) were distributed in total fatty acids. In conclusion, this strategy has potential application in the future.     

Isolation,Identification and High-Throughput Screening of Neutral Lipid Producing Indigenous Microalgae from South African Aquatic Habitats

Exploring indigenous microalgae capable of producing significant amounts of neutral lipids through high-throughput screening is crucial for sustainable biodiesel production. In this study, 31 indigenous microalgal strains were isolated from diverse aquatic habitats in KwaZulu-Natal, South Africa. Eight superior lipid-producing strains were selected for further analysis, based on Nile red fluorescence microscopy screening. The microalgal isolates were identified to belong to the genera Chlorella, Neochloris and Chlamydomonas via morpho-taxonomic and molecular approach by 18S rRNA gene sequencing. Chlorella vulgaris PH2 had the highest specific growth rate (μ) and lowest doubling time of 0.24 day⁻¹ and 2.89 ± 0.05 day⁻¹, respectively. Chlorella vulgaris T4 had the highest biomass productivity of 35.71 ± 0.03 mg L⁻¹day⁻¹. Chlorella vulgaris PH2 had the highest lipid content of 34.28 ± 0.47 and 38 ± 9.2% (dcw) as determined by gravimetric analysis and the sulfo-phospho-vanillin (SPV) method, respectively. Chlorella vulgaris PH2 exhibited a high content of saturated fatty acids, while Chlorella sp. T4 exhibited a high total content of saturated and monounsaturated fatty acids with a low content of polyunsaturated fatty acids. The preponderance of neutral lipids suggests that Chlorella sp. T4 is a suitable candidate for biomass feedstock for biodiesel production.

     

Analysis of Free Fatty Acids on the Fingertips by High Performance Liquid Chromatography

Abstract

This investigation studied the efficiency of high performance liquid chromatography in the determination of free fatty acids present on the fingertips, and assessed the quantitative relationship between skin fatty acids and the degree of microbial contamination. Automated surgical scrub was utilized to eliminate the microbial contamination.

The high performance liquid chromatography provided excellent separation of skin fatty acids for evaluation with the bacterial counts. The fatty acid peaks identified ranged in chain length from C12 through C32. All the fatty acids evaluated showed positive correlation with the bacterial counts with the exception of one acid which had an inverse relationship but none were statistically significant. Finally, the surgical scrub chromatograms showed that the straight chain acids C19 and C21 were lower in concentration than C23 and C25; also, C26 was lower in concentration than C28 and C30.

It was evident from the data that fatty acids which have been shown to be bacteriostatic in vitro do not demonstrate the same property on the fingertips. The finding of lower concentration of C19, C21, and C26 than longer chain acids is inconsistent with simple two carbon addition, and indicates there is possible branching at these points in metabolic pathway of fatty acid synthesis.     

Determination of cuticular and internal fatty acids of Chorthippus brunneus males and females using HPLC‐LLSD and GC–MS

Insects are of growing significance in veterinary medicine and human healthcare; therefore, an understanding of their biology is very important. The cuticular and internal fatty acid compositions of Chorthippus brunneus males and females have been studied for the first time. The lipids of males and females were separated into classes of compounds using high‐performance liquid chromatography with a laser light scattering detector. The free fatty acid (FFA) fractions obtained by HPLC were silylated and then analyzed by GC–MS. The cuticular lipids of males contained 15 saturated, four unsaturated with even‐numbered and two unsaturated with odd‐numbered carbon chains, FFAs ranging from C8 to C25. The major free fatty acids in males were C16 (20.8%), C18:2 (8.5%), C18:1 (32.9%) and C18 (24.4%). The cuticular lipids of females contained 17 saturated, four monounsaturated and two diunsaturated free fatty acids ranging from C8 to C30. The major cuticular fatty acids in females were C16 (25.1%), C18:2 (6.2%), C18:1 (23.7%) and C18:0 (33.2%). The internal FFAs of males consisted of 20 compounds ranging from C8 to C26. Four of these compounds were detected as major compounds: C16 (14.1%), C18:2 (21.6%), C18:1 (38.0%) and C18 (22.5%). Among 18 internal free fatty acids of females, C16 (22.3%), C18:2 (10.9%), C18:1 (40.2%) and C18 (20.5%) were the most abundant compounds. The following cuticular fatty acids present in the lipids of females were absent in the lipids of males: C26, C27 and C30. On the other hand, only C24 was absent from the cuticular lipids of females. Only C10 and C24 internal fatty acids present in the lipids of males were absent in the lipids of females. Copyright © 2016 John Wiley & Sons, Ltd.     

Gibberellin Promotes Cell Growth and Induces Changes in Fatty Acid Biosynthesis and Upregulates Fatty Acid Biosynthetic Genes in Chlorella vulgaris UMT-M1

Microalgae lipids and oils are potential candidates for renewable biofuels and nutritional inventions. Recent studies from our lab have shown that two plant hormones, auxin and jasmonic acid, influence microalgae growth and fatty acid accumulation. Therefore, in this study, a high oil-producing strain Chlorella vulgaris UMT-M1 was selected for hormonal study using gibberellin (GA). Exogenous GA₃ was applied to early stationary culture of C. vulgaris UMT-M1. Results showed that GA₃ gradually increases the cell density of C. vulgaris to up to 42% on days after treatment (DAT)-8 and also capable of delaying the algal senescence. However, the increment in cell density did not enhance the total oil production albeit transient modification of fatty acid compositions was observed for saturated (SFA) and polyunsaturated (PUFA) fatty acids. This illustrates that GA₃ only promotes cell division and growth but not the oil accumulation. In addition, application of GA₃ in culture medium was shown to promote transient increment of palmitic (C16:0) and stearic (C18:0) acids from DAT-4 to DAT-6 and these changes are correlated with the expression of β-ketoacyl ACP synthase I (KAS I) gene.

     

A mutant of Escherichia coli K12 exhibiting varying ultraviolet sensitivities depending on the temperature of incubation after irradiation

Abstract— A mutant, URT-43, was isolated from E. coli C600 dar⁺. The mutant has a characteristic feature in that its sensitivity to ultraviolet (u.v.) light is greatly influenced by the temperature at which irradiated bacteria are incubated. On the basis of dose-reduction factor, URT-43 is approximately ten times more sensitive at 42° than at 30°C, even though unirradiated bacteria are not thenno-sensitive, The mutant could not repair u.v.-irradiated bacteriophage Λ_vir in the dark either at 30° or at 42°C, indicating that it is defective in host-cell reactivation. In contrast, the same bacteriophage was reactivated in preirradiated URT-43 if the host-bacteriophage complex was plated at 30° but there was no reactivation at 42°C. Therefore u.v.reactivation was positive at 30° but negative at 42°C. The induction of prophage by URT-43(Λ_h) was achieved by much lower doses of U.V. light than that required for the induction of lysogenic wild type bacteria. Experiments were performed in which irradiated URT-43 was first incubated for various periods in liquid media and plated both at 30° and 42°C. It was found that irradiated bacteria came to be resistant to subsequent plating at 42° only when they were preincubated in the liquid medium containing necessary amino acids and at 30°C. Since this phenomenon was completely inhibited by chloramphenicol, the process seemed to require de novo protein synthesis. An hypothesis was proposed that there are at least two independent dark-repair mechanisms in E. coli; one is responsible for host-cell reactivation and the other is responsible for U.V. reactivation.     

HPLC Analysis of Nonionic Sufactants. V. Ethoxylated Fatty Acids

Abstract

High performance liquid chromatography technique was used in order to achieve separation and identification of product composition of nonionic surfactants of ethoxylated fatty acids.

Lichrosorb SI-60 (10μm) column, under gradient elution of mixture of isopropanol, methanol and n-hexane (50°C) and UV detector at 220 nm, were used for best separation of ethylene oxide (EO) adducts of fatty acids consisting of up to 20 EO units.

No derivatization of the compounds was needed. An improved baseline, in spite of gradient elution, was achieved by adding negligible amounts of anthracene to the eluents.

Brominated ethoxylated fatty acids resulting from addition of bromine to the double bond of the hydrophobic chain were also separated without a need for change in elution conditions or derivatization.     

Impact of High Temperature on Ethanol Fermentation by Kluyveromyces marxianus Immobilized on Banana Leaf Sheath Pieces

Ethanol fermentation was carried out with Kluyveromyces marxianus cells at various temperatures (30, 35, 40, and 45 °C). Fermentation performance of the immobilized yeast on banana leaf sheath pieces and the free yeast were evaluated and compared. Generally, ethanol production of the immobilized and free yeast was stable in a temperature range of 30–40 °C. Temperature of 45 °C restricted yeast growth and lengthened the fermentation. The immobilized yeast demonstrated faster sugar assimilation and higher ethanol level in the fermentation broth in comparison with the free yeast at all fermentation temperatures. Change in fatty acid level in cellular membrane was determined to clarify the response of the free and immobilized yeast to thermal stress. The free cells of K. marxianus responded to temperature increase by increasing saturated fatty acid (C16:0 and C18:0) level and by decreasing unsaturated fatty acid (C18:1 and C18:2) level in cellular membrane. For fermentation at 40 °C with immobilized cells of K. marxianus, however, the changes were not observed in both saturated fatty acid (C16:0) and unsaturated fatty acid (C18:1 and C18:2) level.     

Temperature Dependence in the Synthesis of SBA-16-Type Mesoporous Silica with Pluronic F68 Block Copolymer

By adjusting the local effective surfactant packing parameter through synthesis temperature, highly ordered SBA-16-type mesoporous silica materials have been synthesized by templating with a nonionic triblock copolymer Pluronic F68 in strongly acidic conditions at temperature 30～40°C with the addition of K₂SO₄. The prepared SBA-16-type mesoporous silica materials having Im3m cubic mesostructure were proved by the well-defined x-ray diffraction patterns combined with transmission electron microscopy. Scanning electron microscopy indicated that a transformation from faced-sphere to faced-polyhedron shape morphologies could be induced with increasing of the synthesis temperature. The nitrogen adsorption–desorption analysis revealed that the mean pore size (5.50～6.13 nm) of the prepared materials increased with increasing synthesis temperature. However, when the synthesis temperature exceeded 46°C, only disordered mesoporous silca was obtained. Our synthesis strategies by adjusting the local effective surfactant packing parameter through synthesis condition, even in a narrow range, would be used not only to optimize the synthesis conditions of reported mesoporous silca, but also to fabricate new mesoporous silica materials with well-ordered channel and anticipated morphologies.     

Determination of 30 Free Fatty Acids in Two Famous Tibetan Medicines by HPLC with Fluorescence Detection and Mass Spectrometric Identification

A simple and sensitive high-performance liquid chromatographic (HPLC) method with fluorescence detection and mass spectrometric identification has been developed for analysis of 30 long-chain and short-chain free fatty acids (FFAs). The fatty acids were derivatized to their esters with 1-[2-(p-toluenesulfonate)ethyl]-2-phenylimidazole-[4,5-f]-9,10-phenanthrene (TSPP) in N,N-dimethylformamide (DMF) at 90 °C with anhydrous K₂CO₃ as catalyst. A mixture of C₁–C₃₀ fatty acids was completely separated within 60 min by gradient elution on a reversed-phase C₈ column. Qualitative identification of the acids was performed by atmospheric-pressure chemical ionization mass spectrometry (APCI–MS) in positive-ion mode. The fluorescence excitation and emission wavelengths were 260 and 380 nm, respectively. Quantitative determination of the 30 acids in two Tibetan medicines Gentiana straminea and G. dahurica was performed. The results indicated that the medicines contained many FFAs. Linear correlation coefficients for the FFA derivatives were >0.9991. Relative standard deviations (RSDs, n = 6) for the fatty acid derivatives were <3%. Detection limits (at a signal-to-noise ratio of 3:1) were 3.1–38 fmol. When the fatty acid derivatives were determined in the two real samples results were satisfactory and the sensitivity and reproducibility of the method were good.     

Combination of Silica Sol and Potassium Silicate via Isothermal Heat Conduction Microcalorimetry

Isothermal heat conduction microcalorimetry was utilized as a novel characterization method to investigate the polymerization processes of silica with both thermodynamic and kinetic parameters when the combination of silica sol and potassium silicate was stirred at temperatures of 25.0, 35.0, and 45.0°C. The silica polymerization was characterized by the greater enthalpy change at each higher temperature and by the reaction orders of the silica sol and potassium silicate, which varied rapidly, instantaneously, and constantly from low to high all the time, up and down in an alternate manner. When the reaction order of the silica sol and potassium silicate was 3.0, the maximum rate constant occurred at 25.0°C (k=1.22×10^?4mol^?2·dm⁶·s^?1). The two temperature regions (25.0–35.0°C region with a faster rate and 35.0–45.0°C region with a lower rate) reflected a two‐stage oligomerization of silica monomers with different oligomers formed in a two‐step anionic mechanism. The measurements of particle size and pH value showed that the colloidal particles in the mixed silica sol and potassium silicate first dissolved, then "active" silica in the potassium silicate redeposited to make a distinct particle size distribution (Z‐average size, 33.0–14.9 nm at 25.0°C) influenced both by pH value (9.82–11.97 at 25.0°C) and the mass fraction (53, 65, 75, and 85 mass/%) of the silica sol in the mixture. The processes of combination of the silica sol and potassium silicate did not result from acid‐base neutralization reactions but from a complex polymerization of the "active" silica components which relate to silica monomers oligomerization with heat evolved (the total enthalpy changes, 1.6234–3.3882 J).     

Crystallinity of Dynasan<Superscript>®</Superscript>114 and Dynasan<Superscript>®</Superscript>118 matrices for the production of stable Miglyol<Superscript>®</Superscript>-loaded nanoparticles

This study focuses on the physicochemical characterization of lipid materials useful for the production of the so-called solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC). The chosen lipids were Dynasan^®114 (glyceril trimyristate) and Dynasan^®118 (glyceril tristearate) as solid lipids (SL), melting temperature above 80 °C, and Miglyol^®812 (caprylic/capric triglyceride) and Miglyol^®840 (propylene glycol dicaprylate/dicaprate) as liquid lipids (LL), crystallizing below ?15 °C. Raw lipids (pure or SL:LL mixtures) were analyzed by differential scanning calorimetry (DSC), wide angle X-ray diffraction (WAXD), and Polarized Light Microscopy (PLM), before and after tempering at 80 °C for 1 h. The selected SL:LL combination was 70% (Dynasan^®114 and 118) and 30% (Miglyol^®812 and 840) for the production of SLN and NLC by high-pressure homogenization (HPH), respectively. Particles with a mean size of 200 nm (polydispersity index <0.329) and zeta potential of ?15 mV were obtained, and their long-term stability was confirmed for 3 months of storage at 7 °C.     

Formation of the main degradation compounds from arabinose,xylose, mannose and arabinitol during pyrolysis

The thermochemical behaviour of sugars (D- and DL-arabinose, D- and DL-xylose and D-mannose) and sugar alcohol (D- and DL-arabinitol) was investigated by TG and pyrolysis-gas chromatography with mass-selective detection (Py-GC/MSD). The temperature of pyrolysis was 500 and 550°C. The TG-curves were measured both in air and nitrogen atmospheres, from 25 to 700°C with the heating rate of 2°C min^-1. In each case, the main pyrolysis products were classified into the following compound groups: (i) furanes, (ii) pyranes, (iii) cyclopentanes, (iv) cyclohexanes, (v) anhydroglucopyranoses, (vi) dianhydroglucopyranoses and (vii) saturated fatty acids. For example, the main peaks of the chromatograms of pentoses (arabinose, xylose), hexose (mannose) and sugar alcohols (arabinitols) were different. The greatest peak of pentoses in gas-chromatogram was 2-furancarboxaldehyde and that of hexose was (2H)-furan-3-one. The greatest peak of arabinitols at pyrolysis temperature of 500°C was furan methanol and at 550°C a-angeligalactone. 5-hydroxymethyl-2-furan carboxaldehyde was found only in the pyrolysis of D-mannose (hexose). The former study showed that it was not found in pyrolysis of pentoses. The amount of CO₂ and H₂O was not determined. This revised version was published online in July 2006 with corrections to the Cover Date.