Wax Esters Research Articles

Influence of aphotic haloclines and euxinia on organic biomarkers and microbial communities in a thalassohaline and alkaline volcanic crater lake.

Studies on microbial communities, and their associated organic biomarkers, that are found thriving in the aphotic euxinic waters in modern stratified ecosystems are scarce compared to those undertaken in euxinic photic zones. The Dziani Dzaha (Mayotte, Indian Ocean) is a tropical, saline, alkaline crater lake that has recently been presented as a modern analog of Proterozoic Oceans due to its thalassohaline classification (having water of marine origin) and specific biogeochemical characteristics. Continuous intense photosynthetic production and microbial mineralization keep most of the water column permanently aphotic and anoxic preventing the development of a euxinic (sulfidic and anoxic) photic zone despite a high sulfide/sulfate ratio and the presence of permanent or seasonal haloclines. In this study, the molecular composition of the organic matter in Lake Dziani Dzaha was investigated and compared to the microbial diversity evaluated through 16S rRNA gene amplicon sequencing, over two contrasting seasons (rainy vs. dry) that influence water column stratification. Depth profiles of organic biomarker concentrations (chlorophyll-a and lipid biomarkers) and bacterial and archaeal OTU abundances appeared to be strongly dependent on the presence of aphotic haloclines and euxinia. OTU abundances revealed the importance of specific haloalkaliphilic bacterial and archaeal assemblages in phytoplanktonic biomass recycling and the biogeochemical functioning of the lake, suggesting new haloalkaline non-phototrophic anaerobic microbial precursors for some of the lipid biomarkers. Uncultured Firmicutes from the family Syntrophomonadaceae (Clostridiales), and Bacteroidetes from the ML635J-40 aquatic group, emerged as abundant chemotrophic bacterial members in the anoxic or euxinic waters and were probably responsible for the production of short-chain n-alkenes, wax esters, diplopterol, and tetrahymanol. Halocline-dependent euxinia also had a strong impact on the archaeal community which was dominated by Woesearchaeota in the sulfide-free waters. In the euxinic waters, methanogenic Euryarchaeota from the Methanomicrobia, Thermoplasmata, and WSA2classes dominated and were likely at the origin of common hydrocarbon biomarkers of methanogens (phytane, pentamethyl-eicosenes, and partially hydrogenated squalene).

117 Evidence of defective fattyacidome and aminoacidome in sebaceous and non sebaceous skin areas in atopic dermatitis

Atopic dermatitis (AD) is a composite disease characterized by derangement of the skin permeability barrier (SPB). Little is known on the role played by the sebaceous gland (SG) activity in the SPB integrity in AD. Our objective was to investigate lipidomics profiles of sebaceous-type and epidermal-type free fatty acids (FFAs), together with levels of squalene, cholesterol, triglycerides (TGs), and wax esters (WEs) in sebum and stratum corneum (SC) specimens obtained from seborrheic and non-seborrheic areas in healthy and AD adults.

Fatty Acids Derivatives From Eukaryotic Microalgae, Pathways and Potential Applications.

The exploitation of petrochemical hydrocarbons is compromising ecosystem and human health and biotechnological research is increasingly focusing on sustainable materials from plants and, to a lesser extent, microalgae. Fatty acid derivatives include, among others, oxylipins, hydroxy fatty acids, diols, alkenones, and wax esters. They can occur as storage lipids or cell wall components and possess, in some cases, striking cosmeceutical, pharmaceutical, and nutraceutical properties. In addition, long chain (>20) fatty acid derivatives mostly contain highly reduced methylenic carbons and exhibit a combustion enthalpy higher than that of C14–20 fatty acids, being potentially suitable as biofuel candidates. Finally, being the building blocks of cell wall components, some fatty acid derivatives might also be used as starters for the industrial synthesis of different polymers. Within this context, microalgae can be a promising source of fatty acid derivatives and, in contrast with terrestrial plants, do not require arable land neither clean water for their growth. Microalgal mass culturing for the extraction and the exploitation of fatty acid derivatives, along with products that are relevant in nutraceutics (e.g., polyunsaturated fatty acids), might contribute in increasing the viability of microalgal biotechnologies. This review explores fatty acids derivatives from microalgae with applications in the field of renewable energies, biomaterials and pharmaceuticals. Nannochloropsis spp. (Eustigmatophyceae, Heterokontophyta) are particularly interesting for biotechnological applications since they grow at faster rates than many other species and possess hydroxy fatty acids and aliphatic cell wall polymers.

Influence of wax type on characteristics of oleogels from camellia oil and medium chain triglycerides

SummaryCamellia oil (CO) and medium chain triglycerides (MCTs), characterised by different carbon chain lengths, were gelled using beeswax (BW), candelilla wax (CLW) and carnauba wax (CRW). Critical concentrations, oil‐binding capacity (OBC), textural parameters, thermal properties, microscopic properties and infrared spectra were used to evaluate their differences. The carbon chain length of oil sources showed different results for the critical concentration of the three wax‐based gel oils at 25 °C, and the oleogel of MCT and CLW exhibited a minimum value (1%). The oleogels formed from CO and waxes with wax esters as the main components (BW and CRW) exhibit significantly lower OBC and textural value than the oleogels made from MCT with BW and CRW. Meanwhile, this is corroborated with the thermal behaviour and crystal morphology, and distribution of the oleogels. However, no differences in crystal types and generation of new functional groups were observed by X‐ray diffraction and Fourier‐transform infrared spectrometer, suggesting that the differences in the properties of wax‐based oleogels with different carbon chain lengths are attributed to the interactions between the waxes and the oil sources (physical interaction).

Effect of Biodegradable Hydrophilic and Hydrophobic Emulsifiers on the Oleogels Containing Sunflower Wax and Sunflower Oil.

The use of an appropriate oleogelator in the structuring of vegetable oil is a crucial point of consideration. Sunflower wax (SFW) is used as an oleogelator and displays an excellent potential to bind vegetable oils. The current study aimed to look for the effects of hydrophobic (SPAN-80) and hydrophilic (TWEEN-80) emulsifiers on the oleogels prepared using SFW and sunflower oil (SO). The biodegradability and all formulations showed globular crystals on their surface that varied in size and number. Wax ester, being the most abundant component of SFW, was found to produce fibrous and needle-like entanglements capable of binding more than 99% of SO. The formulations containing 3 mg of liquid emulsifiers in 20 g of oleogels showed better mechanical properties such as spreadability and lower firmness than the other tested concentrations. Although the FTIR spectra of all the formulations were similar, which indicated not much variation in the molecular interactions, XRD diffractograms confirmed the presence of β′ form of fat crystals. Further, the mentioned formulations also showed larger average crystallite sizes, which was supported by slow gelation kinetics. A characteristic melting point (Tm~60 °C) of triglyceride was visualized through DSC thermograms. However, a higher melting point in the case of few formulations suggests the possibility of even a stable β polymorph. The formed oleogels indicated the significant contribution of diffusion for curcumin release. Altogether, the use of SFW and SO oleogels with modified properties using biodegradable emulsifiers can be beneficial in replacing saturated fats and fat-derived products.

Prostaglandin E2 and F2α Alter Expression of Select Cholesteryl Esters and Triacylglycerols Produced by Human Meibomian Gland Epithelial Cells.

PGF2α analogs are commonly used to treat glaucoma and are associated with higher rates of meibomian gland dysfunction (MGD). The purpose of this study was to evaluate the physiological effects of PGF2α and PGE2 on immortalized human meibomian gland epithelial cells (HMGECs). HMGECs were immunostained for the 4 PGE2 receptors (EP1, EP2, EP3, and EP4) and 1 PGF2α receptor (FP) and imaged. Rosiglitazone-differentiated HMGECs were exposed to PGF2α and PGE2 (10-9 to 10-6 M) for 3 hours. Cell viability was assessed by an adenosine triphosphate-based luminescent assay, and lipid extracts were analyzed for cholesteryl esters (CEs), wax esters (WEs), and triacylglycerols (TAGs) by ESI-MSMSALL in positive ion mode by a Triple TOF 5600 Mass Spectrometer using SCIEX LipidView 1.3. HMGECs expressed 3 PGE2 receptors (EP1, EP2, and EP4) and the 1 PGF2α receptor (FP). Neither PGE2 nor PGF2α showed signs of cytotoxicity at any of the concentrations tested. WEs were not detected from any of the samples, but both CEs and TAGs exhibited a diverse and dynamic profile. PGE2 suppressed select CEs (CE 22:1, CE 26:0, CE 28:1, and CE 30:1). PGF2α dose dependently increased several CEs (CE 20:2, CE 20:1, CE 22:1, and CE 24:0) yet decreased others. Both prostaglandins led to nonspecific TAG remodeling. PGE2 and PGF2α showed minimal effect on HMGEC viability. PGF2α influences lipid expression greater than PGE2 and may do so by interfering with meibocyte differentiation. This work may provide insight into the mechanism of MGD development in patients with glaucoma treated with PGF2α analogs.

Tear Film Lipid Layer Structure: Self-Assembly of O-Acyl-ω-hydroxy Fatty Acids and Wax Esters into Evaporation-Resistant Monolayers.

In healthy eyes, the tear film lipid layer (TFLL) is considered to act as an evaporation resistant barrier, which prevents eyes from drying. Seeking to understand the mechanisms behind the evaporation resistance of the TFLL, we studied mixtures of lipid layer wax esters and O-acyl-ω-hydroxy fatty acids. Analyzing their self-assembly and biophysical properties led to new discoveries concerning the structure and function of the TFLL. We discovered how these lipids self-assemble at the air-water interface and form an efficient antievaporative barrier, demonstrating for the first time how the interaction of different tear film lipid species can improve the evaporation resistance compared with individual lipid classes on their own. These results provide a potential mechanism for the evaporation resistance of the lipid layer. In addition, the results serve as a base for the future development of improved dry eye treatments and other applications where the evaporation of water represents a significant challenge.

Multistage fractionation of pine bark by liquid and supercritical carbon dioxide

Multistage fractionation of pine bark was performed using subcritical and supercritical CO2 at increasing pressures and temperatures. In total, seven fractions were collected, which demonstrated different enrichments of families of compounds. In particular, subcritical CO2 yielded 41% of the total extract in which unsaturated fatty acids represented the most abundant family. The subsequent five supercritical steps increased the recovery of sterol esters, wax esters and resin acids at higher temperatures and pressures, reaching 80% of the total extractable mass. In the last step, using ethanol as a co-solvent, an additional 20% of extract was recovered, which was enriched with phenolics and glycerol. A full characterisation of the extracts was accomplished by high-temperature GC–MS/FID using four internal standards, which were representative of the main classes of compounds contained in the pine bark extract.

Lipidomics of facial sebum in the comparison between acne and non-acne adolescents with dark skin

Lipidomics is advantageous in the study of sebum perturbations occurring in acne. An extended evaluation of the sebum lipid profiles in acne-prone sebaceous areas is lacking in dark skin. Yet, there is a void space in understanding how the building blocks of sebum lipids, i.e. individual fatty acids (FAs), are intertwined with acne-prone skin. We aimed to determine the sebum lipidome in facial areas of adolescents with and without acne in Nigeria. A cross-sectional analytical study was conducted in 60 adolescents/young adults divided in 30 acne patients (15F, 15M) and 30 age and sex-matched controls. Sebum samples obtained from foreheads and cheeks were analysed separately by gas chromatography–mass spectrometry (GCMS) and thin layer chromatography (HPTLC). Distributions of sebum components were investigated with multivariate ANOVA-simultaneous component analysis (ASCA). Sebum incretion in acne was paralleled by significantly higher abundance of triglycerides, wax esters, and squalene together with monounsaturated FAs (MUFAs), and straight chain saturated FAs (SFAs), especially those with odd-carbon chain, i.e. C13:0, C15:0, and C17:0. Profiling weight/weight percentage of individual components revealed that, in acne, the free FAs (FFAs) array was shifted towards higher relative abundance of the SFAs C15:0, C16:0, and C17:0 and lower percentage of the anteiso-branched FFAs with 12, 14, 16, and 18 carbons. In acne patients, MUFAs and PUFAs were quantitatively increased and decreased on foreheads and cheeks, respectively. Relative abundance of fatty alcohols was decreased in acne independent on the site. The results indicated that acne associates with site-specific derangement of the pathways regulating the balance among odd straight-chain and branched-chain SFAs, MUFAs, which included sapienate (C16:1n-10), PUFAs, and squalene.

Microbial synthesis of wax esters

Microbial synthesis of wax esters (WE) from low-cost renewable and sustainable feedstocks is a promising path to achieve cost-effectiveness in biomanufacturing. WE are industrially high-value molecules, which are widely used for applications in chemical, pharmaceutical, and food industries. Since the natural WE resources are limited, the WE production mostly rely on chemical synthesis from rather expensive starting materials, and therefore solution are sought from development of efficient microbial cell factories. Here we report to engineer the yeast Yarrowia lipolytica and bacterium Escherichia coli to produce WE at the highest level up to date. First, the key genes encoding fatty acyl-CoA reductases and wax ester synthase from different sources were investigated, and the expression system for two different Y. lipolytica hosts were compared and optimized for enhanced WE production and the strain stability. To improve the metabolic pathway efficiency, different carbon sources including glucose, free fatty acid, soybean oil, and waste cooking oil (WCO) were compared, and the corresponding pathway engineering strategies were optimized. It was found that using a lipid substrate such as WCO to replace glucose led to a 60-fold increase in WE production. The engineered yeast was able to produce 7.6 g/L WE with a yield of 0.31 (g/g) from WCO within 120 h and the produced WE contributed to 57% of the yeast DCW. After that, E. coli BL21(DE3), with a faster growth rate than the yeast, was engineered to significantly improve the WE production rate. Optimization of the expression system and the substrate feeding strategies led to production of 3.7–4.0 g/L WE within 40 h in a 1-L bioreactor. The predominant intracellular WE produced by both Y. lipolytica and E. coli in the presence of hydrophobic substrates as sole carbon sources were C36, C34 and C32, in an order of decreasing abundance and with a large proportion being unsaturated. This work paved the way for the biomanufacturing of WE at a large scale.

Dissolved oxygen concentration as a strategy to select type and composition of bacterial storage lipids produced during oilfield produced water treatment

Oilfield produced water (PW) management and treatment represents a significant cost to oil producers. Alcanivorax borkumensis is a marine bacteria capable of converting hydrocarbons into storage lipids, currently of great interest as raw materials for biofuel or oleochemical industries. In this study, the effect of dissolved oxygen (DO) concentration on the production of storage compounds by A. borkumensis SK2 was investigated in a sequencing batch airlift reactor treating saline oilfield produced water (PW). Intracellular lipids accumulation decreased from 0.74 g/g cellular dry weight (CDW) (7–8 mg/L O2) to 0.51 g/g CDW under 2–3 mg/L O2 and 0.45 g/g CDW during alternated DO concentrations of 7–8/1–2 mg/L O2 (famine/feast stage). Triacylglycerols were the main lipids accumulated for DO of 7–8 and 2–3 mg/L O2. During alternated DO conditions, accumulation of polyhydroxyalkanoates (0.41 g/g CDW) was observed. Extracellular lipids production were mainly wax esters and not significantly affected by DO concentration. Fatty acid (FA) composition and relative proportions of saturated and unsaturated FA in the produced compounds was also affected by oxygen availability. The obtained results show that selection of DO concentration during PW treatment can be used as a strategy to direct bacterial metabolism to production of a particular compound of interest. Hydrocarbon removal efficiencies in the range of 90%–96% were achieved showing that is possible to reduce aeration costs without compromising PW treatment and concomitant production of valuable compounds.

Lipid profiling in chilled coral larvae

Coral reefs are disappearing worldwide as a result of several harmful human activities. The establishment of cryobanks can secure a future for these ecosystems. To design effective cryopreservation protocols, basic proprieties such as chilling tolerance and lipid content must be assessed. In the present study, we investigated chilling sensitivity and the effect of chilling exposure on the lipid content and composition of larvae belonging to 2 common Indo-Pacific corals: Seriatopora caliendrum and Pocillopora verrucosa. The viability of coral larvae incubated with 0.5, 1, and 2 M ethylene glycol (EG), propylene glycol (PG), dimethyl sulfoxide (Me2SO), methanol, or glycerol and kept at 5 °C for different time periods was documented. In addition, we investigated the content of cholesterol, triacylglycerol (TAG), wax ester (WE), sterol ester (SE), lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, and several fatty acid (FA) classes in coral propagules incubated with 1 M PG or EG and kept at 5 °C for 6 h. Moreover, we examined seasonal changes in the aforementioned lipid classes in coral larvae. S. caliendrum incubated with 0.5 M PG or Me2SO and chilled for 2 h exhibited a viability rate of 11 ± 11%, whereas P. verrucosa exhibited a viability rate of 22 ± 14% after being chilled for 4 h. Furthermore, the results indicated that chilling exposure did not affect the content of any investigated lipid class in either species. The higher concentration of SE in P. verrucosa compared to S. caliendrum larvae may have contributed to the different cryotolerance displayed by the 2 larval species. A year-round lipid analysis of both coral larvae species revealed trends of homeoviscous adaptation and seasonal enhancement of lipid fluxes from symbionts to the host. During winter, the cholesterol/phospholipid ratio significantly increased, and P. verrucosa larvae exhibited an averagely decrease in FA chain lengths. During spring and summer, intracellular lipid content in the form of TAGs and WEs significantly increased in both species, and the average content of Symbiodiniaceae-derived FAs increased in P. verrucosa larvae. We concluded that the low cryotolerance displayed by S. caliendrum and P. verrucosa larvae is attributable to their chilling-sensitive membrane lipid profile and the high intracellular lipid content provided by their endosymbionts.

Comparative lipidomic analysis of Chlorella stigmatophora and Hemiselmis cf. andersenii in response to nitrogen-induced changes

The current focus of algae biotechnology is the production of high-value lipids, and its improvement by employing abiotic perturbations such as nitrogen-induced changes. In the present study, the growth dynamics, nitrogen uptake, pigments, and lipid composition of Chlorellla stigmatophora and Hemiselmis cf. andersenii were studied, in response to low (LN), medium (MN) and high (HN) nitrogen supplementations. Both microalgae responded to increased nitrogen levels by increasing their nitrogen uptake rate and pigment content. However, for lipid accumulation, C. stigmatophora presented a different pattern (LN: 16.56% > MN: 11.51% > HN: 10.95%) to that of H. cf. andersenii (MN: 15.37% > HN: 13.06% > LN: 6.71%). Untargeted gas chromatography–mass spectrometry analysis allowed the visualization of the biochemical diversity of C. stigmatophora and H. cf. andersenii, as well as differences in lipid regulation upon nitrogen-induced changes among species. For instance, glycosyl sterols were only detected for C. stigmatophora samples grown under MN and HN conditions. Moreover, lipid analysis of H. cf. andersenii, before and after alkaline hydrolysis, suggests that wax esters play a key role in the response of this microalga to high nitrogen levels. The cultivation of H. cf. andersenii at MN and HN was shown to be ideal for providing a rich source of ω3 and polyunsaturated fatty acids for nutraceutical purposes. The hierarchical cluster analysis showed the differential intra- and interspecific effects of nitrogen on lipid composition. The diverse ways by which both microalgae responded to nitrogen-induced changes highlighted the influence of phylogeny on the carbon flux through metabolic networks, and accumulation.

Effects of Microencapsulated Organic Acid and Their Salts on Growth Performance, Immunity, and Disease Resistance of Pacific White Shrimp Litopenaeus vannamei

Use of antibiotics and other chemicals to combat disease outbreaks has been a bottleneck for the sustainable growth of shrimp industry. Among various replacements proposed, organic acid (OA) and their salts (OS) are commonly used by farmers and feed millers. However, in free forms, their requirement is very high (2–3 kg/MT) as they tend to disassociate before reaching the hindgut. The dosage can be reduced by microencapsulation of the ingredients. In this study, a 63-day trial was conducted to assess the effects of OA and OS (COMP) microencapsulated (ENCAP) with fat (HF), fat + alginate (HA), wax esters (WE) and HA + WE (HAWE) on performance, digestive enzymes, immunity and resistance to Vibrio parahaemolyticus. A positive control (PC, 200 g/kg fishmeal-FM) and a negative control (NC, 130 g/kg FM) diet were formulated. Eight other diets were formulated, supplementing an NC diet with microencapsulated OA (OAHF, OAHA, OAWE, OAHAWE) and OS (OSHF, OSHA, OSWE, OSHAWE). Among the ENCAPs, significant difference was observed in serum malondialdehyde (p = 0.026), where HF showed the lowest level (6.4 ± 0.3 mmol/L). Significant interactions between COMP and ENCAP were observed in lipid deposition (p = 0.047), serum alkaline phosphatase, acid phosphatase, hepatopancreatic and serum phenol oxidase (p < 0.0001). Despite no differences, 96-h mortality during pathogenic Vibrio parahaemolyticus challenge in all treatment diets (45–56%) was lower compared to the NC diets (63%). In conclusion, use of HF microencapsulated OA diets could provide improved performance and disease resistance that could contribute to the reduction of antibiotic use by the shrimp industry.

Evolution and Characterization of Acetyl Coenzyme A: Diacylglycerol Acyltransferase Genes in Cotton Identify the Roles of GhDGAT3D in Oil Biosynthesis and Fatty Acid Composition

Cottonseed oil is rich in unsaturated fatty acids (UFAs) and serves as an edible oil in human nutrition. Reports suggest that acyl-coenzyme A: diacylglycerol acyltransferases (DGAT) and wax ester synthase/DGAT (WSD1) genes encode a key group of enzymes that catalyze the final step for triacylglycerol biosynthesis and enable an important rate-limiting process. However, their roles in oil biosynthesis and the fatty acid profile of cotton seed are poorly understood. Therefore, the aim of this study was to identify and characterize DGAT and WSD1 genes in cotton plants and examine their roles in oil biosynthesis, the fatty acid profile of cotton seeds, and abiotic stress responses. In this study, 36 GhDGAT and GhWSD1 genes were identified in upland cotton (G. hirsutum) and found to be clustered into four groups: GhDGAT1, GhDGAT2, GhDGAT3, and GhWSD1. Gene structure and domain analyses showed that the GhDGAT and GhWSD1 genes in each group are highly conserved. Gene synteny analysis indicated that segmental and tandem duplication events occurred frequently during cotton evolution. Expression analysis revealed that GhDGAT and GhWSD1 genes function widely in cotton development and stress responses; moreover, several environmental stress and hormone response-related cis-elements were detected in the GhDGAT and GhWSD1 promoter regions. The predicted target transcription factors and miRNAs imply an extensive role of GhDGAT and GhWSD1 genes in stress responses. Increases in GhDGAT3 gene expression with increases in cottonseed oil accumulation were observed. Transformation study results showed that there was an increase in C18:1 content and a decrease in C18:2 and C18:3 contents in seeds of Arabidopsis transgenic plants overexpressing GhDGAT3D compared with that of control plants. Overall, these findings contributed to the understanding of the functions of GhDGAT and GhWSD1 genes in upland cotton, providing basic information for further research.

Characterization of Oleogels Based on Waxes and Their Hydrolyzates

AbstractIn this paper, the structuring of liquid oils, also known as oleogelation, is systematically investigated for the first time using a quasi‐quaternary mixing system approach. Native waxes with different quantities of wax esters (WE), n‐alkanes (hydrocarbons (HC)), fatty acids (FA), and fatty alcohols (FaOH) are applied in mixtures with hydrolyzed waxes to systematically change the composition. Hydrolyzed waxes contain high levels of FA and FaOH. The model systems are investigated on microscopic level (brightfield light microscopy (BFM), cryogenic scanning electron microscopy (cryo‐SEM)) as well as on their macroscopic properties (rheology, gel hardness) and calorimetric behavior (differential scanning calorimetry (DSC)). It is found that sunflower wax (SFW)‐based gels (12% structurant) become less hard on any admixture. Beeswax (BW)‐based gels show significant increases in hardness when 25% and 50% (w/w) hydrolyzate are admixed. This could be related to stepwise crystallization. Further analysis reveals that the dissolution/melting behavior of the wax ester mixtures can be surprisingly well described as ideal solubility of a single pseudocomponent. The approach to unravel the individual contributions of the different species present in waxes is successful and marks a first step to better understand the systematic of wax functionality as oleogelators.Practical Application: The substitution of hardstock fats in structured oil phases is of interest for two reasons. The improved nutritional profile oleogels offer are beneficial for public health while the elimination of palm oil based ingredients appears to be a general public desire. Among the technical solutions for non‐TAG oil structuring waxes are very promising. This is primarily due to their availability, prior consumption, potentially low cost for functionality. Currently waxes are technically and scientifically wrongly treated as single components. In order to better utilize the potential of waxes and design future sourcing strategies it is necessary to understand the wax functionality at a compositional/molecular level. This contribution marks the first step into this direction by considering classes of molecules with respect to their contribution to functionality. This understanding is considered as a key for future compositional design.

The Application of a Rotating Packed Bed Reactor for the Synthesis of High Viscosity Wax Ester

Wax esters are high value-added products and widely used in a broad range of commercial fields. The enzymatic synthesis of wax ester from plant oil is more attractive than the traditional chemical method due to being environmental-friendly and limiting the use of hazardous chemicals. However, the high mass transfer resistance from the high viscous substrate leads to a low conversion and long reaction time in the continues stirred tank reactor (CSTR). The rotating packed bed reactor (RPBR) offers high mass transfer and can be used to enhance the enzymatic wax ester synthesis process. From the results, it could be concluded that the mass transfer was enhanced with the optimization of the centrifugal factor of the RPBR. Under the optimal process conditions, the wax ester yield of 96.4% was obtained after 4 hours reaction, and the half-life of the catalyst corresponded with 64 hours, while nine batches achieved of yields above 90%. The investigation proved that the RPBR is an attractive and effective reactor for heterogeneous bio-catalysis in high viscosity of 7.39~12.27 Cst at 40 °C.

Evolutionary and biochemical characterization of a Chromochloris zofingiensis MBOAT with wax synthase and diacylglycerol acyltransferase activity.

Wax synthase (WS) catalyzes the last step in wax ester biosynthesis in green plants. Two unrelated sub-families of WS, including the bifunctional acyltransferase and plant-like WS have been reported, but the latter is largely uncharacterized in microalgae. Here, we functionally characterized a putative plant-like WS (CzWS1) from the emerging model green microalga Chromochloris zofingiensis. Our results showed that plant-like WS evolved under different selection constraints in plants and microalgae, with positive selection likely contributing to functional divergence. Unlike jojoba with high amounts of wax ester in seeds and a highly active WS enzyme, C. zofingiensis has no detectable wax ester but a high abundance of WS transcripts. Co-expression analysis showed that C. zofingiensis WS has different expression correlation with lipid biosynthetic genes from jojoba, and may have a divergent function. In vitro characterization indicated that CzWS1 had diacylglycerol acyltransferase activity along with WS activity, and overexpression of CzWS1 in yeast and Chlamydomonas reinhardtii affected triacylglycerol accumulation. Moreover, biochemical and bioinformatic analyses revealed the relevance of the C-terminal region of CzWS1 in enzyme function. Taken together, our results indicated a functional divergence of plant-like WS in plants and microalgae, and the importance of its C-terminal region in specialization of enzyme function.

Jojoba Oil: An Updated Comprehensive Review on Chemistry, Pharmaceutical Uses, and Toxicity.

Jojoba is a widely used medicinal plant that is cultivated worldwide. Its seeds and oil have a long history of use in folklore to treat various ailments, such as skin and scalp disorders, superficial wounds, sore throat, obesity, and cancer; for improvement of liver functions, enhancement of immunity, and promotion of hair growth. Extensive studies on Jojoba oil showed a wide range of pharmacological applications, including antioxidant, anti-acne and antipsoriasis, anti-inflammatory, antifungal, antipyretic, analgesic, antimicrobial, and anti-hyperglycemia activities. In addition, Jojoba oil is widely used in the pharmaceutical industry, especially in cosmetics for topical, transdermal, and parenteral preparations. Jojoba oil also holds value in the industry as an anti-rodent, insecticides, lubricant, surfactant, and a source for the production of bioenergy. Jojoba oil is considered among the top-ranked oils due to its wax, which constitutes about 98% (mainly wax esters, few free fatty acids, alcohols, and hydrocarbons). In addition, sterols and vitamins with few triglyceride esters, flavonoids, phenolic and cyanogenic compounds are also present. The present review represents an updated literature survey about the chemical composition of jojoba oil, its physical properties, pharmacological activities, pharmaceutical and industrial applications, and toxicity.

Engineering Acinetobacter baylyi ADP1 for mevalonate production from lignin-derived aromatic compounds

Utilization of lignin, an abundant renewable resource, is limited by its heterogenous composition and complex structure. Biological valorization of lignin provides advantages over traditional chemical processing as it occurs at ambient temperature and pressure and does not use harsh chemicals. Furthermore, the ability to biologically funnel heterogenous substrates to products eliminates the need for costly downstream processing and separation of feedstocks. However, lack of relevant metabolic networks and low tolerance to degradation products of lignin limits the application of traditional engineered model organisms. To circumvent this obstacle, we employed Acinetobacter baylyi ADP1, which natively catabolizes lignin-derived aromatic substrates through the β-ketoadipate pathway, to produce mevalonate from lignin-derived compounds. We enabled expression of the mevalonate pathway in ADP1 and validated activity in the presence of multiple lignin-derived aromatic substrates. Furthermore, by knocking out wax ester synthesis and utilizing fed-batch cultivation, we improved mevalonate titers 7.5-fold to 1014 mg/L (6.8 mM). This work establishes a foundation and provides groundwork for future efforts to engineer improved production of mevalonate and derivatives from lignin-derived aromatics using ADP1.