Scale-up of microbial lipid and bioethanol production from oilcane.

Microbial oils are a sustainable biomass-derived substitute for liquid fuels and vegetable oils. Oilcane, an engineered sugarcane with superior feedstock characteristics for biodiesel production, is a promising candidate for bioconversion. This study describes the processing of oilcane stems into juice and hydrothermally pretreated lignocellulosic hydrolysate and their valorization to ethanol and microbial oil using Saccharomyces cerevisiae and engineered Rhodosporidium toruloides strains, respectively. A bioethanol titer of 106 g/L was obtained from S. cerevisiae grown on oilcane juice in a 3 L fermenter, and a lipid titer of 8.8 g/L was obtained from R. toruloides grown on oilcane hydrolysate in a 75 L fermenter. Oil was extracted from the R. toruloides cells using supercritical CO2, and the observed fatty acid profile was consistent with previous studies on this strain. These results demonstrate the feasibility of pilot-scale lipid production from oilcane hydrolysate as part of an integrated bioconversion strategy.

Critical Review on the Use of Extractives of Naturally Durable Woods as Natural Wood Protectants.

Naturally durable wood pre-dates preservative-treated wood and has been demonstrated to offer a suitable service life for certain applications where preservative-treated wood is not feasible. Heartwood extractives have been demonstrated to impart bio-deteriorative resistance to naturally durable wood species. These extractives are typically found in the heartwood of living trees and are produced either by the death of parenchyma cells or as the result of external stimuli. The mechanisms of natural durability are not well understood, as heartwood extractives can be extremely variable in their distribution, composition, and efficacy in both living and harvested trees. The underlying complexity of heartwood extractives has hindered their standardization in residential building codes for use as wood preservatives. The use of naturally durable lumber is not always feasible, as woods with exceptionally durable heartwood do not typically yield lumber with acceptable machining properties. A potential approach to overcome the inherent difficulty in establishing guidelines for the appropriate use of naturally durable wood is to focus solely on the extractives as a source of bioactive protectants based on the strategies used on living and dead wood to repel the agents of biodeterioration. This critical review summarizes the relevant literature on naturally durable woods, their extractives, and their potential use as bio-inspired wood protectants. An additional discussion will be aimed at underscoring the past difficulties in adopting this approach and how to overcome the future hurdles.

In Vitro Evaluation of the Adsorption Efficacy of Biochar Materials on Aflatoxin B1, Ochratoxin A, and Zearalenone.

Mycotoxin sequestration materials are important tools to reduce mycotoxin illness and enable proper handling of mycotoxin-contaminated commodities. Three food-grade bentonite clays and four generally recognized as safe (GRAS) charcoal/biochar carbon materials that are marketed as feed additives and supplements were evaluated for their ability to sequester the mycotoxins aflatoxin B1, ochratoxin A, and zearalenone. The surface area of the clays varied between 32.1 to 51.4 mg2/g, and the surface area of the carbon-based materials varied from 1.7 to 1735 mg2/g. In vitro, gastric fluid studies indicated that certain pine biochar and activated coconut charcoal could sequester high amounts (85+%) of the mycotoxins at 1 ppm levels or below. However, some biochar materials with lower surface area properties lacked binding capacity. The coconut shell charcoal and pine biochar utilize agricultural waste products in a manner that significantly reduces carbon emissions and provides valuable materials to minimize exposure to toxins found in food and feed.

Repellency and toxicity of a CO2-derived cedarwood oil on hard tick species (Ixodidae).

The repellency and toxicity of a CO2-derived cedarwood oil (CWO) was evaluated against actively questing unfed nymphs of four species of hard ticks: Amblyomma americanum (L.), Dermacentor variabilis (Say), Ixodes scapularis Say, and Rhipicephalus sanguineus (Latreille). Using a vertical climb bioassay for repellency, nymphs of these species avoided a CWO-treated filter paper in proportional responses to treatment concentrations. At 60 min of exposure, I. scapularis nymphs were most sensitive with 50% repellency concentration (RC50) of 19.8 µg cm-2, compared with RC50 of 30.8, 83.8 and 89.6 µg cm-2 for R. sanguineus, D. variabilis and A. americanum, respectively. Bioassays determined the lethal concentration for 50% (LC50) and 90% (LC90) mortality of nymphs exposed to CWO in treated vials after 24- and 48-h exposure. After 24 h exposure, the LC50 values were 1.25, 3.45 and 1.42 µg cm-2 and LC90 values were 2.39, 7.59 and 4.14 µg cm-2 for D. variabilis, I. scapularis and R. sanguineus, respectively, but had minimal effect on A. americanum. After 48 h exposure, the LC50 values were 4.14, 0.78, 0.79 and 0.52 µg cm-2, and LC90 values were 8.06, 1.48, 1.54 and 1.22 µg cm-2 for A. americanum, D. variabilis, I. scapularis and R. sanguineus, respectively. The repellency of CWO on tick species decreased with time. The repellency and toxicity bioassays demonstrated concentration-dependent responses of tick nymphs to the oil, indicating the potential of the CO2-derived cedarwood oil be developed as an eco-friendly repellent and/or acaricide.

Liposomes Loaded with Unsaponifiable Matter from Amaranthus hypochondriacus as a Source of Squalene and Carrying Soybean Lunasin Inhibited Melanoma Cells.

Amaranthus hypochondriacus is a source of molecules with reported health benefits such as antioxidant activity and cancer prevention. The objective of this research was to optimize the conditions for preparing a liposome formulation using amaranth unsaponifiable matter as a source of squalene in order to minimize the particle size and to maximize the encapsulation efficiency of liposomes for carrying and delivering soybean lunasin into melanoma cell lines. Amaranth oil was extracted using supercritical dioxide carbon extraction (55.2 MPa pressure, 80 °C temperature, solvent (CO2)-to-feed (oil) ratio of 20). The extracted oil from amaranth was used to obtain the unsaponifiable enriched content of squalene, which was incorporated into liposomes. A Box-Behnken response surface methodology design was used to optimize the liposome formulation containing the unsaponifiable matter, once liposomes were optimized. Soybean lunasin was loaded into the liposomes and tested on A-375 and B16-F10 melanoma cells. The squalene concentration in the extracted oil was 36.64 ± 0.64 g/ 100 g of oil. The particle size in liposomes was between 115.8 and 163.1 nm; the squalene encapsulation efficiency ranged from 33.14% to 76.08%. The optimized liposome formulation contained 15.27 mg of phospholipids and 1.1 mg of unsaponifiable matter. Cell viability was affected by the liposome formulation with a half-maximum inhibitory concentration (IC50) equivalent to 225 µM in B16-F10 and 215 µM in A-375. The liposomes formulated with lunasin achieved 82.14 ± 3.34% lunasin encapsulation efficiency and improved efficacy by decreasing lunasin IC50 by 31.81% in B16-F10 and by 41.89% in A-375 compared with unencapsulated lunasin.

Utilization of Quercetin as an Oviposition Stimulant by Lab-Cultured Coleomegillamaculata in the Presence of Conspecifics and a Tissue Substrate.

BACKGROUND: The discovery of natural products to improve the reproductive performance of mass-reared predators is an important aim for successful augmentative biological control. We tested the hypothesis that quercetin (a bioflavonoid) stimulates oviposition by the ladybird beetle Coleomegillamaculata in the presence of conspecifics and a tissue substrate. METHODS: We conducted bioassays in solitary cages (housing one female) and communal cages (housing 10 females) to estimate daily oviposition site preferences, egg production in response to quercetin in the presence or absence of a tissue paper substrate, and female “resting” positions. RESULTS: Females preferentially oviposited within 1⁻2 cm of quercetin powder, held in a tiny dish, at the base of cages. When given a choice, females oviposited in the dish with quercetin over a tissue paper substrate. In one of two experiments, they produced more egg clutches, regardless of oviposition site, when the quercetin and tissue were in close juxtaposition. Females “rested” on the tissue in the presence or absence of quercetin. CONCLUSION: This study provides evidence that quercetin can be utilized as an oviposition stimulant by C. maculata in a rearing system. Future research should determine if quercetin stimulates oviposition in other ladybird beetle species.

Do Bioflavonoids in Juniperus virginiana Heartwood Stimulate Oviposition in the Ladybird Coleomegilla maculata?

Maximizing the reproductive potential of ladybird beetles fed factitious foods or artificial diets, in lieu of natural prey, is a major challenge to cost-effective mass rearing for augmentative biological control. In this study, we tested the hypothesis that compounds in redcedar, Juniperus virginiana, stimulate oviposition in the ladybird Coleomegilla maculata. We also tested the prediction that several bioflavonoids, identified in heartwood fractions, elicited this behavioral response. Phenolic compounds were extracted from J. virginiana heartwood sawdust, separated into several fractions, then presented to adult beetles, in a powdered, pure form, in the laboratory. Females preferentially oviposited within 1 to 2 cm of fractions B, C, D, and E, but not A or the unfractionated extract, at the base of test cages. Chemical analysis identified bioflavonoids in heartwood fractions and subsequent bioassays using several identified in fractions C, D, and E confirmed that quercetin, taxifolin, and naringenin (to a lesser extent) stimulated oviposition. All tested fractions and bioflavonoids readily adhered to the chorion of freshly laid eggs but did not reduce egg hatch. This study demonstrates that several bioflavonoids stimulate oviposition by C. maculata and could be useful for mass rearing programs.

Recovery of Butanol by Counter-Current Carbon Dioxide Fractionation with its Potential Application to Butanol Fermentation.

A counter-current CO2 fractionation method was applied as a mean to recover n-butanol and other compounds that are typically obtained from biobutanol fermentation broth from aqueous solutions. The influence of operating variables, such as solvent-to-feed ratio, temperature, pressure and feed solution composition was experimentally studied in terms of separation efficiency, butanol removal rate, total removal and butanol concentration in the extract at the end of the continuous cycle. With respect to the temperature and pressure conditions investigated, results show that the highest separation efficiency was obtained at 35 °C and 10.34 MPa. At these operating conditions, 92.3% of the butanol present in the feed solution was extracted, and a concentration of 787.5 g·L-1 of butanol in the extract was obtained, starting from a feed solution of 20 g·L-1. Selectivity was calculated from experimental data, concluding that our column performs much better than a single equilibrium stage. When adding ethanol and acetone to the feed solution, ethanol was detected in the water-rich fraction (raffinate), whereas the highest concentration of acetone was found in the butanol rich fraction (extract).

Frost Grape Polysaccharide (FGP), an Emulsion-Forming Arabinogalactan Gum from the Stems of Native North American Grape Species Vitis riparia Michx.

A new arabinogalactan is described that is produced in large quantity from the cut stems of the North American grape species Vitis riparia (Frost grape). The sugar composition consists of l-arabinofuranose (l-Araf, 55.2%) and d-galactopyranose (d-Galp 30.1%), with smaller components of d-xylose (11.2%), d-mannose (3.5%), and glucuronic acid (GlcA, ∼2%), the latter linked via a galactosyl residue. Permethylation identified 3-linked Galp residues, some substituted at the 2-position with Galp or Manp, terminal Araf and Xylp, and an internal 3-substituted Araf. NMR (HSQC, TOCSY, HMBC, DOSY) identified ßGalp and three αAraf spin systems, in an Araf-α1,3-Araf-α1,2-Araf-α1,2-Galp structural motif. Diffusion-ordered NMR showed that the FGP has a molecular weight of 1-10 MDa. Unlike gum arabic, the FGP does not contain a hydroxyproline-rich protein (HPRP). FGP forms stable gels at >15% w/v and at 1-12% solutions are viscous and are excellent emulsifiers of flavoring oils (grapefruit, clove, and lemongrass), giving stable emulsions for ≥72 h. Lower concentrations (0.1% w/v) were less viscous, yet still gave stable grapefruit oil/water emulsions. Hence, FGP is a ß1,3-linked arabinogalactan with potential as a gum arabic replacement in the food and beverage industries.

Antioxidant activity and sensory evaluation of a rosmarinic acid-enriched extract of Salvia officinalis.

An extract of Salvia officinalis (garden sage) was prepared using supercritical carbon dioxide (SC-CO2 ) extraction, followed by hot water extraction. The resulting extract was enriched in polyphenols, including rosmarinic acid (RA), which has shown promising health benefits in animals. The extract contained RA at a concentration of 28.4 mg/g, representing a significant enrichment from the RA content in sage leaves. This extract was incorporated into oil-in-water emulsions as a source of lipid antioxidants and compared to emulsions containing pure rosmarinic acid. Both treatments were effective in suppressing lipid oxidation. The extract was evaluated by a trained sensory panel in a tea formulation. While the panel could discriminate among extract-treated and control samples, panelists demonstrated high acceptability of the sage extract in a tea.

Factors Affecting Pheromone Production by the Pepper Weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae) and Collection Efficiency.

Several factors affecting pheromone production by male pepper weevils, Anthonomus eugenii Cano (Coleoptera: Curculionidae) as well as collection efficiency were investigated. Factors studied included: porous polymer adsorbents (Tenax versus Super Q), male age, time of day, male density, and male diet. Super Q was found to be a superior adsorbent for the male-produced alcohols and geranic acid as well as the plant-produced E-ß-ocimene. Pheromone production increased with male age up to about age 15 days old and then tapered off. Male pepper weevils produced the highest amount of pheromone between noon and 2 pm (i.e., 4 to 6 h after "lights on") and were producing ca. 800 ng/h during this period. Thereafter, pheromone production decreased and was extremely low during the scotophase (i.e., ca. 12 ng/h). Male pepper weevil density had a significant effect on both release rate and pheromone composition. Pheromone production on a per male basis was highest for individual males and the percentage of geranic acid in the blend was lowest for individual males. Male pepper weevils produced only extremely low amounts of pheromone when feeding on artificial diet; however, they produced very high amounts when on fresh peppers. Together, this information will be useful in designing better attractant lures for pepper weevils.

Topical wound-healing effects and phytochemical composition of heartwood essential oils of Juniperus virginiana L., Juniperus occidentalis Hook., and Juniperus ashei J. Buchholz.

Ethnobotanical surveys indicated that in the traditional medicines worldwide, several Juniperus species are utilized as antihelmintic, diuretic, stimulant, antiseptic, carminative, stomachic, antirheumatic, antifungal, and for wound healing. In the present study, essential oils obtained from heartwood samples of Juniperus virginiana L., Juniperus occidentalis Hook. and Juniperus ashei J. Buchholz were evaluated for wound healing and anti-inflammatory activities by using in vivo experimental methods. The essential oils were obtained by the supercritical carbon dioxide extraction method. Linear incision and circular excision wound models were performed for the wound-healing activity assessment. The tissues were also evaluated for the hydroxyproline content as well as histopathologically. To evaluate the anti-inflammatory activity of the essential oils, the test used was an acetic acid-induced increase in capillary permeability. The essential oil of J. occidentalis showed the highest activity on the in vivo biological activity models. Additionaly, the oil of J. virginiana was found highly effective in the anti-inflammatory activity method. The experimental data demonstrated that essential oil of J. occidentalis displayed significant wound-healing and anti-inflammatory activities.

Phytosterol and tocopherol components in extracts of corn distiller's dried grain.

As the ethanol industry continues to grow, it will become very important to develop value-added markets for its coproducts in order for the industry to remain profitable. Corn distiller's dried grain (DDG) is a major coproduct of ethanol fermentation from corn processed by dry-milling and is primarily sold as livestock feed. The objective of this research was to determine if valuable phytochemicals found in corn oil and corn fiber oil, such as phytosterols and their saturated equivalents, phytostanols, ferulate phytosterol esters (FPE), tocopherols, and tocotrienols, are retained in DDG. Hexane and supercritical carbon dioxide (CO2) extracts of DDG were similar in their concentrations of total phytosterols (15.8-17.3 mg/g of extract), FPE (3.75-3.99 mg/g of extract), and tocols (1.7-1.8 mg/g of extract). Ethanol extracts were slightly lower in concentration of phytosterols (8.9-11.4 mg/g of extract), FPE (1.62-1.98 mg/g of extract), and tocols (0.73-0.76 mg/g of extract).

Oil and tocopherol content and composition of pumpkin seed oil in 12 cultivars.

Twelve pumpkin cultivars (Cucurbita maxima D.), cultivated in Iowa, were studied for their seed oil content, fatty acid composition, and tocopherol content. Oil content ranged from 10.9 to 30.9%. Total unsaturated fatty acid content ranged from 73.1 to 80.5%. The predominant fatty acids present were linoleic, oleic, palmitic, and stearic. Significant differences were observed among the cultivars for stearic, oleic, linoleic, and gadoleic acid content of oil. Low linolenic acid levels were observed (<1%). The tocopherol content of the oils ranged from 27.1 to 75.1 microg/g of oil for alpha-tocopherol, from 74.9 to 492.8 microg/g for gamma-tocopherol, and from 35.3 to 1109.7 microg/g for delta-tocopherol. The study showed potential for pumpkin seed oil from all 12 cultivars to have high oxidative stability that would be suitable for food and industrial applications, as well as high unsaturation and tocopherol content that could potentially improve the nutrition of human diets.

Enhanced selective extraction of hexane from hexane/soybean oil mixture using binary gas mixtures of carbon dioxide.

Carbon dioxide (CO2) can effectively separate hexane from a mixture of soybean oil (SBO) and hexane with a slight coextraction of SBO. Previous research demonstrated that CO2 entrained with helium significantly reduced SBO solubility in CO2. In this study, CO2 was mixed with three gases (He, N2, or Ar) (0.5-30 vol %) to decrease SBO solubility while attempting to maintain hexane solubility. The binary gas mixtures (at 25 degrees C and 9.31 MPa) were passed through a 25 wt % hexane/SBO mixture inside a 2.5 m fractionation column. Coextracted SBO was inversely proportional to binary gas concentration, whereas residual hexane in the raffinate was proportional to binary gas concentration. The 10% binary mixture of N2 or Ar was the best compromise to obtain both low residual hexane levels (i.e., 26 ppm) and low SBO coextraction (i.e., only 40 mg). This carry-over of SBO represents a 95% reduction in SBO carry-over compared to neat CO2.

Supercritical fluid chromatographic analysis for on-line monitoring of hexane removal from soybean oil miscella using liquid carbon dioxide.

Liquid carbon dioxide (L-CO2) can be used to separate hexane from hexane/soybean oil (SBO) mixtures (i.e., miscella). An on-line supercritical fluid chromatographic (SFC) method was developed to monitor this separation. L-CO2 (25 degrees C and 9.31 MPa) was passed through 50 mL of a 25% (w/w) hexane miscella and then directed on-line through a SFC injector. After passing 300-L expanded CO2, the hexane concentrations in the L-CO2 were 0.05% and 0.04% for n-hexane and isohexane, respectively and the residual hexane concentrations in the SBO were 3.8 and 3.3 ppm, respectively. This technique provided real time on-line monitoring of the hexane separation process.

Preparation of spread oils meeting U.S. Food and Drug Administration Labeling requirements for trans fatty acids via pressure-controlled hydrogenation.

On July 11, 2003, the U.S. Food and Drug Administration (FDA) announced final regulations for trans fatty acid (TFA) labeling. By January 1, 2006, the TFA content of foods must be labeled as a separate line on the Nutrition Facts label. Products containing <0.5 g of TFA/14 g serving may be declared as zero. This paper describes technologies allowing compliance with TFA labeling requirements. Soybean oil was hydrogenated in a 2-L vessel at temperatures ranging from 120 to 170 degrees C at a hydrogen pressure of 200 psi. A commercial nickel-supported catalyst (25% Ni) was used at 0.02% Ni by weight of oil. The hydrogenated oils were characterized for fatty acid composition, solid fat content, and melting point. Compared to commercially processed soybean oil basestocks that typically contain approximately 40% TFA, those obtained at lower temperatures and higher pressures contain >56% less TFA. Basestocks prepared in the laboratory when blended with liquid soybean oil will yield spread oils meeting FDA labeling requirements for zero TFA, that is, <0.5 g of TFA/serving.

Pressurized fluids for extraction of cedarwood oil from Juniperus virginianna.

The extraction of cedarwood oil (CWO) using liquid carbon dioxide (LC-CO(2)) was investigated and compared to supercritical fluid extraction, including the effects of extraction pressure and length of extraction. The chemical composition of the extracts was monitored over the course of the extraction as well. The cumulative yields of CWO from cedarwood chips using 80 L of carbon dioxide varied very little treatment to treatment, with all temperature/pressure combinations yielding between 3.55 and 3.88% CWO, and the cumulative yields were statistically equivalent. The rate of extraction was highest under the supercritical extraction conditions (i.e., 100 degrees C and 6000 psi). Under the liquid CO(2) conditions (i.e., 25 degrees C), the extraction rates did not vary significantly with extraction pressure. However, there were differences in the chemical composition of the collected CWO. Extractions at 100 degrees C gave a much lower ratio of cedrol/cedrene than extractions at 25 degrees C. The highest ratio of cedrol/cedrene was obtained using 25 degrees C and 1500 psi. The use of subcritical water was also investigated for the extraction of CWO as well. Although some CWO was extracted using this method, the temperature/pressure combinations that gave the highest weight percentage yields also gave oils with an off odor while those combinations that gave a higher quality oil had very low yields. It appears that the high temperatures and acidic conditions cause a dehydration of the tertiary alcohol, cedrol, to its hydrocarbon analogue, cedrene, during CO(2) or pressurized water extractions of cedarwood.

Enhancing quality and oxidative stability of aged fried food with gamma-tocopherol.

To determine the effects of gamma-tocopherol on the stability of fried food, potato chips were fried in triolein with 0, 100, or 400 ppm gamma-tocopherol. Potato chips, sampled at 1, 3, and 6 h of frying time, were aged for 0, 2, and 4 days at 60 degrees C and then evaluated for odor attributes by sensory analysis and for volatile compounds by purge-and-trap gas chromatography-mass spectrometry. Oil sampled after 1, 3, and 6 h of frying time from the fryer was evaluated for total polar compounds and retention of gamma-tocopherol. Oil extracted from the potato chips was also analyzed for residual gamma-tocopherol. gamma-Tocopherol disappeared rapidly, with only slight amounts of the original 100 ppm level detectable after the triolein was used for frying. gamma-Tocopherol significantly inhibited polar compound production in the triolein. Results showed that gamma-tocopherol inhibited the oxidation of the fried food even when only very low levels of retained gamma-tocopherol were present in the frying oil or potato chips. Nonanal formation was inhibited by gamma-tocopherol in aged potato chips. Odor analysis of the aged potato chips showed that samples with 0 ppm gamma-tocopherol had a rancid odor after being aged for 4 days. Potato chips with 400 ppm gamma-tocopherol had no rancid odors; however, as the level of gamma-tocopherol decreased in the triolein and in the potato chips, a weak plastic odor characteristic of oxidized triolein was detected.