Biodegradation of para-nitrophenol by Citricoccus nitrophenolicus strain PNP1T at high pH

A gram-positive bacterium Citricoccus nitrophenolicus (strain PNP1^T, DSM 23311^T, CCUG 59571^T) isolated from a waste water treatment plant was capable of effectively degrading p-nitrophenol (pNP) as a source of carbon, nitrogen and energy for growth. Degradation of pNP required oxygen and resulted in the stoichiometric release of nitrite. Strain PNP1^T also degraded 4-chlorophenol, phenol and salicylate. pNP was degraded at pH values between 6.8 and 10.0 and at temperatures between 15–32 °C. pNP at concentrations up to 150 mg L^?1 were degraded during growth in media at pH ≤ 10, whereas 200 mg L^?1 was completely inhibitory to growth. When incubated in an NH₄Cl-free medium (pH 10) containing both pNP and acetate, pNP is degraded with concomitant release of nitrite which was subsequently assimilated during acetate degradation. Intact cells of strain PNP1^T suspended in NaHCO₃/Na₂CO₃ buffer were able to continuously degrade 200 mg L^?1 pNP over a 40 day period at pH 10.     

Composition of epicuticular wax layer of two species of Mandevilla (Apocynoideae,Apocynaceae) from Rio de Janeiro,Brazil

The chemical composition of epicuticular waxes of Mandevilla guanabarica and Mandevilla moricandiana was comparatively analyzed by extraction in n-hexane and chloroform. The mean wax content per unit of leaf area in the n-hexane extract was about 13–30 μg cm⁻² for M. guanabarica, containing 20–28% n-alkanes and 55–63% triterpenes; for M. mori-candiana, the mean content was 19 μg cm⁻², containing 73% n-alkanes and 14% triterpenes. In the chloroform extract, the wax yield was 40–80 μg cm⁻² for M. guanabarica, with about 9–11% n-alkanes and 75–82% triterpenes; while for M. moricandiana, the wax yield was 110 μg cm⁻², with 52% n-alkanes and 14% triterpenes. The major compounds identified were lupeol, pentacyclic triterpenes of the α- and β-amyrin class, and n-alkanes such as nonacosane, hentriacontane and tritriacontane. These results indicate that the quantitative chemical profiles of epicuticular waxes of M. guanabarica and M. moricandiana are distinct and could be used as an additional feature in taxonomic identification.     

From crude glycerol to carotenoids by using a Rhodotorula glutinis mutant

In this work eighteen red yeasts were screened for carotenoids production on glycerol containing medium. Strain C2.5t1 of Rhodotorula glutinis, that showed the highest productivity, was UV mutagenized. Mutant 400A15, that exhibited a 280 % increase in β–carotene production in respect to the parental strain, was selected. A central composite design was applied to 400A15 to optimize carotenoids and biomass productions. Regression analyses of the quadratic polynomial equations obtained (R² = 0.87 and 0.94, for carotenoids and biomass, respectively) suggest that the models are reliable and significant (P < 0.0001) in the prediction of carotenoids and biomass productions on the basis of the concentrations of crude glycerol, yeast extract and peptone. Accordingly, total carotenoids production achieved (14.07 ± 1.45 mg l^?1) under optimized growth conditions was not statistically different from the maximal predicted (14.64 ± 1.57 mg l^?1) (P < 0.05), and it was about 100 % higher than that obtained under un-optimized conditions. Therefore mutant 400A15 may represent a biocatalyst of choice for the bioconversion of crude glycerol into value-added metabolites, and a tool for the valorization of this by-product of the biodiesel industry.     

Quercetin and silver nitrate modulate organogenesis in <Emphasis Type="Italic">Carissa carandas</Emphasis> (L.)

An in vitro organogenesis protocol for Carissa carandas L. was developed using an auxin transport inhibitor (quercetin) and silver nitrate (AgNO₃), an inhibitor of ethylene action, in association with cytokinins in the culture medium. This protocol produced the maximum number of shoots from aseptic seedling-derived shoot apex explants of C. carandas. The highest rate of shoot multiplication was recorded on MS medium containing 2.0 mg L^?1 6-benzylaminopurine; 0.5 mg L^?1 kinetin, and 0.75 mg L^?1 quercetin at after 4 wk of culture. Similar results were obtained when MS medium fortified with 2.0 mg L^?1 BAP, 0.5 mg L^?1 kinetin, and 1.5 mg L^?1 AgNO₃ was used. However, successful rooting was achieved on quarter strength MS medium with 0.5 mg L^?1 indole-3-acetic acid. In this study, an inhibitor of auxin transport and ethylene action maximized shoot multiplication in medium fortified with cytokinins. The established rapid micropropagation method could be used to conserve elite genotypes of C. carandas.     

Development of a Polygonum minus cell suspension culture system and analysis of secondary metabolites enhanced by elicitation

Polygonum minus has been reported to contain valuable metabolites and to date, there is no report on using cell culture technique for metabolite production in P. minus. Naphthalene acetic acid (NAA) concentrations in the range of 2–6 mg L^?1 were used in a matrix of combinations with dichlorophenoxyacetic acid (2,4-D) concentrations in the range of 2–10 mg L^?1 as plant growth regulators (PGRs) to induce callus cultures. Media that were supplemented with 2 mg L^?1 2,4-D + 4 mg L^?1 NAA, 2 mg L^?1 2,4-D + 6 mg L^?1 NAA and 6 mg L^?1 2,4-D + 8 mg L^?1 NAA were effective for callus induction (93.3 % of the explants produced callus). To establish cell culture, the best growth was obtained from medium that was supplemented with 1 mg L^?1 2,4-D + 2 mg L^?1 NAA. From a 1-g inoculum size, the fresh weight increases exponentially after 5–10 days of culture, and a 26.71 g maximum fresh weight was obtained after 25 days of culture. The cell culture medium was then analyzed using gas chromatography–mass spectrometry (GC–MS). Jasmonic acid (100, 50, 25 and 5 μM), salicylic acid (100, 50, 25 and 5 μM), yeast extract (500, 250 and 100 mg L^?1) and glass beads were used in this research as elicitors. The cell cultures were then incubated with the different elicitors for 1, 2, 3 and 4 days. Several compounds with high peak area percentages were detected, including 2-furancarboxaldehyde, 5-hydroxymethyl, furfural, and 2-cyclopenten-1-one, 2-hydroxy. These results show the diversity of metabolites released by P. minus cell into the culture medium under control conditions.     

n-Butanol Production from Acid-Pretreated Jatropha Seed Cake by Clostridium acetobutylicum

n-Butanol fermentation using Clostridium strains suffers from low titers due to the inability of the strains to tolerate n-butanol. The current study demonstrates a process to get high titer of n-butanol in a single batch mode from the renewable feedstock jatropha seed cake by employing Clostridium acetobutylicum. Chemical mutagenesis was done for improvement of the strain for better n-butanol tolerance and production. Optimization of the parameters resulted in 13.2 g L^?1 of n-butanol in 120 h using acid-treated jatropha seed cake hydrolysate (7 %?w/v) in anaerobic sugar medium. The process was scaled up to 15 L level, yielding 18.6 g L^?1 of n-butanol in 72 h. The strain was found to be tolerant up to 30 g L^?1 n-butanol under optimized conditions. The n-butanol tolerance was accompanied by over-expression of the stress response protein, GroEL, change in fatty acid profile, and ability to accumulate rhodamine 6G in the strain. The study has a significant impact on economically producing n-butanol from biomass.     

Effect of β-lactam antibiotics on plant regeneration in carrot protoplast cultures

Protoplasts of three carrot cultivars were isolated from in vitro-grown plantlets by overnight incubation in an enzyme mixture composed of 1% (w/v) cellulase Onozuka R-10 and 0.1% (w/v) pectolyase Y-23. After cell immobilization in modified thin alginate layers, three types of β-lactam antibiotics (cefotaxime, carbenicillin, or timentin) at five different concentrations (100, 200, 300, 400, or 500 mg L^?1) were added to the culture medium. In 20-d-old cultures, a different number of cell colonies had formed and varied on average from 27 to 56% in carbenicillin- and cefotaxime-containing media, respectively. Supplementation of the culture media with antibiotics at concentrations higher than 100 mg L^?1 resulted in a decrease in plating efficiency in comparison with the controls. However, from all antibiotic treatments, except carbenicillin at concentrations of 400–500 mg L^?1, efficient plant regeneration occurred. For this reason, we believe that cefotaxime and timentin in the concentrations analyzed here may be used in complex in vitro procedures or valuable carrot cultures as a prophylactic agent for prevention against occasional contaminations.     

Comparison of growth of Tetraselmis in a tubular photobioreactor (Biocoil) and a raceway pond

Microalgae cultivation systems can be divided broadly into open ponds and closed photobioreactors. This study investigated the growth and biomass productivity of the halophilic green alga Tetraselmis sp. MUR-233, grown outdoors in paddle wheel-driven open raceway ponds and in a tubular closed photobioreactor (Biocoil) at a salinity of 7 % NaCl (w/v) between mid-March and June 2010 (austral autumn/winter). Volumetric productivity in the Biocoil averaged 67 mg ash-free dry weight (AFDW) L^?1 day^?1 when the culture was grown without CO₂ addition. This productivity was 86 % greater, although less stable, than that achieved in the open raceway pond (36 mg L^?1 day^?1) grown at the same time in the autumn period. The Tetraselmis culture in the open raceway pond could be maintained in semi-continuous culture for the whole experimental period of 3 months without an additional CO₂ supply, whereas in the Biocoil, under the same conditions, reliable semi-continuous culture was only achievable for a period of 38 days. However, stable semi-continuous culture was achieved in the Biocoil by the addition of CO₂ at a controlled pH of ~7.5. With CO₂ addition, the volumetric biomass productivity in the Biocoil was 85 mg AFDW L^?1 day^?1 which was 5.5 times higher than the productivity achieved in the open raceway pond (15 mg AFDW L^?1 day^?1) with CO₂ addition and 8 times higher compared to the productivity in the open raceway pond without CO₂ addition (11 mg AFDW L^?1 day^?1), when cultures were grown in winter. The illuminated area productivities highlight an alternative story and showed that the open raceway pond had a three times higher productivity (3,000 mg AFDW m^?2 day^?1) compared to the Biocoil (850 mg AFDW m^?2 day^?1). Although significant differences were found between treatments and cultivation systems, the overall average lipid content for Tetraselmis sp. MUR-233 was 50 % in exponential phase during semi-continuous cultivation.     

Culture medium influence on growth,fatty acid,and pigment composition of <Emphasis Type="Italic">Choricystis minor</Emphasis> var. <Emphasis Type="Italic">minor</Emphasis>: a suitable microalga for biodiesel production

The purposes of this study were to assess the influence of culture medium on biomass production, fatty acid, and pigment composition of Choricystis minor var. minor and to evaluate the use of this microalga as a source of fatty raw material for biodiesel production. Cultures of C. minor var. minor were grown using WC (Wright’s cryptophyte) and BBM (Bold’s Basal medium) media. BBM medium produced more biomass (984.3 mg L^?1) compared to the WC medium (525.7 mg L^?1). Despite this result, WC medium produced a higher methyl ester yield for biodiesel production than the BBM medium (170.0 and 90.2 mg g^?1 of biomass, respectively). The average percentage of fatty acids obtained using the WC medium (17.0 %) was similar to soybean (18.0 %) and with similar biomass fatty acid profile. However, for pigment production, carotenoids and chlorophyll concentrations were twice as high when using the BBM medium.     

Growth and turion formation of Potamogeton crispus in response to different phosphorus concentrations in water

The vegetative growth and turion formation of Potamogeton crispus, a submersed aquatic macrophyte, was investigated under a range of phosphorus (P) concentrations (0.025, 0.25, 2.5 and 25 mg P L^?1) in the ambient water free of algae, aiming to identify the responses of submersed aquatic macrophytes to nutrient enrichment, a common eutrophication problem in China and worldwide. Plant growth was not affected by different P concentrations in terms of biomass accumulation of stems and leaves. However, the contents of chlorophyll a and starch in plants decreased with increasing water P levels, whereas chlorophyll b and carotenoids declined with P level ranging from 0.025 to 2.5 mg P L^?1. The soluble sugar content decreased when water P concentration increased up to 2.5 mg L^?1. The P content in plants increased with increasing water P levels, whereas plant N content decreased and soluble protein increased when water P concentration increased over 0.25 mg L^?1, implying that P. crispus may have modified its metabolism to adapt to water P availability. When P concentration increased to 25 mg L^?1, the number and dry matter production of turions per plant decreased significantly. Meanwhile, there was a significant reduction in turion weight and the accumulations of soluble sugar and starch in turion, when water P concentration was over 0.25 mg L^?1. The results suggest that turion formation in P. crispus is sensitive to P concentration in the ambient water, and high P levels may lead to decreases in P. crispus populations due to the decline in turion production.     

Development of doubled haploids from an elite <Emphasis Type="Italic">indica</Emphasis> rice hybrid (BS6444G) using anther culture

A total of 200 doubled haploids (DHs) were generated from an elite rice hybrid, ‘BS6444G’ for which an androgenic method was developed by manipulating the physical and chemical factors. The spike pretreated at 10?°C for 7–8 days was effective for callusing and green plant regeneration. The maximum callus frequency was achieved when the anthers cultured in N6 medium supplemented with 2.0 mg L^?1 2,4-diclorophenoxyacetic acid, 0.5 mg L^?1 6-benzylaminopurine and 3% maltose. Calli induced in N6 media also showed significant green shoot regeneration in MS medium supplemented with 0.5 mg L^?1 1-napthalene acetic acid, 0.5 mg L^?1 kinetin, 1.5 mg L^?1 benzylaminopurine and 3% sucrose producing 210 green plants. Assessment of the ploidy status showed 95.71% fertile diploids and 4.2% polyploids; no haploids were observed. A total of 38 sequence-tagged microsatellite (STMS) markers proved able to discriminate a heterozygote from all the 200 DHs. The DHs grown in the field showed significant variation for their agronomic traits. Comparison of traits with control indicates homogeneity within each DH line and significant variance of traits between DH lines. Nine DH lines produce higher grain yield than the hybrid parent which suggests the possibility of exploiting hybrid vigor in indica rice through the development of DH lines of high yielding hybrids.     

Characterization of an evolved carotenoids hyper-producer of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> through bioreactor parameter optimization and Raman spectroscopy

An evolutionary engineering approach for enhancing heterologous carotenoids production in an engineered Saccharomyces cerevisiae strain was used previously to isolate several carotenoids hyper-producers from the evolved populations. β-Carotene production was characterized in the parental and one of the evolved carotenoids hyper-producers (SM14) using bench-top bioreactors to assess the impact of pH, aeration, and media composition on β-carotene production levels. The results show that with maintaining a low pH and increasing the carbon-to-nitrogen ratio (C:N) from 8.8 to 50 in standard YNB medium, a higher β-carotene production level at 25.52 ± 2.15 mg β-carotene g^?1 (dry cell weight) in the carotenoids hyper-producer was obtained. The increase in C:N ratio also significantly increased carotenoids production in the parental strain by 298 % [from 5.68 ± 1.24 to 22.58 ± 0.11 mg β-carotene g^?1 (dcw)]. In this study, it was shown that Raman spectroscopy is capable of monitoring β-carotene production in these cultures. Raman spectroscopy is adaptable to large-scale fermentations and can give results in near real-time. Furthermore, we found that Raman spectroscopy was also able to measure the relative lipid compositions and protein content of the parental and SM14 strains at two different C:N ratios in the bioreactor. The Raman analysis showed a higher total fatty acid content in the SM14 compared with the parental strain and that an increased C:N ratio resulted in significant increase in total fatty acid content of both strains. The data suggest a positive correlation between the yield of β-carotene per biomass and total fatty acid content of the cell.     

Biodegradation of 4-bromophenol by Arthrobacter chlorophenolicus A6 in batch shake flasks and in a continuously operated packed bed reactor

The present study investigated growth and biodegradation of 4-bromophenol (4-BP) by Arthrobacter chlorophenolicus A6 in batch shake flasks as well as in a continuously operated packed bed reactor (PBR). Batch growth kinetics of A. chlorophenolicus A6 in presence of 4-BP followed substrate inhibition kinetics with the estimated biokinetic parameters value of μ _max = 0.246 h^?1, K _i = 111 mg L^?1, K _s  = 30.77 mg L^?1 and K = 100 mg L^?1. In addition, variations in the observed and theoretical biomass yield coefficient and maintenance energy of the culture were investigated at different initial 4-BP concentration. Results indicates that the toxicity tolerance and the biomass yield of A. chlorophenolicus A6 towards 4-BP was found to be poor as the organism utilized the substrate mainly for its metabolic maintenance energy. Further, 4-BP biodegradation performance by the microorganism was evaluated in a continuously operated PBR by varying the influent concentration and hydraulic retention time in the ranges 400–1,200 mg L^?1 and 24–7.5 h, respectively. Complete removal of 4-BP was achieved in the PBR up to a loading rate of 2,276 mg L^?1 day^?1.     

Agrobacterium tumefaciens-mediated transformation of bush monkey-flower (Mimulus aurantiacus Curtis) with a new reporter gene ZsGreen

A successful in vitro Agrobacterium-mediated transformation protocol was developed for Mimulus aurantiacus, a model species for ecological and evolutionary genetics and a promising ornamental plant. Three binary vectors were tested, each containing the hptII selectable marker gene and one of the reporter genes: gusA, EGFP or ZsGreen, all of them under CaMV 35S promoter. Genetic transformation was achieved through 4 days of co-cultivation of leaf, petiole and hypocotyl explants with Agrobacterium tumefaciens strain LBA 4404. Explants produced transformed callus tissue on solid modified Murashige and Skoog medium supplemented with 1 mg L^?1 6-benzylaminopurine, 0.5 mg L^?1 1-naphthaleneacetic acid, 30 g L^?1 sucrose and 20 or 50 mg L^?1 hygromycin B. All three reporter genes were expressed in callus tissue but the intensity of expression gradually decreased during further plant development. The new reporter gene ZsGreen proved suitable for plant transformation experiments since very intense and bright fluorescence was detected. Out of 1,760 co-cultured explants, 110 plants were regenerated and all of them were found to be PCR positive for the selection and/or reporter genes. Chemiluminescent Southern blot analysis revealed that 91 % of the regenerated plants (100 T₀ plants) contained T-DNA integrated in their genome. Transformation efficiency varied from 1.4 to 23.3 % for hypocotyl and petiole explants, respectively. Integration of some backbone sequences in plant genomes was confirmed in 75.3 % of T₀ plants. Using this protocol, stable transformants expressing selectable marker gene hptII and one of the reporter genes (gusA, ZsGreen or EGFP) were obtained in 4–5 months.     

Micropropagation of <Emphasis Type="Italic">Glossonema varians</Emphasis> (Stocks) Benth. ex Hook.f.—a rare Asclepiadeae of Indian Thar Desert

In the present study, an in vitro regeneration protocol for Glossonema varians (Stocks) Benth. ex Hook.f. of family Asclepiadaceae was optimized. Cotyledonary nodes of in vitro cultured seedlings were used as explants for activation of axillary shoot buds. Axillary shoot buds were initially activated on 0.1 mg L^?1 6-benzylaminopurine (BAP) and then multiplied on 0.05 mg L^?1 BAP. Shoots were rooted in vitro on 1/4 strength Murashige and Skoog medium containing 0.1 mg L^?1 2-naphthoxyacetic acid and 100 mg L^?1 activated charcoal. The cultures were maintained in a 12 h photoperiod at 40–50 μmol m^?2 s^?1 spectral flux photon, 25–30?±?2°C, and 60% relative humidity (RH). Up to 80% of in vitro regenerated plantlets were acclimatized on soilrite in cotton-plugged culture tubes in the greenhouse. This protocol can be a useful method to mass propagate and conserve this rare plant to balance biodiversity in the desert ecosystem.     

The system modulating ROS content in germinating seeds of two Brazilian savanna tree species exposed to As and Zn

The effects of increasing arsenic (0, 10, 50, 100 mg L^?1) and zinc (0, 50, 80, 120, 200 mg L^?1) doses on germination and oxidative stress markers (H₂O₂, MDA, SOD, CAT, APX, and GR) were examined in two Brazilian savanna tree species (Anadenanthera peregrina and Myracrodruon urundeuva) commonly used to remediate contaminated soils. The deleterious effects of As and Zn on seed germination were due to As- and Zn-induced H₂O₂ accumulation and inhibition of APX and GR activities, which lead to oxidative damage by lipid peroxidation. SOD and CAT did not show any As- and Zn-induced inhibition of their activities as was seen with APX and GR. We investigated the close relationships between seed germination success under As and Zn stress in terms of GR and, especially, APX activities. Increased germination of A. peregrina seeds exposed to 50 mg L^?1 of Zn was related to increased APX activity, and germination in the presence of As (10 mg L^?1) was observed only in M. urundeuva seeds that demonstrated increased APX activity. All the treatments for both species in which germination decreased or was inhibited showed decreases in APX activity. A. peregrina seeds showed higher Zn-tolerance than M. urundeuva, while the reverse was observed with arsenic up to exposures of 10 mg L^?1.     

Carbon dioxide and poultry waste utilization for production of polyhydroxyalkanoate biopolymers by <Emphasis Type="Italic">Nostoc muscorum</Emphasis> Agardh: a sustainable approach

The new paradigm is to view wastes as resources for sustainable development. In this regard, the feasibility of poultry waste and CO₂ utilization for cultivation of a filamentous nitrogen-fixing cyanobacterium, Nostoc muscorum Agardh, was investigated for production polyhydroxyalkanoates, the biodegradable polymers. This cyanobacterium showed profound rise in biomass yield with up to 10 % CO₂ supply in airstream with an aeration rate of 0.1 vvm. Maximum biomass yield of 1.12 g L^?1 was recorded for 8 days incubation period, thus demonstrating a CO₂ biofixation rate of 0.263 g L^?1 day^?1 at 10 % (v/v) CO₂-enriched air. Poultry litter (PL) supplementation also had a positive impact on the biomass yield. The nutrient removal efficiency of N. muscorum was reflected in the significant reduction in nutrient load of PL over the experimental period. A maximum poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) [P(3HB-co-3HV)] copolymer yield of 774 mg L^?1 (65 % of dry cell wt.), the value almost 11-fold higher than the control, was recorded in 10 g L^?1 PL-supplemented cultures with 10 % CO₂ supply under the optimized condition, thus demonstrating that N. muscorum has good potential for CO₂ biomitigation and poultry waste remediation while simultaneously producing eco-friendly polymers.     

Tropical Plant Extracts as Sources of Grain-Protectant Compounds Against Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae)

The toxicity of organic plant extracts to Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) was assessed for three tropical plant species: branches, leaves, and seeds of Annona montana (Annonaceae), branches of Aristolochia paulistana (Aristolochiaceae), and leaves and branches of Casearia sylvestris (Salicaceae). The screening assay resulted that the extracts of A. montana seeds obtained with hexane (LC₅₀?=?534.75 mg kg^?1 and LT₅₀?=?6.10 days) and with dichloromethane (LC₅₀?=?424.67 mg kg^?1 and LT₅₀?=?5.03 days) were the most promising treatments, followed by the extract prepared from A. montana leaves with hexane (LC₅₀?=?837.70 mg kg^?1 and LT₅₀?=?4.90 days). Moreover, extracts (at 1,500 mg kg^?1) prepared from C. sylvestris branches with dichloromethane and A. paulistana with hexane caused significant mortality (37% and 41.5%, respectively) beyond sublethal effects on S. zeamais. Therefore, based on the biological assays, extraction yield, and evaluation of the chromatographic profile of the crude extracts by TLC, the hexane extract of A. montana seeds was selected and fractioned using liquid-liquid partitioning. The hydroalcoholic fraction caused mortality of 55.5%, significantly superior to dichloromethane fraction, which caused 35.5% of mortality. Chemical analyses (¹H NMR, HPLC, and TLC) were performed, and the results showed the presence of alkaloids and acetogenins in both active fractions, which have been associated with bioactivity. Therefore, extracts prepared from A. montana seeds (especially) is an interesting source of new compounds with promising grain-protectant properties.     

Gracilibacillus xinjiangensis sp. nov., a new member of the genus Gracilibacillus isolated from Xinjiang region, China

A Gram-positive, endospore-forming, rod-shaped bacterium, designated isolate J2^T was isolated from a soil sample from Xinjiang Uyghur Autonomous Region, China. The isolate was observed to grow at 16–46 °C and pH 6.5–8.0. Chemotaxonomic analysis showed menaquinone-7 (MK-7) to be the major isoprenoid quinone; diphosphatidylglycerol, phosphatidylglycerol, one aminophospholipid, two phosphoglycolipids and one glycolipid as the major cellular polar lipids; and anteiso-C_15:0, iso-C_15:0, anteiso-C_17:0 and C_16:0 as the major fatty acids. Comparative analyses of the 16S rRNA gene sequence showed that strain J2^T is most closely related to Gracilibacillus ureilyticus (with 98.8 % similarity), Gracilibacillus dipsosauri (97.2 %), Gracilibacillus quinghaiensis (97.1 %) and Gracilibacillus thailandensis (97.0 %). The DNA–DNA reassociation values between strain J2^T and G. ureilyticus MF38^T, G. dipsosauri DD1^T, G. quinghaiensis YIM-C229^T and G. thailandensis TP2-8^T were 29.8 ± 3.7, 23.0 ± 3.5, 15.8 ± 4.9 and 15.9 ± 5.0 %, respectively. The genomic DNA G+C content of strain J2^T was determined to be 36.5 mol%. Based on these data, strain J2^T is considered as a novel species of the genus Gracilibacillus, for which the name Gracilibacillus xinjiangensis sp. nov. is proposed. The type species is J2^T (= CGMCC 1.12449^T = JCM 18859^T).     

Pilot-scale supercritical carbon dioxide extractions for the recovery of triacylglycerols from microalgae: a practical tool for algal biofuels research

We describe the preliminary extractions from a pilot-scale supercritical carbon dioxide (SC-CO₂) extractor for the isolation of algal lipids suitable for small-scale conversion to liquid hydrocarbon fuels. Flowable oils were recovered from SC-CO₂ extractions of lyophilized Nannochloropsis granulata. The extracted oils were determined to be composed primarily of triacylglycerols (TAG) by liquid chromatography–mass spectrometry analysis. Gravimetric lipid yield was increased significantly from 15.56 to 28.45 mg g^?1 ash-free dry weight (AFDW) with an increase in temperature from 50°C to 70°C, at 35 MPa over 270 min. Varying pressure had no significant effect on lipid yield. Liquid chromatography–mass spectrometry analysis of the SC-SO₂ extracts indicated that the TAG profile remained constant regardless of extraction pressure, and analysis of fatty acid methyl esters (FAME) revealed a uniform profile throughout all extraction conditions. Our optimized gravimetric lipid yields from N. granulata (28.45 mg g^?1 AFDW) were approximately half of the yields obtained by Soxhlet extraction with hexane (57.53 mg g^?1 AFDW); however, the FAME yields were similar regardless of extraction technique (18.23 mg FAME g^?1 and 17.35 mg FAME g^?1 AFDW from SC-CO₂ extraction and hexane extraction, respectively). Further extractions with Botryococcus braunii indicated that fatty acid extraction by SC-CO₂ was as efficient as hexane extraction. These results highlight the suitability of SC-CO₂ for large-scale oil extraction of microalgae for biofuel or biojet analyses due to its selectivity for TAG extraction.