Production of Mycosporine-Like Amino Acids from <Emphasis Type="Italic">Gracilaria vermiculophylla</Emphasis> (Rhodophyta) Cultured Through One Year in an Integrated Multi-trophic Aquaculture (IMTA) System

This study evaluates the production of biomass and mycosporine-like amino acids (MAAs) throughout the year in Gracilaria vermiculophylla (Rhodophyta) collected in Ria de Aveiro (Portugal). The algae were grown in outdoor tanks in seawater with the addition of fishpond effluents under two different water flows (100 and 200 L h^?1) in an integrated multi-trophic aquaculture (IMTA) system (tanks 1200 L; 1.5 m²) and different algal densities (3, 5, and 7 kg m^?2). MAA content in IMTA seaweeds was significantly affected by the interaction of time and stocking density, but not by the water flow. The highest MAA content was observed in April (about 3.13 mg g^?1 DW) followed by May (1.79 mg g^?1 DW). Seaweed biomass productivity was higher in May (372.06 g DW m^?2 week^?1) than in April (353.40 g DW m^?2 week^?1). Four MAAs were identified by HPLC and electrospray ionization mass spectrometry (ESI-MS) in G. vermiculophylla: Porphyra-334, Shinorine, Palythine and Asterina-330. The highest levels of Porphyra-334 and Shinorine were reached from November to January and the Palythine + Asterina-330 from April to August. Taking into account the average biomass and MAA production of G. vermiculophylla growing in this IMTA system (8.56 g of MAA in 18 m² culture along 8 months; 35.5% produced in April), a total amount of 71.33 g MAA year^?1 could be produced in this system by scaling up to 100 m². MAAs could be further used as photoprotector and antioxidant compounds in cosmetic products.     

Oxygen transfer as a tool for fine-tuning recombinant protein production by <Emphasis Type="Italic">Pichia pastoris</Emphasis> under glyceraldehyde-3-phosphate dehydrogenase promoter

Effects of oxygen transfer on recombinant protein production by Pichia pastoris under glyceraldehyde-3-phosphate dehydrogenase promoter were investigated. Recombinant glucose isomerase was chosen as the model protein. Two groups of oxygen transfer strategies were applied, one of which was based on constant oxygen transfer rate where aeration rate was Q _O/V = 3 and 10 vvm, and agitation rate was N = 900 min^?1; while the other one was based on constant dissolved oxygen concentrations, C _DO = 5, 10, 15, 20 and 40 % in the fermentation broth, by using predetermined exponential glucose feeding with μ _o = 0.15 h^?1. The highest cell concentration was obtained as 44 g L^?1 at t = 9 h of the glucose fed-batch phase at C _DO = 20 % operation while the highest volumetric and specific enzyme activities were obtained as 4440 U L^?1 and 126 U g^?1 cell, respectively at C _DO = 15 % operation. Investigation of specific enzyme activities revealed that keeping C _DO at 15 % was more advantageous with an expense of relatively higher by-product formation and lower specific cell growth rate. For this strategy, the highest oxygen transfer coefficient and oxygen uptake rate were K _L a = 0.045 s^?1 and OUR = 8.91 mmol m^?3 s^?1, respectively.     

Antioxidant activity and phenolic profile in filamentous cyanobacteria: the impact of nitrogen

In the present study, ethanolic extracts of ten cyanobacterial strains cultivated under different nitrogen conditions were assessed for the phenolic content and antioxidant activity. The amount of detected phenolic compounds ranged from 14.86 to 701.69 μg g^?1 dry weight (dw) and HPLC-MS/MS analysis revealed gallic acid, chlorogenic acid, quinic acid, catechin, epicatechin, kaempferol, rutin and apiin. Only catechin, among the detected phenolics, was present in all the tested strains, while quinic acid was the most dominant compound in all the tested Nostoc strains. The results also indicated the possibility of increasing the phenolic content in cyanobacterial biomass by manipulating nitrogen conditions, such as in the case of quinic acid in Nostoc 2S7B from 70.83 to 594.43 μg g^?1 dw. The highest radical scavenging activity in DPPH assay expressed Nostoc LC1B with IC₅₀ value of 0.04?±?0.01 mg mL^?1, while Nostoc 2S3B with IC₅₀ =?9.47?±?3.61 mg mL^?1 was the least potent. Furthermore, the reducing power determined by FRAP assay ranged from 8.36?±?0.08 to 21.01?±?1.66 mg AAE g^?1, and it was significantly different among the tested genera. The Arthrospira strains exhibited the highest activity, which in the case of Arthrospira S1 was approximately twofold higher in comparison to those in nitrogen-fixing strains. In addition to this, statistical analysis has indicated that detected phenolics were not major contributor to antioxidant capacities of tested cyanobacteria. However, this study highlights cyanobacteria of the genera Nostoc, Anabaena, and Arthrospira as producers of antioxidants and phenolics with pharmacological and health-beneficial effects, i.e., quinic acid and catechin in particular.     

Enhanced citric acid production by a yeast <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> over-expressing a pyruvate carboxylase gene

In this study, after the expression of a pyruvate carboxylase gene (PYC) cloned from Meyerozyma guilliermondii in a marine-derived yeast Yarrowia lipolytica SWJ-1b, a transformant PG86 obtained had much higher PYC activity than Y. lipolytica SWJ-1b. At the same time, the PYC gene expression and citric acid (CA) production by the transformant PG86 were also greatly enhanced. When glucose concentration in the medium was 60.0 g L^?1, CA concentration formed by the transformant PG86 was 34.02 g L^?1, leading to a CA yield of 0.57 g g^?1 of glucose. During a 10-L fed-batch fermentation, the final concentration of CA was 101.0 ± 1.3 g L^?1, the yield was 0.89 g g^?1 of glucose, the productivity was 0.42 g L^?1 h^?1 and only 5.93 g L^?1 reducing sugar was left in the fermented medium within 240 h of the fed-batch fermentation. HPLC analysis showed that most of the fermentation products were CA.     

Toxicity of an Annonin-Based Commercial Bioinsecticide Against Three Primary Pest Species of Stored Products

The effects of a bioinsecticide formulation based on extract of Annona squamosa L. (Annonaceae) containing 10,000 mg L^?1 of acetogenin annonin as the main active ingredient were investigated against three primary pest species of stored grains in Brazil [maize weevil Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), Mexican bean weevil Zabrotes subfasciatus (Boheman) (Coleoptera: Chrysomelidae: Bruchinae), and cowpea weevil Callosobruchus maculatus (Fabricius) (Coleoptera: Chrysomelidae: Bruchinae)] by means of residual contact bioassays. In a concentration-dependent manner, the annonin-based commercial bioinsecticide caused significant adult mortality of C. maculatus (LC₅₀ = 6890 μL kg^?1), S. zeamais (LC₅₀ = 2781 μL kg^?1), and Z. subfasciatus (LC₅₀ = 2120 μL kg^?1) after 120 h of residual contact exposure. In addition to acute toxicity, the tested bioinsecticide also promoted a significant reduction of the number of eggs laid by females of C. maculatus (EC₅₀ = 5949.7 μL kg^?1) and Z. subfasciatus (EC₅₀ = 552.7 μL kg^?1). Moreover, the bioinsecticide significantly reduced the number of emerged insects (F₁ generation) of C. maculatus (EC₅₀ = 2763.0 μL kg^?1), S. zeamais (EC₅₀ = 1380.8 μL kg^?1), and Z. subfasciatus (EC₅₀ = 561.5 μL kg^?1). The bioinsecticide also reduced the percentage of damaged grains for the three pest species studied, and its grain-protectant properties are comparable to or superior in efficacy in relation to a diatomaceous earth-based insecticide (Insecto® at 1000 mg kg^?1) used as a positive control. Thus, this standardized formulation has promising bioactivity against stored insect species and can be a useful component for IPM of stored grains in Brazil and elsewhere.     

Bioaccumulation of Trace Mercury in Trophic Levels of Benthic, Benthopelagic, Pelagic Fish Species, and Sea Birds from Arvand River, Iran

In this study, concentration of mercury was determined in the trophic levels of benthic, benthopelagic, pelagic fish species, and river birds from Arvand River, located in the Khuzestan province in the lowlands of southwestern Iran at the head of the Persian Gulf. The order of mercury concentrations in tissues of the fish species was as follows: liver>gill>muscle and in tissues of the kingfisher species was as follows: feather>liver>kidney>muscle. Therefore, liver in fish and feather in kingfisher exhibited higher mercury concentration than the other tissues. There was a positive correlation between mercury concentrations in fish and kingfisher species with size of its food items. We expected to see higher mercury levels in tissues of female species because they are larger and can eat larger food items. The results of this study show that the highest mean mercury level were found in the kingfisher (Anas crecca), followed by benthic (Epinephelus diacanthus), benthopelagic (Chanos chanos), and pelagic fish (Strongylura strongylura). Mean value of mercury in fish species, S. strongylura were (0.61 μg g^?1 dry weight), C. chanos (0.45 μg g^?1 dry weight), E. diacanthus (0.87 μg g^?1 dry weight), and in kingfisher species A. crecca was (2.64 μg g^?1 dry weight). Significant correlation between mercury concentration in fish and kingfisher may be related to high variability of mercury in the fish.     

<Emphasis Type="Italic">In vitro</Emphasis> sucrose concentration influences microtuber production and diosgenin content in white yam (<Emphasis Type="Italic">Dioscorea rotundata</Emphasis> Poir)

Dioscorea spp. is an important food crop in many countries and the source of the phytochemical diosgenin. Efficient microtuber production could provide source materials for farm-planting stock, for food markets, and for the production of high-diosgenin-producing cultivars. The first step in this study was optimizing the plant growth regulators for plantlet production, followed by a study of the effects of sucrose concentration on microtuber induction and diosgenin production. Significantly, more shoots (3.5) were produced at 4.65 μM (1 mg L^?1) kinetin (KIN), longer shoots (4.1 cm) were obtained at 2.46 μM (0.5 mg L^?1) indole-3-butyric acid (IBA), and root number (3.9) was significantly higher at 5.38 μM (1 mg L^?1) naphthalene acetic acid (NAA) than in other treatments. Increased sucrose concentrations in the optimized growth medium with 4.65 μM KIN and 5.38 μM NAA had significant effects on microtuber production (p < 0.01) and diosgenin content (p < 0.05). The most microtubers (6.2) were obtained with 100 g L^?1 sucrose, while those on 80 g L^?1 sucrose were the heaviest (0.7 g) and longest (7.4 mm). Microtubers formed in medium with 80 g L^?1 sucrose had significantly higher diosgenin content (3.64% [w/w]) than those in other sucrose treatments (< 2%) and was similar to that of field-grown parent tubers (3.79%). This result indicates an important role for sucrose in both microtuber growth and diosgenin production. Medium containing 4.65 μM KIN and 5.38 μM NAA is recommended for plantlet production, and medium containing 80 g L^?1 sucrose is recommended for microtuber and diosgenin production.     

Microbial lipid production: screening with yeasts grown on Brazilian molasses

Rhodotorula glutinis CCT 2182, Rhodosporidium toruloides CCT 0783, Rhodotorula minuta CCT 1751 and Lipomyces starkeyi DSM 70296 were evaluated for the conversion of sugars from Brazilian molasses into single-cell oil (SCO) feedstock for biodiesel. Pulsed fed-batch fermentations were performed in 1.65 l working volume bioreactors. The maximum specific growth rate (µ_max), lipid productivity (Pr) and cellular lipid content were, respectively, 0.23 h^?1, 0.41 g l^?1 h^?1, and 41 % for Rsp. toruloides; 0.20 h^?1, 0.27 g l^?1 h^?1, and 36 % for Rta. glutinis; 0.115 h^?1, 0.135 g l^?1 h^?1, and 27 % for Rta. minuta; and 0.11 h^?1, 0.13 g l^?1 h^?1, and 32 % for L. starkeyi. Based on their microbial lipid productivity, content, and profile, Rsp. toruloides and Rta. glutinis are promising candidates for biodiesel production from Brazilian molasses. All the oils from the yeasts were similar to the composition of plant oils (rapeseed and soybean) and could be used as raw material for biofuels, as well as in food and nutraceutical products.     

Process Engineering for High-Cell-Density Cultivation of Lipid Rich Microalgal Biomass of <Emphasis Type="Italic">Chlorella</Emphasis> sp. FC2 IITG

In the present study, process engineering strategy was applied to achieve lipid-rich biomass with high density of Chlorella sp. FC2 IITG under photoautotrophic condition. The strategy involved medium optimization, intermittent feeding of limiting nutrients, dynamic change in light intensity, and decoupling growth and lipid induction phases. Medium optimization was performed using combinations of artificial neural network or response surface methodology with genetic algorithm (ANN-GA and RSM-GA). Further, a fed-batch operation was employed to achieve high cell density with intermittent feeding of nitrate and phosphate along with stepwise increase in light intensity. Finally, mutually exclusive biomass and lipid production phases were decoupled into two-stage cultivation process: biomass generation in first stage under nutrient sufficient condition followed by lipid enrichment through nitrogen starvation. The key findings were as follows: (i) ANN-GA resulted in an increase in biomass titer of 157 % (0.95 g L^?1) in shake flask and 42.8 % (1.0 g L^?1) in bioreactor against unoptimized medium at light intensity of 20 μE m^?2 s^?1; (ii) further optimization of light intensity in bioreactor gave significantly improved biomass titer of 5.6 g L^?1 at light intensity of 250 μE m^?2 s^?1; (iii) high cell density of 13.5 g L^?1 with biomass productivity of 675 mg L^?1 day^?1 was achieved with dynamic increase in light intensity and intermittent feeding of limiting nutrients; (iv) finally, two-phase cultivation resulted in biomass titer of 17.7 g L^?1 and total lipid productivity of 313 mg L^?1 day^?1 which was highest among Chlorella sp. under photoautotrophic condition.     

Engineering rTCA pathway and C4-dicarboxylate transporter for <Emphasis Type="SmallCaps">l</Emphasis>-malic acid production

l-Malic acid is an important component of a vast array of food additives, antioxidants, disincrustants, pharmaceuticals, and cosmetics. Here, we presented a pathway optimization strategy and a transporter modification approach to reconstruct the l-malic acid biosynthesis pathway and transport system, respectively. First, pyruvate carboxylase (pyc) and malate dehydrogenase (mdh) from Aspergillus flavus and Rhizopus oryzae were combinatorially overexpressed to construct the reductive tricarboxylic acid (rTCA) pathway for l-malic acid biosynthesis. Second, the l-malic acid transporter (Spmae) from Schizosaccharomyces pombe was engineered by removing the ubiquitination motification to enhance the l-malic acid efflux system. Finally, the l-malic acid pathway was optimized by controlling gene expression levels, and the final l-malic acid concentration, yield, and productivity were up to 30.25 g L^?1, 0.30 g g^?1, and 0.32 g L^?1 h^?1 in the resulting strain W4209 with CaCO₃ as a neutralizing agent, respectively. In addition, these corresponding parameters of pyruvic acid remained at 30.75 g L^?1, 0.31 g g^?1, and 0.32 g L^?1 h^?1, respectively. The metabolic engineering strategy used here will be useful for efficient production of l-malic acid and other chemicals.     

Phytic acid dynamics during seed development and it’s composition in yellow and black Indian soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) genotypes through a modified extraction and HPLC method

Identifying or designing low phytic acid soybean demands an improved understanding of its dynamics and a need for a reliable and accurate phytate quantification method and hence an improved quantitative technique for accurate and rapid quantification of phytic acid (PA) using high-performance liquid chromatography is proposed in this paper. A rapid PA extraction method utilizing sonication of sample in 0.78 M HCl for 3 min followed by mechanical agitation and separation using strong anion exchange column in a vacuum manifold was optimized. The elution of PA was performed using a RP-C₁₈ column with an isocratic mobile phase [Acetonitrile, 35 mM formic acid and tetrabutylammonium hydroxide (4.8: 5.1: 0.1, v/v/v)]. The modified method was rapid, accurate, precise, and reproducible with relative standard deviation of 1.80 and 3.01 % (n = 10, for 1 mg ml^?1) for within and between days respectively with linearity (R² = 0.999, P < 0.05), low limit of detection (LOD = 7.8 μg ml^?1) and limit of quantification (LOQ = 31.25 μg ml^?1). PA dynamics was found increased in a linear trend from initial to later developing stages until maturity in both yellow (DS-9814) and black (DS-MM-64) genotypes. The PA content ranged from 2.38–4.72 g 100g^?1 in the 20 soybean genotypes screened and the variability in PA content was more in black genotypes (P < 0.0001). The black soybean genotypes JS 76-205 (2.38 g 100g^?1), Kalitur (2.50 g 100g^?1) and UPSL 652 (2.54 g 100g^?1), inherently rich in anthocyanin, contains the lowest PA content and hence can serve as potential genotypes with nutraceutical benefits.

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Graphical Abstract Pictorial representation of phytic acid extraction and quantification from soybean flour by the modified high performance liquid chromatography (HPLC) method.

     

Potential Outdoor Cultivation of Green Microalgae Based on Response to Changing Temperatures and by Combining with Air Temperature Occurrence

In this study, our working hypothesis was to examine whether temperature alters biomass and metabolite production by microalgae according to strain. We also addressed whether it is possible to choose a strain suitable for growing in each season of a given region. A factorial experiment revealed a significant interaction between chlorophylls a and b (Chl a and Chl b), carotenoid/Chl (a?+?b) ratio, biomass and total lipid productivity of six green microalgae (four Chlorella spp., Chlorella sorokiniana and Neochloris oleoabundans) after 15 days at four temperatures. At 39/35 °C, two Chlorella sp. strains (IPR7115 and IPR7117) showed higher total carotenoids/Chl (a?+?b) (0.578 and 0.830), respectively. N. oleoabundans had the highest Chl a (8210 μg L^?1) and Chl b (1909 μg L^?1) at 19/15 °C and highest maximum dry biomass (2900 mg L^?1), specific growth rate (0.538 day^?1) and total lipids (1003 mg L^?1) at 15/8 °C. We applied a method to infer the growth of these six green microalgae in outdoor ponds, as based on their response to changing temperatures and by combining with historical data on day/night air temperature occurrence for a given region. We conclude that the use of regionalized maps based on air temperature is a good strategy for predicting microalgal cultivation in outdoor ponds based on their features and tolerance to changing temperature.     

Promotion of shoot regeneration of <Emphasis Type="Italic">Swertia chirata</Emphasis> by biosynthesized silver nanoparticles and their involvement in ethylene interceptions and activation of antioxidant activity

A complete protocol for the in vitro induction of Eclipta alba tetraploids has been optimized to enhance the wedelolactone content, an anti-cancerous compound. The effects of different concentrations of colchicine (0, 0.01, 0.05, 0.1, 0.2 and 0.3%; w/v) along with treatment durations (12, 24, 36 and 48 h) were investigated on shoot tip (ST) and nodal segment (NS). The treated explants were then incubated on Murashige and Skoog (MS) medium having 1.5 mg L^?1 N⁶-benzylaminopurine and 0.5 mg L^?1 α-napthalene acetic acid for shoot regeneration and afterward root was induced on 1.0 mg L^?1 indole-3-acetic acid enriched ½MS medium. The tetraploids of E. alba were proficiently induced by the treatment of 0.1% colchicine for 24 h. The highest tetraploid induction efficiency was obtained from ST (30.56%) in comparison to the NS (22.22%). Analysis by spectrophotometry and flow cytometry showed that colchicine treated plants contained higher quantity of DNA than diploid plants. Cytological studies demonstrated doubled the chromosome number in tetraploids (2n?=?4x?=?44) than diploids (2n?=?2x?=?22). The ploidy level enhancement lead to alteration of other traits, like increased plant height, stem diameter, leaf size, stomatal size and chlorophyll content. As determined through high performance thin-layer chromatography, the ultimate achievement of this technique is the higher accumulation of wedelolactone in tetraploid plants (300.32 µg g^?1 dry weight) in evaluation to in vitro diploid (131.31 µg g^?1 dry weight) and in vivo diploid mother plants (93.26 µg g^?1 dry weight), thus improving the pharmaceutical value of E. alba.     

The Usefulness of Non-Toxic Plant Metabolites in the Control of Bacterial Proliferation

The effect of generally recognised as safe (GRAS) plant metabolites in regulating the growth of human pathogenic and probiotic bacteria and in the formation of biofilm was investigated. Thymol, carvacrol and eugenol showed the strongest antibacterial action against both pathogenic and probiotic microorganisms, at a subinhibitory concentration (SIC) of ≤50 μg ml^?1. Genistein, hydroquinone, p-hydroxybenzoic acid and resveratrol also showed antibacterial effects but at a wide concentration range (SIC = 50–1000 μg ml^?1). Catechin, gallic acid, protocatechuic acid and cranberry extracts were the most biologically compatible molecules (SIC ≥ 1000 μg ml^?1). Regarding the effect on biofilm, it was observed that thymol, carvacrol and eugenol showed antibiofilm activity against all potential pathogenic bacteria tested whilst specifically enhancing probiotic aggregation. Catechin, genistein and cranberry extracts did not inhibit the pathogenic aggregation but they stimulated probiotic biofilm formation, whilst gallic acid, protocateuchic acid, hydroquinone, p-hydroxybenzoic acid and resveratrol did not show opposite effect on biofilm formation between pathogenic and probiotic microorganisms. These results indicate that an appropriate combination of GRAS plant metabolites, which have traditionally been used as dietary constituents due to their health-promoting characteristics, can also be extremely useful in the regulation of bacterial proliferation in the intestinal microbiota. Hence, it is suggested to apply these natural GRAS molecules as dietary supplements in the food industry in order to promote probiotic viability and to prevent or reduce colonisation or proliferation of intestinal pathogens.     

Fine Root Morphology,Biochemistry and Litter Quality Indices of Fast- and Slow-growing Woody Species in Ethiopian Highland Forest

Fine root turnover of trees is a major C input to soil. However, the quality of litter input is influenced by root morphological traits and tissue chemical composition. In this study, fine roots of ten tropical woody species were collected from an Afromontane forest in the northern highlands of Ethiopia. The fine roots were analysed for root morphological traits and tissue chemistry measured as proxy carbon fractionations. Based on stem increment, the 10 species were divided into faster- and slower-growing species. Faster-growing species exhibited higher specific root length (1362 cm g^?1) than slower-growing species (923 cm g^?1). Similarly specific root area was higher in faster-growing species (223 cm² g^?1) than in slower-growing species (167 cm² g^?1). Among the carbon fractions, the acid-insoluble fraction (AIF) was the highest (44–51%). The carbon content, AIF, and the lignocellulose index were higher for slower-growing species. Root tissue density was lower in faster-growing species (0.33 g cm^?3) than slower-growing species (0.40 g cm^?3) and showed a strong positive correlation with carbon content (r ² = 0.84) and the AIF (r _pearson = 0.93). The morphological traits of fine roots between faster- and slower-growing species reflect the ecological strategy they employ. Slower-growing species have a higher tissue density which may reflect a greater longevity.     

Key parameters for outdoor biomass production of Scenedesmus obliquus in solar tracked photobioreactors

The biomass productivity of Scenedesmus obliquus was investigated outdoors during all seasons in solar tracked flat panel photobioreactors (PBR) to evaluate key parameters for process optimization. CO₂ was supplied by flue gas from an attached combined block heat and power plant. Waste heat from the power plant was used to heat the culture during winter. The parameters pH, CO₂, and inorganic salt concentrations were automatically adjusted to nonlimiting levels. The optimum biomass concentration increased directly with the photosynthetic active radiation (PAR) from 3 to 5 g dry weight (DW)?L^?1 for a low PAR of 10 mol photons m^?2 day^?1 and high PAR of 40–60 mol photons m^?2 day^?1, respectively. The annual average biomass yield (photosynthetic efficiency) was 0.4?±?0.5 g DW mol^?1 photons. However, biomass yields of 1.5 g DW mol^?1 photons close to the theoretical maximum were obtained at low PAR. The productivity (including the night biomass losses) ranged during all seasons from ?5 up to 30 g DW m^?2 day^?1 with a mean productivity of 9?±?7 g DW m^?2 day^?1. Low night temperatures of the culture medium and elevated day temperatures to the species-specific optimum increased the productivity. Thus, continuous regulation of the biomass concentration and the culture temperature with regard to the fluctuating weather conditions is essential for process optimization of outdoor microalgal production systems in temperate climates.     

Continuous succinic acid production from xylose by <Emphasis Type="Italic">Actinobacillus succinogenes</Emphasis>

Continuous, anaerobic fermentations of D-xylose were performed by Actinobacillus succinogenes 130Z in a custom, biofilm reactor at dilution rates of 0.05, 0.10 and 0.30 h^?1. Succinic acid yields on xylose (0.55–0.68 g g^?1), titres (10.9–29.4 g L^?1) and productivities (1.5–3.4 g L^?1 h^?1) were lower than those of a previous study on glucose, but product ratios (succinic acid/acetic acid = 3.0–5.0 g g^?1) and carbohydrate consumption rates were similar. Also, mass balance closures on xylose were up to 18.2 % lower than those on glucose. A modified HPLC method revealed pyruvic acid excretion at appreciable concentrations (1.2–1.9 g L^?1) which improved the mass balance closure by up to 16.8 %. Furthermore, redox balances based on the accounted xylose consumed and the excreted metabolites, indicated an overproduction of reducing power. The oxidative pentose phosphate pathway was shown to be a plausible source of the additional reducing power.     

Sweet Sorghum Juice and Bagasse as Feedstocks for the Production of Optically Pure Lactic Acid by Native and Engineered <Emphasis Type="Italic">Bacillus coagulans</Emphasis> Strains

Sweet sorghum is a bioenergy crop that produces large amounts of soluble sugars in its stems (3–7 Mg ha^?1) and generates significant amounts of bagasse (15–20 Mg ha^?1) as a lignocellulosic feedstock. These sugars can be fermented not only to biofuels but also to bio-based chemicals. The market potential of the latter may be higher given the current prices of petroleum and natural gas. The yield and rate of production of optically pure d-(?)- and l-(+)-lactic acid as precursors for the biodegradable plastic polylactide was optimized for two thermotolerant Bacillus coagulans strains. Strain 36D1 fermented the sugars in unsterilized sweet sorghum juice at 50 °C to l-(+)-lactic acid (～150 g L^?1; productivity, 7.2 g L^?1 h^?1). B. coagulans strain QZ19-2 was used to ferment sorghum juice to d-(?)-lactic acid (～125 g L^?1; productivity, 5 g L^?1 h^?1). Carbohydrates in the sorghum bagasse were also fermented after pretreatment with 0.5 % phosphoric acid at 190 °C for 5 min. Simultaneous saccharification and co-fermentation of all the sugars (SScF) by B. coagulans resulted in a conversion of 80 % of available carbohydrates to optically pure lactic acid depending on the B. coagulans strain used as the microbial biocatalyst. Liquefaction of pretreated bagasse with cellulases before SScF (L + SScF) increased the productivity of lactic acid. These results show that B. coagulans is an effective biocatalyst for fermentation of all the sugars present in sweet sorghum juice and bagasse to optically pure lactic acid at high titer and productivity as feedstock for bio-based plastics.     

Drivers of Sediment Accumulation and Nutrient Burial in Coastal Stormwater Detention Ponds,South Carolina,USA

Stormwater detention ponds are widely utilized as control structures to manage runoff during storm events. These ponds also represent biogeochemical hotspots, where carbon (C) and nutrients can be processed and buried in sediments. This study quantified C and nutrient [nitrogen (N) and phosphorus (P)] sources and burial rates in 14 stormwater detention ponds representative of typical residential development in coastal South Carolina. Bulk sediment accumulation was directly correlated with catchment impervious surface coverage (R² = 0.90) with sediment accumulation rates ranging from 0.06 to 0.50 cm y^?1. These rates of sediment accumulation and consequent pond volume loss were lower than anticipated based on maintenance guidelines provided by the State. N-alkanes were used as biomarkers of sediment source; the derived terrestrial aquatic ratio (TAR_HC) index was strongly correlated with sediment accumulation rate (R² = 0.71) which, in conjunction with high C/N ratios (16–33), suggests that terrestrial biomass drives this sediment accumulation, with relatively minimal contributions from algal derived material. This is counter to expectations that were based on the high algal productivity generally observed in stormwater ponds and previous studies of natural lakes. Sediment C and nutrient concentrations were consistent among ponds, such that differences in burial rates were a simple function of bulk sediment accumulation rate. These burial rates (C: 8.7–161 g m^?2 y^?1, N: 0.65–6.4 g m^?2 y^?1, P: 0.238–4.13 g m^?2 y^?1) were similar to those observed in natural lake systems, but lower than those observed in reservoirs or impoundments. Though individual ponds were small in area (930–41,000 m²), they are regionally abundant and, when mean burial rates are extrapolated to the regional scale (≈ 21,000 ponds), ultimately sequester 2.0 × 10⁹ g C y^?1, 9.5 × 10⁷ g N y^?1, and 3.7 × 10⁷ g P y^?1 in the coastal region of South Carolina alone. Stormwater ponds represent a relatively new but increasingly significant feature of the coastal landscape and, thus, are a key component in understanding how urbanization alters the transport and transformations of C and nutrients between terrestrial uplands and downstream receiving waters.     

In vitro propagation of <Emphasis Type="Italic">Psoralea corylifolia</Emphasis> L. by somatic embryogenesis in cell suspension culture

An effective protocol was developed for in vitro propagation of Psoralea corylifolia via somatic embryogenesis in cell suspension culture. Embryogenic callus was obtained on Murashige and Skoog (MS) medium supplemented with 6 μM naphthaleneacetic acid (NAA) and 30 μM glutamine from transverse TCLs from 10-day-old hypocotyl explants with a 96.4% frequency. Embryogenic callus produced a higher number of somatic embryos (123.7 ± 1.24 per gram fresh weight callus) on MS medium containing 30 g l^?1 sucrose, 1 μM NAA, 4 μM benzyladenine (BA), 15 μM glutamine and 2 μM abscisic acid (ABA) after 4 weeks of culture. Somatic embryos successfully germinated (97.6%) on ½ MS medium containing 20 g l^?1 sucrose, 8 g l^?1 agar and supplemented with 2 μM BA, 1 μM ABA and 2 μM gibberellic acid (GA₃) within 2 weeks of culture. Somatic embryos developed into normal plants, which hardened with 100% efficiency in soil in a growth chamber. Plants were successfully transferred to greenhouse and subsequently established in the field. Plant survival percentage in the field differed with seasonal variations. Average psoralen content of 12.9 μg g^?1 DW was measured in different stages of somatic embryo development by high-performance liquid chromatography (HPLC). This protocol will be helpful for efficient propagation of elite clones on a mass scale, conservation efforts of this species and for secondary metabolites production studies.