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.     

Isolation and screening of euryhaline <Emphasis Type="Italic">Tetraselmis</Emphasis> spp. suitable for large-scale outdoor culture in hypersaline media for biofuels

Natural saline lakes in Western Australia were sampled for microalgae species and strains with potential for large-scale outdoor cultivation over a wide range of salinities for biofuels production. Using a rational isolation and screening process, several Tetraselmis strains (Chlorophyta, Chlorodendrales) with a broad range of salinity tolerance were identified and were characterised further for their potential for biofuels production. Specific growth rates increased from 0.8 to 1.2 days^?1 when the medium salinity was decreased from 11 to 3 % (w/v) NaCl (1.88 to 0.51 M NaCl) in batch cultivation mode, thereby indicating quick adaptation to large salinity changes. In general, ash-free dry weight (AFDW), total lipid, protein and carbohydrate contents per cell were highest in the early stages of growth. Salinity increases led to an increase in cell AFDW, with the highest mean maximum of 2555?±?659 pg AFDW.cell^?1 at 11 % (w/v) NaCl in the strains Tetraselmis MUR 167 and MUR 219 which had been in culture for many years, as compared to the mean maximum of 981?±?141 pg AFDW.cell^?1 the in newly isolated strains MUR 230, 231, 232 and 233. Similar observations on total lipid, protein and carbohydrate content per cell were made between the two groups of strains. Overall, all strains yielded high biomass and total lipid productivities over a very wide range of salinities without large variation in their gross biochemical composition and growth pattern. Based on AFDW and total lipid productivity data, the order of preference for selecting strains for further investigation for large-scale culture was MUR 231?>?MUR 233?>?MUR 219?>?MUR 230?>?MUR 232?>?MUR 167. The Tetraselmis spp. were also very competitive as shown by the outdoor cultivation of diatom, Halamphora coffeaeformis MUR 158, in parallel with Tetraselmis sp. MUR 167 which resulted in the diatom being outcompeted by the green alga. Our results demonstrate the high commercial potential of euryhaline Tetraselmis spp. for cultivation over a broad range of salinity in outdoor cultures.     

High biomass production and CO<Subscript>2</Subscript> fixation from biogas by <Emphasis Type="Italic">Chlorella</Emphasis> and <Emphasis Type="Italic">Scenedesmus</Emphasis> microalgae using tequila vinasses as culture medium

Tequila vinasses (TVs) generated during Tequila production are brown liquid residues rich in nutrients. The nutrient content of agro-industrial effluents represents an excellent resource to support low-cost biomass production of microalgae; nonetheless, it is crucial to select the suitable microalgal strain to attain the highest biomass production in each residue used. In this study, biomass production, CO₂ fixation from biogas, and cell compound accumulation by Chlorella vulgaris U162, Chlorella sp., Scenedesmus obliquus U169, and Scenedesmus sp. using biodigested and filtered TVs as culture medium were evaluated and compared with the conventional microalgal culture media, C30, BG-11, Bold 3N, and Bristol. The four microalgae evaluated attained the highest biomass production and CO₂ fixation rate cultured in both residues, accumulating mainly carbohydrates and proteins although the most appropriate microalga to be cultured in TVs was Chlorella sp., recording 2.30 g L^?1. Moreover, the nutrient ratio of filtered TVs was ideal to support biomass production while biodigested TVs need to be supplemented with nitrogen. Overall, these results demonstrated that tequila vinasses are an excellent resource to support high and quick biomass production of microalgae, which can be used to obtain biofuels as ethanol, biogas, and supplement food depicting an extra benefit during the appropriate disposal of this residue.     

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.     

Productivity and biochemical composition of <Emphasis Type="Italic">Tetradesmus obliquus</Emphasis> and <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis>: effects of different cultivation approaches

The present work evaluated biomass productivity, carbon dioxide fixation rate, and biochemical composition of two microalgal species, Phaeodactylum tricornutum (Bacillariophyta) and Tetradesmus obliquus (Chlorophyta), cultivated indoors in high-technology photobioreactors (HT-PBR) and outdoors both in pilot ponds and low-technology photobioreactors in a greenhouse in southern Italy. Microalgae were grown in standard media, under nitrogen starvation, and in two liquid digestates obtained from anaerobic digestion of agro-zootechnical and vegetable biomass. P. tricornutum, cultivated in semi-continuous mode in indoor HT-PBRs with standard medium, showed a biomass productivity of 21.0?±?2.3 g m^?2 d^?1. Applying nitrogen starvation, the lipid productivity increased from 2.3 up to 4.5?±?0.5 g m^?2 d^?1, with a 24 % decrease of biomass productivity. For T. obliquus, a biomass productivity of 9.1?±?0.9 g m^?2 d^?1 in indoor HT-PBR was obtained using standard medium. Applying liquid digestates as fertilizers in open ponds, T. obliquus gave a biomass productivity (10.8?±?2.0 g m^?2 d^?1) not statistically different from complete medium such as P. tricornutum (6.5?±?2.2 g m^?2 d^?1). The biochemical data showed that the fatty acid composition of the microalgal biomass was affected by the different cultivation conditions for both microalgae. In conclusion, it was found that the microalgal productivity in standard medium was about doubled in HT-PBR compared to open ponds for P. tricornutum and was about 20 % higher for T. obliquus.     

<Emphasis Type="Italic">Pseudomonas</Emphasis> spp. increases root biomass and tropane alkaloid yields in transgenic hairy roots of <Emphasis Type="Italic">Datura</Emphasis> spp.

Transgenic hairy roots of Datura spp., established using strain A4 of Agrobacterium rhizogenes, are genetically stable and produce high levels of tropane alkaloids. To increase biomass and tropane alkaloid content of this plant tissue, four Pseudomonas strains, Pseudomonas fluorescens P64, P66, C7R12, and Pseudomonas putida PP01 were assayed as biotic elicitors on transgenic hairy roots of Datura stramonium, Datura tatula, and Datura innoxia. Alkaloids were extracted from dried biomass, and hyoscyamine and scopolamine were quantified using liquid chromatography-tandem mass spectrometry analysis. D. stramonium and D. innoxia biomass production was stimulated by all Pseudomonas spp. strains after a 5-d treatment. All strains of P. fluorescens increased hyoscyamine yields compared to untreated cultures after both 5 and 10 d of treatment. Hyoscyamine yields were highest in D. tatula cultures exposed to a 5-d treatment with C7R12 (16.633 + 0.456 mg g^?1 dry weight, a 431% increase) although the highest yield increases compared to the control were observed in D. stramonium cultures exposed to strains P64 (511% increase) and C7R12 (583% increase) for 10 d. D. innoxia showed the highest scopolamine yields after elicitation with P. fluorescens strains P64 for 5 d (0.653 + 0.021 mg g^?1 dry weight, a 265% increase) and P66 for 5 and 10 d (5 d, 0.754 + 0.0.031 mg g^?1 dry weight, a 321% increase; 10 d 0.634 + 0.046 mg g^?1 dry weight, a 277% increase). These results show that the Pseudomonas strains studied here can positively and significantly affect biomass and the yields of hyoscyamine and scopolamine from transgenic roots of the three Datura species.     

Energetic evaluation of an internally illuminated photobioreactor for algal cultivation

An annular internally illuminated photobioreatcor (IIPBR) configuration based on the airlift/bubble column principles was developed and validated at an 18 l prototype scale using Scenedemus sp. and Nannochloropsis salina in batch and semi-continuous modes, at constant light supply and constant gas-to-culture volume ratio, but at varying CO₂-to-air ratios. Highest biomass production was recorded at CO₂-to-air ratio of 4% with Scenedesmus sp. and at 1% with Nannochloropsis salina. The energetic performance of this IIPBR was quantified in terms of biomass productivity per unit energy input, P/E (g W^?1 day^?1), considering energy input for illumination and for pneumatic mixing and circulation. Under optimal conditions, the IIPBR evaluated in this study achieved P/E of 1.42 g W^?1 day^?1 for Scenedesmus sp. and P/E of 0.34 g W^?1 day^?1 for Nannochloropsis salina. These P/E values are better than those estimated for airlift and bubble column photobioreactor configurations reported in the literature.     

Evaluation of growth and phycobiliprotein composition of cyanobacteria isolates cultivated in different nitrogen sources

Phycobiliproteins, light-harvesting pigments found in cyanobacteria and in some eukaryotic algae, have numerous commercial applications in food, cosmetic, and pharmaceutical industries. Colorant production from cyanobacteria offers advantages over their production from higher plants, as cyanobacteria have fast growth rate and high photosynthetic efficiency and require less space. In this study, three cyanobacteria strains were studied for phycobiliprotein production and the influence of sodium nitrate, potassium nitrate and ammonium chloride on the growth and phycobiliprotein composition of the strains were evaluated. In the batch culture period of 12 days, Phormidium sp. and Pseudoscillatoria sp. were able to utilize all tested nitrogen sources; however, ammonium chloride was the best nitrogen source for both strains to achieve maximum growth rate μ?=?0.284?±?0.03 and μ?=?0.274?±?0.13 day^?1, chlorophyll a 16.2?±? 0.5 and 12.2?±? 0.2 mg L^?1, and phycobiliprotein contents 19.38?±?0.09 and 19.99?±?0.14% of dry weight, whereas, for Arthrospira platensis, the highest growth rate of μ?=?0.304?±?0.0 day^?1, chlorophyll a 19.1?±?0.5 mg L^?1, and phycobiliprotein content of 22.27?±?0.21% of dry weight were achieved with sodium nitrate. The phycocyanin from the lyophilized cyanobacterial biomass was extracted using calcium chloride and food grade purity (A₆₂₀/A₂₈₀ ratio >?0.7) was achieved. Furthermore, phycocyanin was purified using two-step chromatographic method and the analytical grade purity (A₆₂₀/A₂₈₀ ratio >?4) was attained. SDS-PAGE demonstrated the purity and presence of two bands corresponding to α- and β-subunits of the C-phycocyanin. The results showed that Phormidium sp. and Pseudoscillatoria sp. could be good candidates for phycocyanin production.     

Biomass and terpenoids produced by mutant strains of <Emphasis Type="Italic">Arthrospira</Emphasis> under low temperature and light conditions

The filamentous Cyanobacterium Arthrospira is commercially produced and is a functional, high-value, health food. We identified 5 low temperature and low light intensity tolerant strains of Arthrospira sp. (GMPA1, GMPA7, GMPB1, GMPC1, and GMPC3) using ethyl methanesulfonate mutagenesis and low temperature screening. The 5 Arthrospira strains grew rapidly below 14?°C, 43.75 μmol photons m^?2 s^?1 and performed breed conservation at 2.5?°C, 8.75 μmol photons m^?2 s^?1. We used morphological identification and molecular genetic analysis to identify GMPA1, GMPA7, GMPB1 and GMPC1 as Arthrospira platensis, while GMPC3 was identified as Arthrospira maxima. Growth at different culture temperatures was determined at regular intervals using dry biomass. At 16?°C and 43.75 μmol photons m^?2 s^?1, the maximum dry biomass production and the mean dry biomass productivity of GMPA1, GMPB1, and GMPC1 were 2057?±?80 mg l^?1, 68.7?±?2.5 mg l^?1 day^?1, 1839?±?44 mg l^?1, 60.6?±?1.8 mg l^?1 day^?1, and 2113?±?64 mg l^?1, 77.7?±?2.5 mg l^?1 day^?1 respectively. GMPB1 was chosen for additional low temperature tolerance studies and growth temperature preference. In winter, GMPB1 grew well at mean temperatures <10?°C, achieving 3258 mg dry biomass from a starting 68 mg. In summer, GMPB1 grew rapidly at mean temperatures more than 28?°C, achieving 1140 mg l^?1 dry biomass from a starting 240 mg. Phytonutrient analysis of GMPB1 showed high levels of C-phycocyanin and carotenoids. Arthrospira metabolism relates to terpenoids, and the methyl-d-erythritol 4-phosphate pathway is the only terpenoid biosynthetic pathway in Cyanobacteria. The 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) gene from GMPB1 was cloned and phylogenetic analysis showed that GMPB1 is closest to the Cyanobacterium Oscillatoria nigro-viridis PCC711. Low temperature tolerant Arthrospira strains could broaden the areas suitable for cultivation, extend the seasonal cultivation time, and lower production costs.     

Phycoremediation potential,physiological, and biochemical response of <Emphasis Type="Italic">Amphora subtropica</Emphasis> and <Emphasis Type="Italic">Dunaliella</Emphasis> sp. to nickel pollution

Metal pollution can produce many biological effects on aquatic environments. The marine diatom Amphora subtropica and the green alga Dunaliella sp. possess a high metal absorption capacity. Nickel (Ni) removal by living cells of A. subtropica and Dunaliella sp. was tested in cultures exposed to different Ni concentrations (100, 200, 300, and 500 mg L^?1). The amount of Ni removed by the microalgae increased with the time of exposure and the initial Ni concentration in the medium. The metal, which was mainly removed by bioadsorption to Dunaliella sp. cell surfaces (93.63% of total Ni (for 500 mg Ni L^?1) and by bioaccumulation (80.82% of total Ni (for 300 mg Ni L^?1) into Amphora subtropica cells, also inhibited growth. Exposure to Ni drastically reduced the carbohydrate and protein concentrations and increased total lipids from 6.3 to 43.1 pg cell^?1, phenolics 0.092 to 0.257 mg GAE g^?1 (Fw), and carotenoid content, from 0.08 to 0.59 mg g^?1 (Fw), in A. subtropica. In Dunaliella sp., total lipids increased from 26.1 to 65.3 pg cell^?1, phenolics from 0.084 to 0.289 mg GAE g^?1 (Fw), and carotenoid content from 0.41 to 0.97 mg g^?1 (Fw). These compounds had an important role in protecting the algae against ROS generated by Ni. In order to cope with Ni stress shown by the increase of TBARS level, enzymatic (SOD, CAT, and GPx) ROS scavenging mechanisms were induced.     

Characterization of cell structural change,growth, lipid accumulation,and pigment profile of a novel oleaginous microalga, <Emphasis Type="Italic">Vischeria stellata</Emphasis> (Eustigmatophyceae), cultured with different initial nitrate supplies

The appropriate microalgal species and the optimal nitrogen supply in culture medium are vital factors in maximizing biomass and metabolite productivities in microalgae. Vischeria stellata is an edaphic unicellular eustigmatophycean microalga. Cytological and ultrastructural characteristics and the effects of different initial nitrate-nitrogen concentrations on growth, lipid accumulation, fatty acid profile, and pigment composition were investigated in the present study. The cell structures of V. stellata changed with the degree of nutrient utilization and growth phase. The initial nitrate concentration for the optimal growth of V. stellata ranged from 6.0 to 9.0 mM. The maximum total lipid (TLs), neutral lipid (NLs), and total fatty acid (TFAs) contents were 55.9, 51.9, and 44.7 % of dry biomass, respectively. The highest volumetric productivity of TLs, NLs, and TFAs reached 0.28, 0.25, and 0.21 g L^?1 day^?1, respectively. V. stellata had a suitable fatty acid profile for biodiesel production, as well as containing eicosapentaenoic acid (EPA) for nutraceutical applications. In addition, the content β-carotene, increased gradually as culture time was prolonged, resulting in its exclusive production at the end of cultivation. V. stellata is a promising microalgal strain for the production of biofuels and bioproducts.     

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.     

Physiological and Biochemical Mechanisms Preventing Cd Toxicity in the New Hyperaccumulator <Emphasis Type="Italic">Abelmoschus manihot</Emphasis>

Abelmoschus manihot, an ornamental plant, was examined for phytoremediation purposes in accordance with the ability to accumulate cadmium and physiological mechanisms of cadmium tolerance. A net photosynthetic rate (A _N) glasshouse experiment for 60 days was conducted to investigate the influence of different cadmium amounts (0–100 mg kg^?1) on the growth, biomass, photosynthetic performance, reactive oxygen species (ROS) production, antioxidative enzyme activities, Cd uptake and accumulation of A. manihot. Exposure to cadmium enhanced plant growth even at 100 mg kg^?1, without showing symptoms of visible damage. The cadmium concentration of shoots (stems or leaves) and roots was more than the critical value of 100 mg kg^?1 and reached 126.17, 185.26 and 210.24 mg kg^?1, respectively. BCF values of A. manihot plants exceeded the reference value 1.0 for all the Cd treatments, and TF values were greater than 1 at 15–60 mg kg^?1 Cd treatment. The results also showed that cadmium concentrations of 60 mg kg^?1 or less induced a significant enhancement in plant net photosynthetic rate (A _N), stomatal conductance (G _s), transpiration rate (T _r), photosynthetic pigments and F _v/F _m. These parameters were slightly decreased at the higher concentration (100 mg kg^?1). The ROS production (O₂ ^?, H₂O₂) and antioxidative response including SOD, CAT and POD were significantly enhanced by increasing cadmium. These results suggest that A. manihot can be considered as a Cd-hyperaccumulator and the hormetic effects may be taken into consideration in remediation of Cd contamination soil.     

Improved production of carotenoid-free welan gum in a genetic-engineered <Emphasis Type="Italic">Alcaligenes</Emphasis> sp. ATCC31555

Objective

To improve the production of welan gum and obtain a carotenoid-free strain while reducing the fermentation and post-treatment costs.

Results

The vitreoscilla globin (vgb) gene combined with the β-galactosidase (lacZ) promoter was inserted into the phytoene synthase (crtB) gene region of the chromosome in Alcaligenes sp. ATCC31555. When the recombinant strain was grown in a 5 l fermentor, welan gum was produced at 24 ± 0.4 g l^?1 compared to 21 g ± 0.4 g l^?1 in the wild type. Furthermore, the carotenoid-free welan gum produced using Alcaligenes sp. ATCC31555 VHb strain was less expensive with improved properties.

Conclusions

Alcaligenes sp. ATCC31555 VHb strain was a better neutral welan-producing strain with a higher production than the wild-type strain.

     

The growth of <Emphasis Type="Italic">Chlorella sorokiniana</Emphasis> as influenced by CO<Subscript>2</Subscript>, light,and flue gases

In order to achieve recognition as environmentally friendly production, flue gases should be used as a CO₂ source for growing the microalgae Chlorella sorokiniana when used for hydrogen production. Flue gases from a waste incinerator and from a silicomanganese smelter were used. Before testing the flue gases, the algae were grown in a laboratory at 0.04, 1.3, 5.9, and 11.0 % (v/v) pure CO₂ gas mixed with fresh air. After 5 days of growth, the dry biomass per liter algal culture reached its maximum at 6.1 % CO₂. A second experiment was conducted in the laboratory at 6.2 % CO₂ at photon flux densities (PFD) of 100, 230, and 320 μmol photons m^?2 s^?1. After 4 days of growth, increasing the PFD increased the biomass production by 67 and 108 % at the two highest PFD levels, as compared with the lowest PFD. A bioreactor system containing nine daylight-exposed tubes and nine artificial light-exposed tubes was installed on the roof of the waste incinerator. The effect of undiluted flue gas (10.7 % CO₂, 35.8 ppm NO _x , and 38.6 ppm SO₂), flue gas diluted with fresh air to give 4.2 % CO₂ concentration, and 5.0 % pure CO₂ gas was studied in daylight (21.4?±?9.6 mol photons m^?2 day^?1 PAR, day length 12.0 h) and at 135 μmol photons m^?2 s^?1 artificial light given 24 h day^?1 (11.7?±?0.0 mol photons m^?2 day^?1 PAR). After 4 days’ growth, the biomass production was the same in the two flue gas concentrations and the 5 % pure CO₂ gas control. The biomass production was also the same in daylight and artificial light, which meant that, in artificial light, the light use efficiency was about twice that of daylight. The starch concentration of the algae was unaffected by the light level and CO₂ concentration in the laboratory experiments (2.5–4.0 % of the dry weight). The flue gas concentration had no effect on starch concentration, while the starch concentration increased from about 1.5 % to about 6.0 % when the light source changed from artificial light to daylight. The flue gas from the silicomanganese smelter was characterized by a high CO₂ concentration (about 17 % v/v), low oxygen concentration (about 4 %), about 100 ppm NO _x , and 1 ppm SO₂. The biomass production using flue gas significantly increased as compared with about 5 % pure CO₂ gas, which was similar to the biomass produced at a CO₂ concentration of 10–20 % mixed with N₂. Thus, the enhanced biomass production seemed to be related to the low oxygen concentration rather than to the very high CO₂ concentration.     

The effect of temperature on growth and lipid and fatty acid composition on marine microalgae used for biodiesel production

Cultivation temperature is one of the major factors affecting the growth and lipid accumulation of microalgae. In this study, the effects of temperature on the growth, lipid content, fatty acid composition and biodiesel properties of the marine microalgae Chaetoceros sp. FIKU035, Tetraselmis suecica FIKU032 and Nannochloropsis sp. FIKU036 were investigated. These species were cultured at different temperatures (25, 30, 35 and 40 °C). The results showed that the specific growth rate, biomass and lipid content of all microalgae decreased with increasing temperature. With regards to fatty acids, the presence of saturated fatty acids (SFAs) in T. suecica FIKU032 and Nannochloropsis sp. FIKU036 decreased with increasing temperature, in contrast with polyunsaturated fatty acids (PUFAs). Moreover, Chaetoceros sp. FIKU035 was the only species that could grow at 40 °C. The highest lipid productivity was observed in Chaetoceros sp. FIKU035 when cultivated at 25 °C (66.73 ± 1.34 mg L^?1 day^?1) and 30 °C (61.35 ± 2.89 mg L^?1 day^?1). Moreover, the biodiesel properties (cetane number, cold filter plugging point, kinematic viscosity and density) of the lipids obtained from this species were in accordance with biodiesel standards. This study indicated that Chaetoceros sp. FIKU035 can be considered as a suitable species for biodiesel production in outdoor cultivation.     

<Emphasis Type="SmallCaps">d</Emphasis>-Xylose fermentation,xylitol production and xylanase activities by seven new species of <Emphasis Type="Italic">Sugiyamaella</Emphasis>

Sixteen yeast isolates identified as belonging to the genus Sugiyamaella were studied in relation to D-xylose fermentation, xylitol production, and xylanase activities. The yeasts were recovered from rotting wood and sugarcane bagasse samples in different Brazilian regions. Sequence analyses of the internal transcribed spacer (ITS) region and the D1/D2 domains of large subunit rRNA gene showed that these isolates belong to seven new species. The species are described here as Sugiyamaella ayubii f.a., sp. nov. (UFMG-CM-Y607^T = CBS 14108^T), Sugiyamaella bahiana f.a., sp. nov. (UFMG-CM-Y304^T = CBS 13474^T), Sugiyamaella bonitensis f.a., sp. nov. (UFMG-CM-Y608^T = CBS 14270^T), Sugiyamaella carassensis f.a., sp. nov. (UFMG-CM-Y606^T = CBS 14107^T), Sugiyamaella ligni f.a., sp. nov. (UFMG-CM-Y295^T = CBS 13482^T), Sugiyamaella valenteae f.a., sp. nov. (UFMG-CM-Y609^T = CBS 14109^T) and Sugiyamaella xylolytica f.a., sp. nov. (UFMG-CM-Y348^T = CBS 13493^T). Strains of the described species S. boreocaroliniensis, S. lignohabitans, S. novakii and S. xylanicola, isolated from rotting wood of Brazilian ecosystems, were also compared for traits relevant to xylose metabolism. S. valenteae sp. nov., S. xylolytica sp. nov., S. bahiana sp. nov., S. bonitensis sp. nov., S. boreocarolinensis, S. lignohabitans and S. xylanicola were able to ferment d-xylose to ethanol. Xylitol production was observed for all Sugiyamaella species studied, except for S. ayubii sp. nov. All species studied showed xylanolytic activity, with S. xylanicola, S. lignohabitans and S. valenteae sp. nov. having the highest values. Our results suggest these Sugiyamaella species have good potential for biotechnological applications.     

Changes in growth and photosynthesis of mixotrophic <Emphasis Type="Italic">Ochromonas</Emphasis> sp. in response to different concentrations of glucose

Ochromonas sp. (Chrysophyceae) occupies different trophic levels depending on its prevalent nutritional strategy in the field. Ochromonas sp. can utilize inorganic and organic carbon to grow photoautotrophically, mixotrophically, and heterotrophically. To examine how exogenous organic carbon influences population growth and photosynthesis in Ochromonas sp., we cultured Ochromonas sp. in media with different glucose concentrations (0, 50, 100, and 150 mg L^?1) and two treatment modes (single addition of glucose at the beginning and fed-batch mode with repeated addition every 12 h) for 15 days. Results showed that (1) compared to photoautotrophic growth, adding glucose significantly enhanced the growth rate and carrying capacity of Ochromonas sp. in a concentration-dependent manner; (2) photosynthesis also increased with glucose addition during the exponential growth stage; (3) repeated glucose addition caused higher biomass and photosynthetic performances than single addition, and (4) with depletion of glucose and the population reaching a stationary stage, photosynthetic parameters decreased slightly but remained at a relatively stable level, which was similar to that of photoautotrophic growth. We conclude that in comparison with pure photoautotrophy, mixotrophic growth can result in higher population growth rate and final biomass in a shorter time, especially in fed-batch cultures, indicating that this would be a highly efficient way for mass cultivation of the flagellate Ochromonas.     

Single point mutations enhance activity of <Emphasis Type="Italic">cis</Emphasis>-epoxysuccinate hydrolase

Objectives

To enhance activity of cis-epoxysuccinate hydrolase from Klebsiella sp. BK-58 for converting cis-epoxysuccinate to tartrate.

Results

By semi-saturation mutagenesis, all the mutants of the six important conserved residues almost completely lost activity. Then random mutation by error-prone PCR and high throughput screening were further performed to screen higher activity enzyme. We obtained a positive mutant F10D after screening 6000 mutations. Saturation mutagenesis on residues Phe10 showed that most of mutants exhibited higher activity than the wild-type, and the highest mutant was F10Q with activity of 812 U mg^?1 (k _cat /K _m , 9.8 ± 0.1 mM^?1 s^?1), which was 230 % higher than that of wild-type enzyme 355 U mg^?1 (k _cat /K _m , 5.3 ± 0.1 mM^?1 s^?1). However, the thermostability of the mutant F10Q slightly decreased.

Conclusions

The catalytic activity of a cis-epoxysuccinate hydrolase was efficient improved by a single mutation F10Q and Phe10 might play an important role in the catalysis.

     

Magnetophoretic harvesting of freshwater microalgae using polypyrrole/Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> nanocomposite and its reusability

A magnetophoretic harvesting agent, a polypyrrole/Fe₃O₄ magnetic nanocomposite, is proposed as a cost and energy efficient alternative to recover biomass of the microalgae Botryococcus braunii, Chlorella protothecoides, and Chlorella vulgaris from their culture media. The maximal recovery efficiency reached almost 99 % for B. braunii, 92.4 % for C. protothecoides, and 90.8 % for C. vulgaris. The maximum adsorption capacity (Q ₀) of the magnetic nanocomposite for B. braunii (63.49 mg dry biomass mg^?1 PPy/Fe₃O₄) was higher than that for C. protothecoides (43.91 mg dry biomass mg^?1 PPy/Fe₃O₄) and C. vulgaris (39.98 mg dry biomass mg^?1 PPy/Fe₃O₄). The highest harvesting efficiency for all the studied microalgae were at pH 10.0, and measurement of zeta-potential confirmed that the flocculation was induced by charge neutralization. This study showed that polypyrrole/Fe₃O₄ can be a promising flocculant due to its high efficacy, low dose requirements, short settling time, its integrity with cells, and with great potential for saving energy because of its recyclability.