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.     

Impact of elevated CO<Subscript>2</Subscript> in <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> rooted stem cuttings inoculated with <Emphasis Type="Italic">Frankia</Emphasis>

Impact of different levels of elevated CO ₂ on the activity of Frankia (Nitrogen-fixing actinomycete) in Casuarina equisetifolia rooted stem cuttings has been studied to understand the relationship between C. equisetifolia, Frankia and CO₂. The stem cuttings of C. equietifolia were collected and treated with 2000 ppm of Indole Butyric Acid (IBA) for rooting. Thus vegetative propagated rooted stem cuttings of C. equisetifolia were inoculated with Frankia and placed in the Open top chambers (OTC) with elevated CO₂ facilities. These planting stocks were maintained in the OTC for 12 months under different levels of elevated CO₂ (ambient control, 600 ppm, 900 ppm). After 12 months, the nodule numbers, bio mass, growth, and photosynthesis of C. equisetifolia rooted stem cuttings inoculated with Frankia were improved under 600 ppm of CO₂. The rooted stem cuttings of C. equisetifolia inoculated with Frankia showed a higher number of nodules under 900 ppm of CO₂ and cuttings without Frankia inoculation exhibited poor growth. Tissue Nitrogen (N) content was also higher under 900 ppm of CO₂ than ambient control and 600 ppm levels. The photosynthetic rate was higher (17.8 μ mol CO₂ m^?2 s^?1) in 900 ppm of CO₂ than in 600 ppm (13.2 μ mol CO₂ m^?2 s^?1) and ambient control (8.3 μ mol CO₂ m^?2 s^?1). This study showed that Frankia can improve growth, N fixation and photosynthesis of C. equietifolia rooted stem cuttings under extreme elevated CO₂ level conditions (900 ppm).     

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.     

<Emphasis Type="Italic">Tetraselmis suecica</Emphasis> culture for CO<Subscript>2</Subscript> bioremediation of untreated flue gas from a coal-fired power station

The accumulation of atmospheric CO₂, primarily due to combustion of fossil fuels, has been implicated in potential global climate change. The high rate of CO₂ bioremediation by microalgae has emerged as a favourable method for reducing coal-fired power plant emissions. However, coal-fired power station flue gas contains other chemicals such as SO_x which can inhibit microalgal growth. In the current study, the effect of untreated flue gas as a source of inorganic carbon on the growth of Tetraselmis in a 1000 L industrial-scale split-cylinder internal-loop airlift photobioreactor was examined. The culture medium was recycled after each harvest. Tetraselmis suecica grew very well in this airlift photobioreactor during the 7-month experiment using recycled medium from an electroflocculation harvesting unit. Increased medium SO₄ ^2? concentration as high as 870 mg SO₄ ^2??L^?1 due to flue gas addition and media recycling had no negative effect on the overall growth and productivity of this alga. The potential organic biomass productivity and carbon sequestration using an industrial-scale airlift PBR at International Power Hazelwood, Gippsland, Victoria, Australia, are 178.9?±?30 mg L^?1 day^?1 and 89.15?±?20 mg?‘C’?L^?1 day^?1, respectively. This study clearly indicates the potential of growing Tetraselmis on untreated flue gas and using recycled medium for the purpose of biofuel and CO₂ bioremediation.     

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.     

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.     

Biological CO<Subscript>2</Subscript> fixation using C<Emphasis Type="Italic">hlorella vulgaris</Emphasis> and its thermal characteristics through thermogravimetric analysis

The present research is focused on cultivation of microalgae strain Chlorella vulgaris for bio-fixation of CO₂ coupled with biomass production. In this regard, a single semi-batch vertical tubular photobioreactor and four similar photobioreactors in series have been employed. The concentration of CO₂ in the feed stream was varied from 2 to 12 % (v/v) by adjusting CO₂ to air ratio. The amount of CO₂ capture and algae growth were monitored by measuring decrease of CO₂ concentration in the gas phase, microalgal cell density, and algal biomass production rate. The results show that 4 % CO₂ gives maximum amount of biomass (0.9 g L^?1) and productivity (0.118 g L^?1 day^?1) of C. vulgaris in a single reactor. In series reactors, average productivity per reactor found to be 0.078 g L^?1 day^?1. The maximum CO₂ uptake for single reactor also found with 4 % CO_2, and it is around 0.2 g L^?1 day^?1. In series reactors, average CO₂ uptake is 0.13 g L^?1 day^?1 per reactor. TOC analysis shows that the carbon content of the produced biomass is around 40.67 % of total weight. The thermochemical characteristics of the cultivated C. vulgaris samples were analyzed in the presence of air. All samples burn above 200 °C and the combustion rate become faster at around 600 °C. Almost 98 wt% of the produced biomass is combustible in this range.     

Carbon Dioxide and Methane Fluxes From Tree Stems,Coarse Woody Debris,and Soils in an Upland Temperate Forest

Forest soils and canopies are major components of ecosystem CO₂ and CH₄ fluxes. In contrast, less is known about coarse woody debris and living tree stems, both of which function as active surfaces for CO₂ and CH₄ fluxes. We measured CO₂ and CH₄ fluxes from soils, coarse woody debris, and tree stems over the growing season in an upland temperate forest. Soils were CO₂ sources (4.58 ± 2.46 µmol m^?2 s^?1, mean ± 1 SD) and net sinks of CH₄ (?2.17 ± 1.60 nmol m^?2 s^?1). Coarse woody debris was a CO₂ source (4.23 ± 3.42 µmol m^?2 s^?1) and net CH₄ sink, but with large uncertainty (?0.27 ± 1.04 nmol m^?2 s^?1) and with substantial differences depending on wood decay status. Stems were CO₂ sources (1.93 ± 1.63 µmol m^?2 s^?1), but also net CH₄ sources (up to 0.98 nmol m^?2 s^?1), with a mean of 0.11 ± 0.21 nmol m^?2 s^?1 and significant differences depending on tree species. Stems of N. sylvatica, F. grandifolia, and L. tulipifera consistently emitted CH₄, whereas stems of A. rubrum, B. lenta, and Q. spp. were intermittent sources. Coarse woody debris and stems accounted for 35% of total measured CO₂ fluxes, whereas CH₄ emissions from living stems offset net soil and CWD CH₄ uptake by 3.5%. Our results demonstrate the importance of CH₄ emissions from living stems in upland forests and the need to consider multiple forest components to understand and interpret ecosystem CO₂ and CH₄ dynamics.     

Short-term effects of crop rotation,residue management,and soil water on carbon mineralization in a tropical cropping system

The purpose of this study was to investigate the short-term effects of maize (Zea mays)-fallow rotation, residue management, and soil water on carbon mineralization in a tropical cropping system in Ghana. After 15 months of the trial, maize–legume rotation treatments had significantly (P?C ₀ (μg CO₂–C g^?1) than maize–elephant grass (Pennisetum purpureum) rotations. The C ₀ for maize–grass rotation treatments was significantly related to the biomass input (r?=?0.95; P?=?0.05), but that for the maize–legume rotation was not. The soil carbon mineralization rate constant, k (per day), was also significantly related to the rotation treatments (P?k values for maize–grass and maize–legume rotation treatments were 0.025 and 0.036 day^?1 respectively. The initial carbon mineralization rate, m ₀ (μg CO₂–C g^?1 day ^?1), was significantly (P?θ. The m ₀ ranged from 3.88 to 18.67 and from 2.30 to 15.35 μg CO₂–C g^?1 day^?1 for maize–legume and maize–grass rotation treatments, respectively, when the soil water varied from 28% to 95% field capacity (FC). A simple soil water content (θ)-based factor, f _w, formulated as: \(f_{\text{w}} = \left[ {\frac{{\theta - \theta _{\text{d}} }}{{\theta _{{\text{FC}}} - \theta _{\text{d}} }}} \right]\), where θ _d and θ _FC were the air-dry and field capacity soil water content, respectively, adequately described the variation of the m ₀ with respect to soil water (R ²?=?0.91; RMSE?=?1.6). Such a simple relationship could be useful for SOC modeling under variable soil water conditions.     

General medium for the autotrophic cultivation of acetogens

Syngas fermentation, a microbial process in which synthesis gas serves as a substrate for acetogens, has attracted increasing interest in the last few years. For the purposeful selection of acetogens for various applications, it would be useful to characterize and compare the process performances of as many autotrophic strains as possible under identical process conditions. Unfortunately, all the media compositions so far recommended for syngas fermentation differ considerably with respect to each individual strain. Therefore, a general medium for syngas fermentation was designed. The suitability of this new general-acetogen medium (GA-medium) was proven based on the autotrophic batch cultivation of Acetobacterium fimetarium, Acetobacterium wieringae, Blautia hydrogenotrophica, Clostridium magnum, Eubacterium aggregans, Sporomusa acidovorans, Sporomusa ovata and Terrisporobacter mayombei in anaerobic flasks with an initial gas phase of H₂:CO₂ (66:34) (P = 200 kPa). A comparison of the autotrophic batch processes with this medium revealed T. mayombei as the bacterium with the highest maximum growth rate of 5.77 day^?1 which was more than 10 times higher than the lowest identified maximum growth rate of A. fimetarium. The maximum growth rates of A. wieringae, C. magnum and S. acidovorans were all in the same order of magnitude around 1.7 day^?1. The newly designed GA-medium offers the possibility to compare autotrophic process performances of different acetogens under similar conditions absent the effects of various media compositions.     

Population Growth Parameters of Rose Aphid, <Emphasis Type="Italic">Macrosiphum rosae</Emphasis> (Hemiptera: Aphididae) on Different Rose Cultivars

The rose aphid, Macrosiphum rosae (L.), is one of the most important pests on rose plants (Rosa spp.) with a worldwide distribution. As resistance indices, the development, survivorship, and reproduction of this aphid were evaluated on 10 rose cultivars, including Bella Vita, Cool Water, Dolce Vita, Maroussia, Orange Juice, Pinkpromise, Roulette, Tea, Valentine, and Persian Yellow in laboratory at 25?±?1°C, 65?±?5% relative humidity, and photoperiod of 16:8 (L/D)?h. Rose aphid successfully survived on all 10 rose cultivars, although mortality rate was higher on Tea and Bella Vita. The number of offspring per female differed significantly among the tested rose cultivars, and ranged from 9.2 on Tea to 38.7 nymphs on Orange Juice. Population growth parameters were significantly affected by rose cultivars. The longest mean generation time (T) was observed on Bella Vita (14.8 days) and Tea (14.7 days) and the shortest on Orange Juice (10.0 days). The net reproductive rate (R ₀ ) ranged from 6.9 on Tea to 33.2 nymphs on Orange Juice cultivar. Correspondingly, the highest value of intrinsic rate of increase (r _m ) was observed on Orange Juice (0.348 day^?1) and lower values on Tea (0.131 day^?1) followed by Bella Vita (0.154 day^?1). Cluster analysis of all the measured parameters of rose aphid on different rose cultivars revealed that Tea and Bella Vita were relatively resistant to M. rosae. These findings could be useful in developing an integrated pest management (IPM) program for this aphid in urbanized areas and commercial rose potting.     

Application of <Emphasis Type="Italic">Chlorella sorokiniana</Emphasis> (Chlorophyceae) as supplement and/or an alternative medium for the in vitro cultivation of <Emphasis Type="Italic">Schomburgkia crispa</Emphasis> (Orchidaceae)

Schomburgkia crispa Lindley (Orchidaceae) is an epiphytic species found in gallery forests and dry vegetation in the Brazilian Cerrado. It is typically unable to germinate or exhibits low germination because of dependency on mycorrhizal associations. In vitro cultivation techniques have helped circumvent difficulties involved in propagation from seeds. Alternative media and organic biostimulant substances that reduce costs and promote satisfactory in vitro growth are constantly sought. This study evaluated in vitro multiplication and rooting of S. crispa in a modified culture medium containing extract of the microalga Chlorella sorokiniana. We analyzed supplementation of WPM (Woody Plant Medium) with microalgae suspended in NPK medium, or as the supernatant resulting from the centrifugation of a culture in NPK medium. The extracts were added to WPM instead of distilled water. The compounds 6-benzylaminopurine (BAP) and indolebutyric acid (IBA) were used as reference in the in vitro multiplication and rooting of S. crispa, respectively. Both growth regulators were tested at 0, 2.5, and 5.0 mg L^?1. During in vitro multiplication of S. crispa, WPM supplemented with 5.0 mg L^?1 BAP favored the formation of more sprouts, whereas WPM containing 2.5 mg L^?1 IBA supplemented with microalgae extract stimulated in vitro rooting. Schomburgkia crispa explants cultivated in medium supplemented with microalgae suspension or the supernatant of C. sorokiniana showed growth similar to explants cultivated in WPM alone. Therefore, it is possible to use the microalga C. sorokiniana as a supplement and/or alternative to WPM for the in vitro cultivation of S. crispa.     

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.     

Characterization of alcohol dehydrogenase from <Emphasis Type="Italic">Kangiella koreensis</Emphasis> and its application to production of all-<Emphasis Type="Italic">trans</Emphasis>-retinol

A recombinant alcohol dehydrogenase (ADH) from Kangiella koreensis was purified as a 40 kDa dimer with a specific activity of 21.3 nmol min^?1 mg^?1, a K _m of 1.8 μM, and a k _cat of 1.7 min^?1 for all-trans-retinal using NADH as cofactor. The enzyme showed activity for all-trans-retinol using NAD ⁺ as a cofactor. The reaction conditions for all-trans-retinol production were optimal at pH 6.5 and 60 °C, 2 g enzyme l^?1, and 2,200 mg all-trans-retinal l^?1 in the presence of 5 % (v/v) methanol, 1 % (w/v) hydroquinone, and 10 mM NADH. Under optimized conditions, the ADH produced 600 mg all-trans-retinol l^?1 after 3 h, with a conversion yield of 27.3 % (w/w) and a productivity of 200 mg l^?1 h^?1. This is the first report of the characterization of a bacterial ADH for all-trans-retinal and the biotechnological production of all-trans-retinol using ADH.     

Utilization of gaseous emissions from a methanol plant for cultivation of <Emphasis Type="Italic">Acutodesmus obliquus</Emphasis>

This study aimed to culture the green alga Acutodesmus obliquus utilizing the gaseous emissions containing a high concentration of CO₂ (99.13 %) from a methanol plant and study the tolerance of microalgae. The effect of CO₂ concentration, aeration rate, inoculum concentration, intermittent sparging, and nitrogen sources on the growth of A. obliquus was examined. Acutodesmus obliquus also was cultivated in a 500-L pilot outdoor tubular photobioreactor (OTP) to advance the laboratory scale system to outdoor scale-up applications. The results showed that A. obliquus could tolerate high CO₂ concentrations of 50 %, and a maximum biomass of 0.935 g L^?1 (dry weight) was achieved at 20 % CO₂. An aeration rate of 500 mL min^?1, inoculum concentration (optical density at 680 nm [OD₆₈₀]?=?0.3), and intermittent sparging of 10 min per 2 h enhanced growth to the optimum and influenced culture pH and photosynthesis. Urea as a nitrogen source was shown to be more beneficial to cell growth. A urea concentration of 0.3 g L^?1 and an N/P ratio of 15 led to maximum biomass accumulation thus enhancing the gaseous emission utilization efficiency. In conclusion, this work demonstrated that gaseous emissions containing high concentration of CO₂ from a methanol plant could be directly introduced into A. obliquus cultures and that A. obliquus was suitable well for large-scale outdoor cultivation in a tubular photobiorecator.     

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.

     

Photosynthetic CO<Subscript>2</Subscript> uptake in seedlings of two tropical tree species exposed to oscillating elevated concentrations of CO<Subscript>2</Subscript>

Do short-term fluctuations in CO₂ concentrations at elevated CO₂ levels affect net CO₂ uptake rates of plants? When exposed to 600 μl CO₂ l^?1, net CO₂ uptake rates in shoots or leaves of seedlings of two tropical C₃ tree species, teak (Tectona grandis L. f.) and barrigon [Pseudobombax septenatum (Jacq.) Dug.], increased by 28 and 52% respectively. In the presence of oscillations with half-cycles of 20 s, amplitude of ca. 170 μl CO₂ l^?1 and mean of 600 μl CO₂ l^?1, the stimulation in net CO₂ uptake by the two species was reduced to 19 and 36%, respectively, i.e. the CO₂ stimulation in photosynthesis associated with a change in exposure from 370 to 600 μl CO₂ l^?1 was reduced by a third in both species. Similar reductions in CO₂-stimulated net CO₂ uptake were observed in T. grandis exposed to 40-s oscillations. Rates of CO₂ efflux in the dark by whole shoots of T. grandis decreased by 4.8% upon exposure of plants grown at 370 μl CO₂ l^?1 to 600 μl CO₂ l^?1. The potential implications of the observations on CO₂ oscillations and dark respiration are discussed in the context of free-air CO₂ enrichment (FACE) systems in which short-term fluctuations of CO₂ concentration are a common feature.     

Optimization of continuous-flow solid-phase denitrification via coupling carriers in enhancing simultaneous removal of nitrogen and organics for agricultural runoff purification

Coupling of biodegradable corncob and plastic carrier was optimized in continuous-flow solid-phase denitrification systems for enhancing simultaneously removal of nitrogen and organics in agricultural runoff. In compared with preposition of plastic carriers and mixed distribution method, it was demonstrated that the preposition of corncobs simultaneously enhanced nitrate (6.64 ± 1.35 mg L^?1 day ^?1) and organics removal (6.33 ± 1.44 mg L^?1 day^?1) at a hydraulic retention time (HRT) of 6 h. The operation performance could be further enhanced with extension of HRT to 12 h. The dominant genera found in corncob were denitrifiers for nitrate reduction (Bosea, Simplicispira, Desulfovibrio, Klebsiella, etc.) and fermentative bacteria (Pleomorphomonas, Actinotalea, Opitutus, Cellulomonas, Bacteroides, etc.) responsible for corncob degrading to simple organics for other denitrifiers. However, much lower and different denitrifiers abundances (Bradyrhizobium, Acinetobacter, Bacillus, etc.) exhibited on plastic filler than those of corncob. It well explained that the biofilm on plastic carrier was mainly related with organics removal while the biofilm on corncobs inclined to effectively remove nitrate, and simultaneous removal of nitrogen and organics could be achieved in coupling carriers system with preposition of biodegradable corncob.     

Canola quality affects second (<Emphasis Type="Italic">Brevicoryne brassicae</Emphasis>) and third (<Emphasis Type="Italic">Diaeretiella rapae</Emphasis>) trophic levels

Biological control agents can be used as a complementary control measure that can be combined with resistant host plants to control pests. In this study, the effects of different canola cultivars (Karaj-1, Karaj-2, Karaj-3, Licord, Okapi, Opera, RGS₀₀₃, Sarigol, Talaye and Zarfam) on the performance and life table parameters of the cabbage aphid, Brevicoryne brassicae, and its parasitoid, Diaeretiella rapae, were determined under laboratory conditions. Total fecundity of the cabbage aphid differed with cultivar, with the highest value (59.41 nymphs per female) of this parameter observed on Opera and the lowest (1.67) observed on RGS₀₀₃. The highest and lowest intrinsic rates of increase (r) of the cabbage aphid were observed on Opera (0.331 day^?1) and RGS₀₀₃ (? 0.242 day^?1) cultivars, respectively, suggesting these to be the most susceptible and most resistant cultivars to this pest. However, because the aphid did not settle and feed well on RGS₀₀₃, it was not possible to determine demographic parameters for its parasitoid. Consequently, the Okapi cultivar, which was the most resistant cultivar to the cabbage aphid after RGS₀₀₃, was used in this study to assess the parasitoid wasp. The parasitoid’s intrinsic rate of increase (r) varied from 0.426 day^?1 on the susceptible cultivar (Opera) to 0.341 day^?1 on the resistant canola cultivar Okapi. Aphid performance decreased 93% on the resistant canola cultivar, while parasitoid performance decreased only 20% on the resistant cultivar compared to more susceptible cultivar.