Contribution of Hydrogenase 2 to Stationary Phase H2 Production by Escherichia coli During Fermentation of Glycerol

Escherichia coli has four hydrogenases (Hyd), three genes of which are encoded by the hya, hyb, and hyc operons. The proton-reducing and hydrogen-oxidizing activities of Hyd-2 (hyb) were analyzed in whole cells grown to stationary phase and cell extracts, respectively, during glycerol fermentation using novel double mutants. H₂ production rate at pH 7.5 was decreased by ~3.5- and ~7-fold in hya and hyc (HDK 103) or hyb and hyc (HDK 203) operon double mutants, respectively, compared with the wild type. At pH 6.5, H₂ production decreased by ~2- and ~5-fold in HDK103 and HDK203, respectively, compared with the wild type. At pH 5.5, H₂ production was reduced by ~4.5-fold in the mutants compared with the wild type. The total hydrogen-oxidizing activity was shown to depend on the pH of the growth medium in agreement with previous findings and was significantly reduced in the HDK103 or HDK203 mutants. At pH 7.5, Hyd-2 activity was 0.26 U (mg protein)^?1 and Hyd-1 activity was 0.1 U (mg protein)^?1. As the pH of the growth medium decreased to 6.5, Hyd-2 activity was 0.16 U (mg protein)^?1, and Hyd-1 was absent. Surprisingly, at pH 5.5, there was an increase in Hyd-2 activity (0.33 U mg protein)^?1 but not in that of Hyd-1. These findings show a major contribution of Hyd-2 to H₂ production during glycerol fermentation that resulted from altered metabolism which surprisingly influenced proton reduction.     

Salicylate Degradation by the Fungal Plant Pathogen Sclerotinia sclerotiorum

The fungal plant pathogen Sclerotinia sclerotiorum was studied to determine its ability to degrade salicylate, an important defense-signaling molecule in plants. S. sclerotiorum D-E7 was grown at 25 °C in an undefined medium (50 ml) containing minerals, 0.1 % soytone, 50 mM MES buffer (pH 6.5), 25 mM glucose, and 1 mM salicylate. Glucose, oxalate, and salicylate concentrations were monitored by HPLC. S. sclerotiorum D-E7 was found to be active in salicylate degradation. However, salicylate alone was not growth supportive and, at higher levels (10 mM), inhibited glucose-dependent growth. Biomass formation (130 mg [dry wt] of mycelium per 50 ml of undefined medium), oxalate concentrations (~10 mM), and culture acidification (final culture pH approximated 5) were essentially the same in cultures grown with or without salicylate (1 mM). Time-course analyses revealed that salicylate degradation and glucose consumption were complete after 7 days of incubation and was concomitant with growth. Trace amounts of catechol, a known intermediate of salicylate metabolism, were detected during salicylate degradation. Overall, these results indicated that S. sclerotiorum has the ability to degrade salicylate and that the presence of low levels of salicylate did not affect growth or oxalate production by S. sclerotiorum.     

Sorption of lead and copper from an aqueous phase system by marine-derived Aspergillus species

A total of 47 cultures of Aspergillus representing 13 species were screened for their ability to tolerate 7.5 mM Pb²⁺ and 2 mM Cu²⁺, all of which were positive, with growth of 31 of the cultures being enhanced by low concentrations of lead. The isolates of Aspergillus versicolor, A. niger and A. flavus were tolerant to concentrations as high as 10 - 12.5 mM Pb²⁺ and 3 – 4 mM Cu²⁺. Selected cultures displayed a good sorption capacity of 32 - 41 mg Pb²⁺ and 3.5 - 6.5 mg Cu²⁺ g^-1 dry weight of mycelia, which was improved by alkali pretreatment of the biomass and negatively affected by mild dry heat treatment. The sequestration of the metal occurred mainly by sorption to the cell-surface with very little intracellular uptake. FTIR analysis indicated the involvement of hydroxyl, amino, and carbonyl groups in Pb²⁺ and Cu²⁺ biosorption by fungal biomass of the different species of Aspergillus.     

Effects of saline-alkaline stress on benzo[a]pyrene biotransformation and ligninolytic enzyme expression by <Emphasis Type="Italic">Bjerkandera adusta</Emphasis> SM46

Benzo[a]pyrene (BaP) accumulates in marine organisms and contaminated coastal areas. The biotreatment of waste water using saline-alkaline-tolerant white rot fungi (WRF) represents a promising method for removing BaP under saline-alkaline conditions based on WRF’s ability to produce ligninolytic enzymes. In a pre-screening for degradation of polycyclic aromatic hydrocarbons of 82 fungal strains using Remazol brilliant blue R, Bjerkandera adusta SM46 exhibited the highest tolerance to saline-alkaline stress. Moreover, a B. adusta culture grown in BaP-containing liquid medium exhibited resistance to salinities up to 20 g l^?1. These conditions did not inhibit fungal growth or the expression of manganese peroxidase (MnP) or lignin peroxidase (LiP). The degradation rate also became higher as salinity increased to 20 g l^?1. Fungal growth and enzyme expression were inhibited at a salinity of 35 g l^?1. These inhibitory effects directly decreased the degradation rate (>24 %). The presence of MnSO₄ as an inducer improved the degradation rate and enzyme expression. MnP and LiP activity also increased by seven- and fivefold, respectively. SM46 degraded BaP (38–89 % over 30 days) in an acidic environment (pH 4.5) and under saline-alkaline stress conditions (pH 8.2). Investigating the metabolites produced revealed BaP-1,6-dione as the main product, indicating the important role of ligninolytic enzymes in initializing BaP cleavage. The other metabolites detected, naphthalene acetic acid, hydroxybenzoic acid, benzoic acid, and catechol, may have been ring fission products. The wide range of activities observed suggests that B. adusta SM46 is a potential agent for biodegrading BaP under saline conditions.     

Purification and characterization of a chitinase from Serratia proteamaculans

A chitinase gene from Serratia proteamaculans 18A1 was cloned, sequenced, and expressed in Escherichia coli M15. Recombinant enzyme (ChiA) was purified by Ni-NTA affinity column chromatography. The ChiA gene contains an open reading frame (ORF), encoding an endochitinase with a deduced molecular weight 60 kDa and predicted isoelectric point of 6.35. Comparison of ChiA with other chitinases revealed a modular structure containing an N-terminal PKD-domain, a family 18 catalytic domain and a C-terminal putative chitin-binding domain. Turn over rate (K _cat) of the enzyme was determined using colloidal chitin (49.71 ± 1.15 S^?1) and crystalline β-chitin (17.20 ± 0.83 S^?1) as substrates. The purified enzyme was active over a broad range of pH (pH 4.5–9.0) and temperature (4–70°C) with a peak activity at pH 5.5 and 55°C. However, enzyme activity was found to be stable up to 45°C for longer incubation periods. Purified enzyme was shown to inhibit fungal spore germination and hyphal growth of pathogenic fungi Fusarium oxysporum and Aspergillus niger.     

Effect of Nutrition and Environmental Factors on the Endoparasitic Fungus Esteya vermicola, a Biocontrol Agent Against Pine Wilt Disease

The nematophagous fungus Esteya vermicola has tremendous potential for biological control. This species exhibits strong infectious activity against pinewood nematodes, whereas the study on the effect of nutrition and environmental factors is still of paucity. Carbon (C), nitrogen (N), pH value, temperature, and water activity have great impact on the fungal growth, sporulation, and germination. In nutrition study, the greatest number of conidia (2.36 × 10⁹ per colony) was obtained at the C:N ratio of 100:1 with a carbon concentration 32 g l^?1. In addition, the germination rate and radial growth of E. vermicola were used to evaluate the effects of environmental conditions and they were optimized as following: pH 5.5, 26 °C and water activity of 0.98. Our results also confirmed that variation of environmental factors has a detrimental influence on the efficacy of active conidia and growth of fungus. Moreover, under above optimal condition, the biocontrol efficacy was significantly improved in regard to the increase of adhesive and mortality rate, which highlight the study on the application of E. vermicola as pine wilt disease biocontrol agent.     

Kinetics of growth and caffeine demethylase production of Pseudomonas sp. in bioreactor

The effect of various initial caffeine concentrations on growth and caffeine demethylase production by Pseudomonas sp. was studied in bioreactor. At initial concentration of 6.5 g l^?1 caffeine, Pseudomonas sp. showed a maximum specific growth rate of 0.2 h^?1, maximum degradation rate of 1.1 g h^?1, and caffeine demethylase activity of 18,762 U g CDW^?1 (CDW: cell dry weight). Caffeine degradation rate was 25 times higher in bioreactor than in shake flask. For the first time, we show highest degradation of 75 g caffeine (initial concentration 20 g l^?1) in 120 h, suggesting that the tested strain has potential for successful bioprocess for caffeine degradation. Growth kinetics showed substrate inhibition phenomenon. Various substrate inhibition models were fitted to the kinetic data, amongst which the double-exponential (R ² = 0.94), Luong (R ² = 0.92), and Yano and Koga 2 (R ² = 0.94) models were found to be the best. The Luedeking–Piret model showed that caffeine demethylase production kinetics was growth related. This is the first report on production of high levels of caffeine demethylase in batch bioreactor with faster degradation rate and high tolerance to caffeine, hence clearly suggesting that Pseudomonas sp. used in this study is a potential biocatalyst for industrial decaffeination.     

Interactions between fungal growth,substrate utilization,and enzyme production during solid substrate cultivation of Phanerochaete chrysosporium on cotton stalks

Fungal pretreatment, using lignin-degrading microorganisms to improve lignocellulosic feedstocks with minimal energy input, is a potential alternative to physiochemical pretreatment methods. Identifying the kinetics for fungal pretreatment during solid substrate cultivation is needed to help establish the processing conditions for effective scale up of this technology. In this study, a set of mathematical models were proposed for describing the interactions between holocellulose consumption, lignin degradation, cellulase, ligninolytic enzyme, and the growth of Phanerochaete chrysosporium during a 14 day fungal pretreatment process. Model parameters were estimated and validated by the System Biology Toolbox in MatLab. Developed models provided sufficiently accurate predictions for fungal growth (R ² = 0.97), holocellulose consumption (R ² = 0.97), lignin degradation (R ² = 0.93) and ligninolytic enzyme production (R ² = 0.92), and fair prediction for cellulase production (R ² = 0.61). The models provide valuable information for understanding the interactive mechanisms in biological systems as well as for fungal pretreatment process scale up and improvement.     

A two-step fermentation of distillers’ grains using Trichoderma viride and Rhodopseudomonas palustris for fish feed

It is important to provide added value or to make full use of the co-product of grains from ethanol production. In order to convert distillers’ grains into a high-quality feed, the Trichoderma viride and Rhodopseudomonas palustris fermentation were combined and investigated in this study. The T. viride fermentation was carried out in an aerobic fermentation installation in favoring of the growth of the fungi and the degradation of the cellulose, and then the fermentation of R. palustris was performed to increase the content of protein with an anaerobic installation. After the two step fermentations, the true protein content of dried distiller’ grains increased from 11.4 to 33.6 % (w/w) (the content of crude protein from 14.5 to 39.7 %), the crude fiber content decreased from 21.3 to 7.6 % (w/w), the crude fat content increased from 5.5 to 7.9 % (w/w), the crude ash decreased from 14.6 to 10.2 % (w/w), the total phosphorus content increased from 0.4 to 1.2 % (w/w), and the water content was 11.8 % (w/w). The dried and fermented grains contain the R. palustris viable count of 5.3 × 10¹¹ CFU/g dry matter. The results may support a new application of an active photosynthetic bacteria fish feed in fisheries industry and offer a reference for the further study of lignocellulosic materials as raw materials converting into high-quality feed.     

Microbial detoxification of pathotoxin produced by spot blotch pathogen Bipolaris sorokiniana infecting wheat

Bipolaris sorokiniana causes spot blotch in wheat and barley. The pathogen produces toxin (BS-toxin), which is a sesquiterpenoid belonging to eremophilane family. Isolates of Trichoderma spp., Chaetomium globosum and Pseudomonas fluorescens were tested for detoxification of BS-toxin amended in semi-synthetic medium at different concentrations. All the antagonists showed mycelial growth in toxin amended medium but their growth was less in comparison to growth in normal medium. The growth of biocontrol agents decreased with increasing concentration of toxin. Two isolates of C. globosum (Cg1 and Cg2), T.viride (TV5-2) and Pseudomonas fluorescens produced 4.9, 2.9, 3.6 g mycelium and 5.5 × 10⁵ cfu /ml, respectively exhibiting 50% or less reduction in growth in BS-toxin amended medium at 1,000 ppm concentration. The biocontrol agents also reduced the severity of toxin-induced symptoms and electrolyte leakage from the wheat leaf tissues. Among the microbes tested, maximum reduction in electrolyte leakage was observed in C. globosum (Cg2) treated toxin samples. The spectral analysis also showed a remarkable decrease in optical density of Cg2 treated toxin at 294 nm. High Performance Liquid Chromatography (HPLC) analysis showed almost complete degradation of BS-toxin in C. globosum (Cg2) treated samples.     

Large-scale bioreactor production of the herbicide-degrading Aminobacter sp. strain MSH1

The Aminobacter sp. strain MSH1 has potential for pesticide bioremediation because it degrades the herbicide metabolite 2,6-dichlorobenzamide (BAM). Production of the BAM-degrading bacterium using aerobic bioreactor fermentation was investigated. A mineral salt medium limited for carbon and with an element composition similar to the strain was generated. The optimal pH and temperature for strain growth were determined using shaker flasks and verified in bioreactors. Glucose, fructose, and glycerol were suitable carbon sources for MSH1 (μ?=?0.1 h^?1); slower growth was observed on succinate and acetic acid (μ?=?0.01 h^?1). Standard conditions for growth of the MSH1 strain were defined at pH 7 and 25 °C, with glucose as the carbon source. In bioreactors (1 and 5 L), the specific growth rate of MSH1 increased from μ?=?0.1 h^?1 on traditional mineral salt medium to μ?=?0.18 h^?1 on the optimized mineral salt medium. The biomass yield under standard conditions was 0.47 g dry weight biomass/g glucose consumed. An investigation of the catabolic capacity of MSH1 cells harvested in exponential and stationary growth phases showed a degradation activity per cell of about 3?×?10^?9 μg BAM h^?1. Thus, fast, efficient, large-scale production of herbicide-degrading Aminobacter was possible, bringing the use of this bacterium in bioaugmentation field remediation closer to reality.     

Inorganic carbon and pH effect on growth and lipid productivity of Tetraselmis suecica and Chlorella sp (Chlorophyta) grown outdoors in bag photobioreactors

There has been considerable interest in cultivation of green microalgae (Chlorophyta) as a source of lipid that can alternatively be converted to biodiesel. However, almost all mass cultures of algae are carbon-limited. Therefore, to reach a high biomass and oil productivities, the ideal selected microalgae will most likely need a source of inorganic carbon. Here, growth and lipid productivities of Tetraselmis suecica CS-187 and Chlorella sp were tested under various ranges of pH and different sources of inorganic carbon (untreated flue gas from coal-fired power plant, pure industrial CO₂, pH-adjusted using HCl and sodium bicarbonate). Biomass and lipid productivities were highest at pH 7.5 (320?±?29.9 mg biomass L^?1 day^?1and 92?±?13.1 mg lipid L^?1 day^?1) and pH 7 (407?±?5.5 mg biomass L^?1 day^?1 and 99?±?17.2 mg lipid L^?1 day^?1) for T. suecica CS-187 and Chlorella sp, respectively. In general, biomass and lipid productivities were pH 7.5?>?pH 7?>?pH 8?>?pH 6.5 and pH 7?>?pH 7.5?=?pH 8?>?pH 6.5?>?pH 6?>?pH 5.5 for T. suecica CS-187 and Chlorella sp, respectively. The effect of various inorganic carbon on growth and productivities of T. suecica (regulated at pH?=?7.5) and Chlorella sp (regulated at pH?=?7) grown in bag photobioreactors was also examined outdoor at the International Power Hazelwood, Gippsland, Victoria, Australia. The highest biomass and lipid productivities of T. suecica (51.45?±?2.67 mg biomass L^?1 day^?1 and 14.8?±?2.46 mg lipid L^?1 day^?1) and Chlorella sp (60.00?±?2.4 mg biomass L^?1 day^?1 and 13.70?±?1.35 mg lipid L^?1 day^?1) were achieved when grown using CO₂ as inorganic carbon source. No significant differences were found between CO₂ and flue gas biomass and lipid productivities. While grown using CO₂ and flue gas, biomass productivities were 10, 13 and 18 %, and 7, 14 and 19 % higher than NaHCO₃, HCl and unregulated pH for T. suecica and Chlorella sp, respectively. Addition of inorganic carbon increased specific growth rate and lipid content but reduced biomass yield and cell weight of T. suecica. Addition of inorganic carbon increased yield but did not change specific growth rate, cell weight or content of the cell weight of Chlorella sp. Both strains showed significantly higher maximum quantum yield (F_v/F_m) when grown under optimum pH.     

Optimization of culture conditions for the direct regeneration of Kappaphycus alvarezii (Rhodophyta,Solieriaceae)

Cultivation of seaweeds on a commercial scale requires a large number of propagules with desirable phenotypic traits which include high growth rates and resistance to diseases. Seaweed micropropagation can be considered as one of the best methods to provide a large amount of seedlings for commercial cultivation. This study was carried out to optimize the parameters known to affect the growth of Kappaphycus alvarezii in vitro and subsequently improve the production of seedlings through micropropagation. Suitability of media, concentration of phytoregulators, types and concentration of fertilizers, culture density, light intensity, interval of aeration activity, salinity, and pH were found to be critical factors for the growth of K. alvarezii. The optimum condition for direct regeneration of K. alvarezii in a culture vessel was found to be cultivation of explants in Provasoli's enriched seawater (PES) media supplemented with 2.5 mg L^?1 6-benzylaminopurine (BAP), 1.0 mg L^?1 indole-3-acetic acid (IAA), and 3.0 mg L^?1 natural seaweed extract (NSE) with culture density of 0.4 %?w/v, under light intensity of 75 μmol photons m^?2 s^?1, continuous aeration of 30.0 L h^?1, salinity of 30.0 ppt, and pH 7.5. An airlift photobioreactor was constructed for the mass propagation of K. alvarezii explants with optimum culture conditions obtained from the study. The optimum growth rates of the K. alvarezii explants in culture vessels (5.5 % day^?1) and photobioreactor (6.5 % day^?1) were found to be higher than the growth rate observed in field trials in the open sea (3.5 % day^?1). The information compiled during the course of this study will be of utility to commercial seaweed cultivators.     

Enrichment of biologically active 18-β glycyrrhetinic acid in Glycyrrhiza glabra root by solid state fermentation

This paper describes the in situ bioconversion of glycyrrhizin of Glycyrrhiza glabra root to 18-β glycyrrhetinic acid by solid state fermentation. Fermentation was carried out with two different fungal strains, Penicillium chrysogenum and Rhizopus oryzae. The solid state fermentation was carried out under stationary state and under rotating state. Penicillium chrysogenum is a better producer of 18-β glycyrrhetinic acid than Rhizopus oryzae. The induced P. chrysogenum seed culture produces higher 18-β glycyrrhetinic acid with 2.955 mg g^?1 and maximum β-glucuronidase activity of 3,583.8 U ml^?1 under stationary solid state fermentation. The mycelium growth and bioconversion rate is highest at pH of 5.5 and 4.5, respectively. G. glabra root supplemented with a solution of dextrose 9 g l^?1, MnSO₄?·?H₂O 3 g l^?1 and (NH₄)₂SO₄ 0.540 g l^?1 produces 48.580 mg of 18-β glycyrrhetinic acid per gram of G. glabra root, i.e. 86.74 % bioconversion by P. chrysogenum in 96 h under stationary state solid state fermentation.     

Linking root traits and competitive success in grassland species

Background and aims

Measures of phosphorus (P) in roots recovered from soil underestimate total P accumulation below-ground by crop species since they do not account for P in unrecovered (e.g., fine) root materials. ³³P-labelling of plant root systems may allow more accurate estimation of below-ground P input by plants.

Methods

Using a stem wick-feeding technique ³³P-labelled phosphoric acid was fed in situ to canola (Brassica napus) and lupin (Lupinus angustifolius) grown in sand or loam soils in sealed pots.

Results

Recovery of ³³P was 93 % in the plant-soil system and 7 % was sorbed to the wick. Significantly more ³³P was allocated below-ground than to shoots for both species with 59–90 % of ³³P measured in recovered roots plus bulk and rhizosphere soil. ³³P in recovered roots was higher in canola than lupin regardless of soil type. The proportion of ³³P detected in soil was greater for lupin than canola grown in sand and loam (37 and 73 % lupin, 20 and 23 % canola, respectively). Estimated total below-ground P accumulation by both species was at least twice that of recovered root P and was a greater proportion of total plant P for lupin than canola.

Conclusion

Labelling roots using ³³P via stem feeding can empower quantitative estimates of total below-ground plant P and root dry matter accumulation which can improve our understanding of P distribution in soil-plant systems.

     

The effect of rumen ciliates on chitinolytic activity,chitin content and the number of fungal zoospores in the rumen fluid of sheep

The objective of this study was to investigate the effect of selected protozoa on the degradation and concentration of chitin and the numbers of fungal zoospores in the rumen fluid of sheep. Three adult ewes were fed a hay-concentrate diet, defaunated, then monofaunated with Entodinium caudatum or Diploplastron affine alone and refaunated with natural rumen fauna. The average density of the protozoa population varied from 6.1 · 10⁴ (D. affine) to 42.2 · 10⁴ cells/ml rumen fluid (natural rumen fauna). The inoculation of protozoa in the rumen of defaunated sheep increased the total activity of chitinolytic enzymes from 2.9 to 3.6 μmol N-acetylglucosamine/g dry matter (DM) of rumen fluid per min, the chitin concentration from 6.3 to 7.2 mg/g DM of rumen fluid and the number of fungal zoospores from 8.1 to 10.9 · 10⁵ cells/ml rumen fluid. All examined indices showed diurnal variations. Ciliate population density was highest immediately prior to feeding and lowest at 4 h thereafter. The opposite effects were observed for the numbers of fungal zoospores, the chitin concentration and chitinolytic activity. Furthermore, it was found that chitin from zoospores may account for up to 95% of total microbial chitin in the rumen fluid of sheep. In summary, the examined ciliate species showed the ability of chitin degradation as well as a positive influence on the development of the ruminal fungal population.     

The influence of goethite and gibbsite on soluble nutrient dynamics and microbial community composition

Iron and aluminum (oxyhydr)oxides are ubiquitous in the soil environment and have the potential to strongly affect the properties of dissolved organic matter. We examined the effect of oxide surfaces on soluble nutrient dynamics and microbial community composition using an incubation of forest floor material in the presence of (1) goethite and quartz, (2) gibbsite and quartz, and (3) quartz surfaces. Forest floor material was incubated over a period of 154 days. Aqueous extracts of the incubations were harvested on days 5, 10, 20, 30, 60, 90, and 154, and concentrations of P, N, PO₄ ^3?, NO₂ ^?, NO₃ ^?, and organic C were measured in the solutions. Microbial community composition was examined through pyrosequencing of bacterial and fungal small subunit ribosomal RNA genes on selected dates throughout the incubation. Results indicated that oxide surfaces exerted strong control on soluble nutrient dynamics and on the composition of the decomposer microbial community, while possibly having a small impact on system-level respiration. Goethite and gibbsite surfaces showed preferential adsorption of P-containing and high molar mass organic solutes, but not of N-containing compounds. On average, organic C concentrations were significantly lower in water extractable organic matter (WEOM) solutions from oxide treatments than from the control treatment (P = 0.0037). Microbial community composition varied both among treatments and with increasing time of incubation. Variation in bacterial and fungal community composition exhibited strong-to-moderate correlation with length of incubation, and several WEOM physiochemical characteristics including apparent (weight averaged) molar mass, pH and electrical conductivity. Additionally, variation in bacterial community composition among treatments was correlated with total P (r = 0.60, P < 0.0001), PO₄ ^3? (r = 0.79, P < 0.0001), and organic C (r = 0.36, P = 0.015) concentrations; while variation in fungal communities was correlated with organic C concentrations (r = ?0.48, P = 0.0008) but not with phosphorus concentrations. The relatively small impact of oxide surfaces on system-level microbial respiration of organic matter despite their significant effects on microbial community composition and WEOM dynamics lends additional support to the theory of microbial functional redundancy.     

Polyhydroxyalkanoate biosynthesis and simultaneous remotion of organic inhibitors from sugarcane bagasse hydrolysate by Burkholderia sp.

Burkholderia sp. F24, originally isolated from soil, was capable of growth on xylose and removed organic inhibitors present in a hemicellulosic hydrolysate and simultaneously produced poly-3-hydroxybutyrate (P3HB). Using non-detoxified hydrolysate, Burkholderia sp. F24 reached a cell dry weight (CDW) of 6.8 g L^?1, containing 48 % of P3HB and exhibited a volumetric productivity (P_P3HB) of 0.10 g L^?1 h^?1. Poly-3-hydroxybutyrate-co-3-hydroxyvalerate copolymers (P3HB-co-3HV) were produced using xylose and levulinic acid (LA) as carbon sources. In shake flask cultures, the 3HV content in the copolymer increased from 9 to 43 mol% by adding LA from 1.0 to 5.0 g L^?1. In high cell density cultivation using concentrated hemicellulosic hydrolysate F24 reached 25.04 g L^?1 of CDW containing 49 % of P3HB and P_P3HB of 0.28 g L^?1 h^?1. Based on these findings, second-generation ethanol and bioplastics from sugarcane bagasse is proposed.     

Biomass and Total Lipid Content Assessment of Microalgal Cultures Using Near and Short Wave Infrared Spectroscopy

The technique of near and short wave near-infrared spectroscopy was assessed with respect to analysis of dry matter and lipid content of microalgae with potential for biodiesel production. Microalgal culture samples were filtered through GF/C filter papers and spectral measurements of wet and oven dried (60 °C overnight) filter papers over the ranges of 300–1,100 nm and 1,100–2,500 nm were recorded. Partial least square models on culture biomass and lipid content for combined species data were poor in terms of RMSECV, R _CV and the ratio of RMSECV to SD. A single species model for C. vulgaris based on 1,100–2,500 nm spectra of dry filtrate supported a model with RMSECV, R _CV and SDR values of 0.32 g L^?1, 0.955 and 3.38 for biomass and 0.089 g L^?1, 0.874 and 2.06 with lipid, respectively. However, the dry filtrate models on biomass and lipid content performed poorly in the prediction of samples drawn from an independent series of C. vulgaris cultured under N-, P- and Fe-limited growth trial. Thus, while the near-infrared spectroscopy technique has potential for assessment of dry matter and lipid content of microalgal cultures using a dried filtrate sample, further work is required to examine the limits to model robustness.     

Dry matter yield,root traits,and nodule occupancy of lucerne and Caucasian clover when grown in acidic soil with high aluminium concentrations

Aims and methods

Lucerne and Caucasian clover dry matter were measured in response to recommended lime and capital P inputs for six years in an acidic soil in the New Zealand high country. The initial three years of the field experiment indicated successful establishment and persistence of both legumes. Lucerne dry matter (DM) yield was up to 4 t/ha/yr in this period and higher than Caucasian clover yields. However, a lack of persistence of lucerne was apparent from this point forward compared with Caucasian clover which produced 7.7 t DM/ha in Year 6. An experiment using tubes of soil was used to investigate whether differences in root traits, nodulation and nodule occupancy were responsible for the differences observed in field persistence over time.

Results

These showed that when rhizobia inoculant was added, the fine root length of Caucasian clover was unaffected (R² = 0.14) by aluminium (Al) content of the soil. In contrast, fine root growth of lucerne was suppressed (R² = 0.79) by the soil Al content. Nodulation of Caucasian clover was unaffected by soil pH or Al when the rhizobia inoculant was provided which suggests the viability of the commercial genotype ICC148 in this soil with a pH of 5.5 and Al ca. 7 mg/kg soil. For lucerne, the maximum nodulation score of 7.3 occurred with 2 t/ha of lime added (soil pH ca.6, Al ca. 0.3 mg/kg) plus inoculant.

Conclusions

This suggests an Al toxic threshold of <1.0 mg Al/kg soil for effective lucerne nodulation. From the lucerne nodules, eight naturalized strains of Ensifer meliloti were identified. In contrast, only one R. leguminosarum strain was detected in the Caucasian clover nodules. The competition between those rhizobia genotypes may negatively affect the efficiency of biological nitrogen fixation in lucerne. Therefore, the lack of genetic diversity of R. leguminosarum bv. trifolii in New Zealand soils might be an advantage especially if the commercial strain is acid soil tolerant.