Characterisation of a Novel Nitrogen-Fixing, Polyhydroxyalkanoate-Producing Bacterium, Novosphingobium Nitrogenifigens Y88T

<p>The novel sphingomonad Novosphingobium nitrogenifigens Y88T (Y88T) is an obligate aerobe able to grow in nutrient-imbalanced environments where nitrogen is naturally limiting, but carbon is found in abundance. Due to its ability to fix atmospheric nitrogen and produce the bioplastic polyhydroxyalkanoate (PHA), Y88T is well-suited for growth in a nitrogenlimited but carbon-enriched environment. Because of these metabolic abilities, Y88T is of interest as a model organism for PHA production unconstrained by nitrogen-limiting conditions. Growth profiles and PHA production profiles were determined for Y88T under conditions of carbon enrichment, nitrogen sufficiency and depletion to investigate carbon and nitrogen utilisation as well as PHA production in this organism. Also, since the nitrogenase enzyme required for nitrogen fixation is oxygen labile, the effect of DO concentration and the relationship between aerobic metabolism and the nitrogen-fixing and PHA-producing abilities of Y88T was investigated. This study demonstrated: that glucose is the preferred growth substrate for Y88T; that no direct relationship exists between nitrogen fixation and PHB accumulation in Y88T; that Y88T can reliably produce in excess of 80 % of its dry weight as polyhydroxybutyrate (PHB), a type of PHA, from glucose under nitrogenlimiting conditions. Proteomic signatures were determined for the various physiological responses of Y88T to growth, nitrogen utilisation, PHB production and exposure to different levels of DO. More than 250 unique proteins, including the core nitrogen-fixation, PHB-synthetic and glycolytic proteins were identified. Y88T apparently converts glucose to PHB via three interrelated glucose catabolic pathways and proteins likely involved in these pathways were identified. This study revealed that, regardless of growth conditions and despite decreased abundance of the Y88T nitrogenase enzyme, growth and PHB synthesis were not inhibited at DOhigh concentrations. Proteomic characterisation of the Y88T phasin, a PHA granule-associated protein, iii identified an amino-terminal, low complexity alanine and proline rich segment found only in other sphingomonads. The expression level of the Y88T phasin correlated well with PHB yields, suggesting the use of this protein as a biomarker to optimise PHB yield in a production environment. Y88T has the potential to be a useful production strain in pure culture, utilising its natural and robust propensity to metabolise glucose to preferentially produce PHB. Targets for biotechnological improvement and the potential for application of Y88T to biofuel production are discussed.</p>

Characterisation of a Novel Nitrogen-Fixing, Polyhydroxyalkanoate-Producing Bacterium, Novosphingobium Nitrogenifigens Y88T

<p>The novel sphingomonad Novosphingobium nitrogenifigens Y88T (Y88T) is an obligate aerobe able to grow in nutrient-imbalanced environments where nitrogen is naturally limiting, but carbon is found in abundance. Due to its ability to fix atmospheric nitrogen and produce the bioplastic polyhydroxyalkanoate (PHA), Y88T is well-suited for growth in a nitrogenlimited but carbon-enriched environment. Because of these metabolic abilities, Y88T is of interest as a model organism for PHA production unconstrained by nitrogen-limiting conditions. Growth profiles and PHA production profiles were determined for Y88T under conditions of carbon enrichment, nitrogen sufficiency and depletion to investigate carbon and nitrogen utilisation as well as PHA production in this organism. Also, since the nitrogenase enzyme required for nitrogen fixation is oxygen labile, the effect of DO concentration and the relationship between aerobic metabolism and the nitrogen-fixing and PHA-producing abilities of Y88T was investigated. This study demonstrated: that glucose is the preferred growth substrate for Y88T; that no direct relationship exists between nitrogen fixation and PHB accumulation in Y88T; that Y88T can reliably produce in excess of 80 % of its dry weight as polyhydroxybutyrate (PHB), a type of PHA, from glucose under nitrogenlimiting conditions. Proteomic signatures were determined for the various physiological responses of Y88T to growth, nitrogen utilisation, PHB production and exposure to different levels of DO. More than 250 unique proteins, including the core nitrogen-fixation, PHB-synthetic and glycolytic proteins were identified. Y88T apparently converts glucose to PHB via three interrelated glucose catabolic pathways and proteins likely involved in these pathways were identified. This study revealed that, regardless of growth conditions and despite decreased abundance of the Y88T nitrogenase enzyme, growth and PHB synthesis were not inhibited at DOhigh concentrations. Proteomic characterisation of the Y88T phasin, a PHA granule-associated protein, iii identified an amino-terminal, low complexity alanine and proline rich segment found only in other sphingomonads. The expression level of the Y88T phasin correlated well with PHB yields, suggesting the use of this protein as a biomarker to optimise PHB yield in a production environment. Y88T has the potential to be a useful production strain in pure culture, utilising its natural and robust propensity to metabolise glucose to preferentially produce PHB. Targets for biotechnological improvement and the potential for application of Y88T to biofuel production are discussed.</p>

Effects of interspecific competition on crop yield and nitrogen utilisation in maize-soybean intercropping system

Intercropping system plays a crucial role in improving crop yield, nitrogen utilisation efficiency (NUE) and economic benefit. The difference in crop yield and interspecific relationship under different bandwidth and row ratio allocation patterns are still unclear. A field experiment was carried out to explore change regularities between crop yield and interspecific relationships under maize soybean intercropping with different bandwidths and row ratios. The results showed that the yield of intercropped crops was lower than that of the sole crop. The nitrogen accumulation (NA), NUE and nitrogen competition ratio was the highest under the intercropping mode with a bandwidth of 2.0 m, which indicated that this mode was more conducive to the N uptake and utilisation in crops. In all intercropping systems, nitrogen equivalent ratio (NER) and land equivalent ratio (LER) were all greater than one, indicating that intercropping systems were conducive to improving land utilisation efficiency and NUE. Under the same bandwidth pattern, expanding the maize soybean row ratio from 2 : 4 to 3 : 4 was beneficial to the improvement of LER, NER, NUE, crop group yield. In conclusion, it was preferable in the NA, NUE, crop group yield under the system of bandwidth 2.0 m and row ratio 2 : 2, which could be a reference for maize soybean intercropping system.

Proxy Measures and Novel Strategies for Estimating Nitrogen Utilisation Efficiency in Dairy Cattle

The efficiency with which dairy cows convert dietary nitrogen (N) to milk N is generally low (typically 25%). As a result, much of the N consumed is excreted in manure, from which N can be lost to the environment. Therefore there is increasing pressure to reduce N excretion and improve N use efficiency (NUE) on dairy farms. However, assessing N excretion and NUE on farms is difficult, thus the need to develop proximate measures that can provide accurate estimates of nitrogen utilisation. This review examines a number of these proximate measures. While a strong relationship exists between blood urea N and urinary N excretion, blood sampling is an invasive technique unsuitable for regular herd monitoring. Milk urea N (MUN) can be measured non-invasively, and while strong relationships exist between dietary crude protein and MUN, and MUN and urinary N excretion, the technique has limitations. Direct prediction of NUE using mid-infrared analysis of milk has real potential, while techniques such as near-infrared spectroscopy analysis of faeces and manure have received little attention. Similarly, techniques such as nitrogen isotope analysis, nuclear magnetic resonance spectroscopy of urine, and breath ammonia analysis may all offer potential in the future, but much research is still required.