Vincent M. Gray

Microbial utilization of crude glycerol for the production of value-added products

Energy fuels for transportation and electricity generation are mainly derived from finite and declining reserves of fossil hydrocarbons. Fossil hydrocarbons are also used to produce a wide range of organic carbon-based chemical products. The current global dependency on fossil hydrocarbons will not be environmentally or economically sustainable in the long term. Given the future pessimistic prospects regarding the complete dependency on fossil fuels, political and economic incentives to develop carbon neutral and sustainable alternatives to fossil fuels have been increasing throughout the world. For example, interest in biodiesel has undergone a revival in recent times. However, the disposal of crude glycerol contaminated with methanol, salts, and free fatty acids as a by-product of biodiesel production presents an environmental and economic challenge. Although pure glycerol can be utilized in the cosmetics, tobacco, pharmaceutical, and food industries (among others), the industrial purification of crude glycerol is not economically viable. However, crude glycerol could be used as an organic carbon substrate for the production of high-value chemicals such as 1,3-propanediol, organic acids, or polyols. Microorganisms have been employed to produce such high-value chemicals and the objective of this article is to provide an overview of studies on the utilization of crude glycerol by microorganisms for the production of economically valuable products. Glycerol as a by-product of biodiesel production could be used a feedstock for the manufacture of many products that are currently produced by the petroleum-based chemical industry.

Microalgal fatty acid composition: implications for biodiesel quality

The fuel properties of microalgal biodiesel are predicted using published microalgal fatty acid (FA) compositions and predictive fuel models. Biodiesels produced from the microalgae investigated are predicted to have extremely poor oxidative stabilities and the majority also have poor cold-flow properties. The cetane number in most cases is out of specification, but less so than the oxidative stability and cold flow. These findings support the idea that feedstocks rich in monounsaturated fatty acids (MUFAs) are desirable for biodiesel but the composition of the saturated fatty acids (SFAs) is also shown to be of great importance. There is an apparent relationship between algal class and the percentage of FAs represented by MUFA. This potentially allows for the identification of high-MUFA algal classes, or at least provides some basis for researchers to make initial selections of target classes for bioprospecting. Comparisons of FA groups between algal classes also show that the SFAs of Mediophyceae contain significantly higher proportions of C14:0, which is in contrast to the normally abundant C16:0 and the Mediophyceae therefore have better cold-flow characteristics than other classes with similar total SFA contents. Certain particularly promising cases for biodiesel production are presented as species level examples of feedstocks that are close to satisfying the biodiesel standards and to further illustrate the challenges that remain. Variation in FA composition as a response to changes in certain environmental variables forms another important facet to feedstock selection and is briefly considered, with suggestions for further research.

Carbon : nitrogen : phosphorus ratios influence biofilm formation by Enterobacter cloacae and Citrobacter freundii

Aims:  To test the effects of C : N : P ratio modification of a well‐known nutrient medium formulation, the Endo formulation on biofilm formation by Enterobacter cloacae Ecl and Citrobacter freundii Cf1 in both single‐species and binary species biofilms.

Secondary somatic embryogenesis of cassava on picloram supplemented media

The object of this study was to evaluate different strategies for the production of secondary somatic embryos of cassava on picloram-supplemented media. Embryogenically competent calli maintained on double-strength Murashige and Skoog (1962) (MS) medium supplemented with 1 mg l−1 picloram were used as starting material. Secondary embryogenesis from this callus was tested using various basal salt media in either the solid or the liquid state and containing two different concentrations of picloram. Some of the factors effecting the conversion of the embryos into plantlets were also studied. A liquid Schenck and Hildebrand (1972) medium containing 60 g l−1 sucrose and 12 mg l−1 picloram favoured the continual production of a highly embryogenic nodular callus. The normal development of somatic embryos from this tissue was dependant on the use of a picloram-free MS basal salt medium. The embryos were desiccated over a saturated salt solution of K2SO4 (RH 97.5% at 25 °C) and allowed to develop into plantlets on a MS medium containing 0.1 mg l−1 BA. This procedure allowed for the normal elongation of the embryonic hypocotyl and formation of vigorous and viable shoots and roots.

Microbial renewable feedstock utilization: A substrate-oriented approach

Increasingly lignocellulosic biomass hydrolysates are used as the feedstock for industrial fermentations. These biomass hydrolysates consist of complex mixtures of different fermentable sugars, but also contain inhibitors and salts that affect the performance of the product-generating microbes. The performance of six industrially relevant microorganisms, i.e. two bacteria (Escherichia coli and Corynebacterium glutamicum ), two yeasts ( Saccharomyces cerevisiae and Pichia stipitis ) and two fungi ( Aspergillus niger and Trichoderma reesei ) were compared for their ability to utilize and grow on different feedstock hydrolysates (corn stover, wheat straw, sugar cane bagasse and willow wood). Moreover, the ability of the selected hosts to utilize waste glycerol from the biodiesel industry was evaluated. P. stipitis and A. niger were found to be the most versatile and C. glutamicum, and S. cerevisiae were shown to be the least adapted to renewable feedstocks. Clear differences in the utilization of the more abundant carbon sources in these feedstocks were observed between the different species. Moreover, in a species-specific way the production of various metabolites, in particular polyols, alcohols and organic acids was observed during fermentation. Based on the results obtained we conclude that a substrate-oriented instead of the more commonly used product oriented approach towards the selection of a microbial production host will avoid the requirement for extensive metabolic engineering. Instead of introducing multiple substrate utilization and detoxification routes to efficiently utilize lignocellulosic hydrolysates only one biosynthesis route forming the product of interest has to be engineered.

Phosphorus limitation and starvation effects on cell growth and lipid accumulation in Isochrysis galbana U4 for biodiesel production

The effect of varying levels of phosphorus (P) on Isochrysis galbana U4 growth, pigmentation and lipid accumulation were investigated. A reduction in the P content to 25% of the recommended level for f/2 medium did not lead to declines in cell growth rates or lipid accumulation levels relative to the cultures maintained on medium supplemented with the normal P dose. Evidence suggesting that the recommended P supply in f/2 exceeds the requirements for maximal algal growth has obvious economic implications for the mass production of I. galbana for biodiesel production. When P supply was in excess this species was also found to accumulate intracellular levels of P that exceeded by up to 6 times its P requirements for growth and cell division. The reduction in P concentration to levels below 25% resulted in P starvation stimulated chlorophyll reductions and carotenoid and lipid accumulation in this species.

Development of Cassava (Manihot esculenta Crantz.) somatic embryos during culture with abscisic acid and activated charcoal

Summary Secondary cassava somatic embryos were cultured on a sequence of developmental media containing either activated charcoal or abscisic acid. Medium supplementation with activated charcoal had a positive effect on both differentiation and subsequent germination. However, sustained exposure to activated charcoal supplemented media beyond the seventh day of differentiation did not appear to be beneficial. Somatic embryos were desiccated over a saturated salt solution of K 2 SO 4 for 10 days and allowed to develop into plantlets on a MS medium containing 0.4 μmol/L BAP. Embryo germination was significantly affected by the culture medium used prior to the desiccation treatment, with culture on a medium supplemented with ABA at a level of 0.1 μmol/L prior to desiccation significantly enhancing germination. The number of normal germinants produced as judged by morphological criteria were not however, dependant on ABA supplementation of the culture medium prior to desiccation. Culture on a separate medium for the differentiation of embryos, different to the medium used for the maturation of somatic embryos is indicated.

The effect of temperature and effluent recycle rate on hydrogen production by undefined bacterial granules.

Biohydrogen production in an anaerobic fluidized granular bed bioreactor was strongly dependent on temperature and effluent recycle rates. At 45 °C as the effluent recycle rate was increased from 1.3 to 3.5 L/min, the total H₂ output for the bioreactor increased from 10.6 to 43.2 L/h. Volumetric H(2) productivity also increased from 2.1 to 8.7 L H₂/L/h. At 70°C as the effluent recycle was increased from 1.3 to 3.5 L/min, the total H₂ output for the bioreactor increased from 13.8 to 73.8L/h. At 70 °C volumetric H(2) productivities increased from 2.8 to 14.8L H₂/L/h as the effluent recycle rate was increased from 1.3 to 3.5 L/min. At 45 °C % H₂ was 45% and reached 67% at 70 °C. Maximum hydrogen yields at 45 °C were 1.24 and 2.2 mol H₂/mol glucose at 70 °C.

Influence of nitrogen stress on Isochrysis galbana strain U4, a candidate for biodiesel production.

Lipid accumulation has been investigated in numerous microalgal species to assess their potential with respect to biodiesel production. The present work determines the effect of nitrogen stress on physiological and ultrastructural changes in Isochrysis galbana U4. This study is unique in showing the correlations between growth, lipid production, pigmentation and ultrastructural changes in Isochrysis cells undergoing nitrogen starvation. The continuation of algal growth after the complete depletion of external nitrogen was shown to be supported by internal nitrogen stores, possibly in the pyrenoid. Cell growth ceased and lipid accumulation was initiated after the internal store of nitrogen had become exhausted. The depletion of intracellular nitrogen reservoirs to critical thresholds initiated the onset of the stationary phase, a decline in chlorophyll content and the initiation of lipid and carotenoid accumulation. The most notable ultrastructural changes, upon nitrogen stress, were the accumulation of plastidial and cytoplasmic lipid bodies and the dismantling of the chloroplast. The size of the pyrenoid when external nitrogen became depleted was found to decrease significantly, up to four‐fold. This was attributed to the remobilization of nitrogen from Rubisco. The level of expression of heterochromatin was found to increase when cells were nitrogen starved. This is thought to favor long‐term dormancy in this species because aging cells have been noted to recover rapidly when returned to conditions favorable for growth. The observations of this study are consistent with the hypothesis that the responses of Isochrysis cells to nitrogen starvation are regulated by the internal reserves of nitrogen, and the depletion of these reserves is an important trigger for lipid accumulation in this species. The findings of this study also indicate that Isochrysis galbana U4 is a promising candidate for biodiesel lipid production.

Draft Whole-Genome Sequence of Serratia marcescens Strain MCB, Associated with Oscheius sp. MCB (Nematoda: Rhabditidae) Isolated from South Africa

ABSTRACT Here we report on the draft genome sequence of Serratia marcescens strain MCB associated with Oscheius sp. MCB (Nematoda: Rhabditidae) isolated from South African soil. S. marcescens strain MCB has 5,304,212-bp genome size with 4,877 genes and a G+C content of 59.1%.