Microbial lipid production by oleaginous Rhodococci cultured in lignocellulosic autohydrolysates.

Metabolic synthesis of single cell oils (SCOs) for biodiesel application by heterotrophic oleaginous microorganisms is being hampered by the high cost of culture media. This study investigated the possibility of using loblolly pine and sweetgum autohydrolysates as economic feedstocks for microbial lipid production by oleaginous Rhodococcus opacus (R. opacus) PD630 and DSM 1069. Results revealed that when the substrates were detoxified by the removal of inhibitors (such as HMF-hydroxymethyl-furfural), the two strains exhibited viable growth patterns after a short adaptation/lag phase. R. opacus PD630 accumulated as much as 28.6 % of its cell dry weight (CDW) in lipids while growing on detoxified sweetgum autohydrolysate (DSAH) that translates to 0.25 g/l lipid yield. The accumulation of SCOs reached the level of oleagenicity in DSM 1069 cells (28.3 % of CDW) as well, while being cultured on detoxified pine autohydrolysate (DPAH), with the maximum lipid yield of 0.31 g/l. The composition of the obtained microbial oils varied depending on the substrates provided. These results indicate that lignocellulosic autohydrolysates can be used as low-cost fermentation substrates for microbial lipid production by wild-type R. opacus species. Consequently, the variety of applications for aqueous liquors from lignocellulosic pretreatment has been expanded, allowing for the further optimization of the integrated biorefinery.

Pyrolysis oil-based lipid production as biodiesel feedstock by Rhodococcus opacus.

Light oil from pyrolysis, which accounts for ∼10 % carbon yield of the starting biomass, is a complex aqueous product that is difficult to utilize and usually discarded. This work presents the feasibility of light oil as a sole carbon source to support the growth of Rhodococcus opacus (R. opacus) that in turn accumulate triacylglycerols as biodiesel feedstock. Two types of bacteria (R. opacus PD630 and DSM 1069) were selected in this study. Research results showed that after short adaption periods both strains can grow well on this complex carbon source, as proved by the consumption of oligomers and monomers in light oil. Lipid content by R. opacus PD630 and DSM 1069 was observed up to 25.8 % and 22.0 % of cell dry weight, respectively. Palmitic and stearic acids were found to be the predominant fatty acids in these bacterial cells. In addition, the light oil-based lipid production can be enhanced by reducing the pH value from 7 to 4, especially in case of DSM 1069.

Polymerization of Kraft lignin via ultrasonication for high-molecular-weight applications.

Kraft lignin is an inexpensive and abundant byproduct of pulp mills that can be used in the synthesis of adhesives and carbon fibers along with energy production. Some of these material applications favor the utilization of high molecular weight (HMW) lignin. This study investigates the use of ultrasonics as a means to increase the degree of polymerization (DP) of highly purified Kraft lignin. Treated samples were characterized by gel permeation chromatography (GPC), Fourier transform infrared (FTIR) spectroscopy, (13)C and (31)P nuclear magnetic resonance (NMR). After 15 min of sustained cavitation, ultrasonicated lignin generated a high molecular-weight fraction (~35%) that had a weight-average molecular weight (Mw) over 450-fold greater than the initial Kraft lignin sample. (13)C-NMR and (31)P-NMR analysis indicated that the highly-polymerized fraction was enriched with C5 condensed phenolic structures.

Bioconversion of lignin model compounds with oleaginous Rhodococci.

Although economically efficient biomass conversion depends on the utilization of the complete cell wall (biorefinery concept), including polysaccharides and lignin, current biofuels research concentrate mostly on cellulose conversion, while lignin is viewed as a side-product that is used primarily as a thermal resource. Microbiological conversion of lignin is almost exclusive to fungi, usually resulting in increased cell mass and lignolytic enzymes. Some bacteria can also degrade lignin-related compounds using the ß-ketoadipate pathway; for example, Rhodococcus opacus DSM 1069 can degrade coniferyl alcohol and grow on it as sole carbon source. Moreover, this strain belongs to the actinomycetes group that is also known for oleaginous species with lipid accumulation over 20%. Present work shows that R. opacus DSM 1069 and PD630 strains under nitrogen limiting conditions can convert lignin model compounds into triacylglycerols, also known as neutral lipids. 4-Hydroxybenzoic and vanillic acid lignin model compounds were used as sole carbon sources, and after brief adaptation periods, the cells not only began growing but accumulated lipids to the level of oleaginicity. These lipids were extracted for transesterification and analysis of fatty acid methyl esters showed good composition for biodiesel applications with no aromatics. Furthermore, the two strains showed distinct substrate metabolism and product profiles.

Challenges of the utilization of wood polymers: how can they be overcome?

Diminishing fossil fuel resources as well as growing environmental and energy security concerns, in parallel with growing demands on raw materials and energy, have intensified global efforts to utilize wood biopolymers as a renewable resource to produce biofuels and biomaterials. Wood is one of the most abundant biopolymer composites on earth that can be converted into biofuels as well as used as a platform to produce bio-based materials. The major biopolymers in wood are cellulose, hemicelluloses, and lignin which account for >90% of dry weight. These polymers are generally associated with each other in wood cell walls resulting in an intricate and dynamic cell wall structure. This mini-review provides an overview of major wood biopolymers, their structure, and recent developments in their utilization to develop biofuels. Advances in genetic modifications to overcome the recalcitrance of woody biomass for biofuels are discussed and point to a promising future.

Lipids from heterotrophic microbes: advances in metabolism research.

Heterotrophic oleaginous microorganisms are capable of producing over 20% of their weight in single cell oils (SCOs) composed of triacylglycerols (TAGs). These TAGs contain fatty acids, such as palmitic, stearic and oleic acids, that are well-suited for biodiesel applications. Although some of these microbes are able to accumulate SCOs while growing on inexpensive agro-industrial biomass, the competition with plant oil resources means that a significant increase in productivity is desired. The present review aims to summarize recent details in lipid metabolism research and engineering (e.g. direct fatty acid ethyl ester production), as well as culture condition optimization and innovations, such as solid-state or semi-solid-state fermentation, that can all contribute to higher productivity and further advancement of the field.