**8. Conclusions and perspectives**

Oleaginous fungi, and particularly yeasts, are very efficient in the accumulation of intracellular TAG and it is expected that they will be exploited by the biofuel industry in the future. Nonetheless, the costs of microbial lipids are still too high in order to compete with plant oils for biodiesel manufacturing. Cheap carbon sources have necessarily to be used as carbon sources for the cultivation of these microorganisms and the performance of the bioprocess has to be further improved in terms of both the yield and the productivity. The exploration of the natural biodiversity is a promising strategy to identify novel oleaginous species that assimilate and get fat on agro-industrial residues, particularly the lingo-cellulosic biomass and crude glycerol from biodiesel industry. Further approaches combining genomic, transcriptomic, metabolomics, and lipidomic techniques will undoubtedly provide deeper information of lipid production by oleaginous fungi. A metabolic engineering approach is very promising, but it is still precluded for the most oleaginous species, for which genome disclosure has not been accomplished and genetic tools are not available yet.

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**5** 

*USA* 

**Microbial Biodiesel** 

Jianguo Zhang and Bo Hu *University of Minnesota,* 

**Production - Oil Feedstocks** 

**Produced from Microbial Cell Cultivations** 

Crude oil price has increased to over \$100 per barrel, which is causing serious negative impact on the global and national economy. In 2005, the United States produced 8.3 million bbl/day, but consumed 20.8 million bbl/day, the balance of which was imported from other countries. For these fossil fuels, the U.S. used about 138 billion gallons of gasoline in 2006, accounting for about 44 percent of the world's gasoline consumption (EarthTrends, 2008). The annual U.S. usage of jet fuel was 21 billion gallons in 2006 (Energy Information Administration, 2008). The U.S. annual consumption of diesel fuel in 2006 was about 50 billion gallons (Energy Information Administration, 2008). Massive consumption of fossil fuels has already caused serious concern over global warming caused by greenhouse

Biofuel offers an alternative to fossil fuels. It provides several benefits, such as alleviation from foreign oil dependence, carbon neutral process without greenhouse emission, and profits to local farmers. Bioethanol production from starch and lignocellulosic materials is a kind of an alternative to fossil fuels. It can be blended with gasoline in varying quantities up to pure ethanol (E100). The first generation of ethanol biofuel has been massively commercialized and dominated by the U.S. and Brazil. Fuel ethanol in the U.S. is primarily produced from corn, while Brazilian ethanol is produced mainly from sugarcane. These raw materials are in direct competition with human diet or the land to produce food, which triggers the controversy of food versus fuel. The second generation of ethanol is proposed to be produced from lignocellulosic biomass, which can be obtained from agricultural residue or other woody and herbal biomass from marginal land. Intense scientific research has been carried out over the past decade, focusing on this route in order to decrease the overall

Another approach for alternative biofuels is biodiesel. The most common type of biodiesel is the methyl esters of fatty acid (FAME), obtained by transesterification of lipid with methanol or ethanol. It can be used in pure form (B100) or may be blended with fossil diesel at any rate. The commonly used biodiesel is B99 because 1% of fossil fuel is applied to

process cost and this process is gradually focusing on commercialization.

**2. Biodiesel and current feedstocks** 

**2.1 Biodiesel production** 

**1. Introduction** 

gases.

