Microbial renewable feedstock utilization: a substrate-oriented approach

Abstract

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.

Keywords: bioprocessing; filamentous fungi; industrial biotechnology; second generation feedstock.

Figure 1

Substrate utilization and product production performance of P. stipitis on sugar cane bagasse (AH), wheat straw (AH), corn stover (AH), glycerol, wheat straw (EH), sugar cane bagasse (EH) and willow wood (AH).

Figure 2

Substrate utilization and product production performance of E. coli on sugar cane bagasse (AH), wheat straw (AH), corn straw (AH), sugar cane bagasse (EH), wheat straw (EH).

Figure 3

Substrate utilization and product production performance of S. cerevisiae on sugar cane bagasse (AH), wheat straw (AH), corn straw (AH), sugar cane bagasse (EH), wheat straw (EH) and willow wood (AH).

Figure 4

Substrate utilization and product production performance of A. niger on sugar cane bagasse (AH), wheat straw (AH), corn stover (AH), glycerol, wheat straw (EH), sugar cane bagasse (EH) and willow wood (AH).

Figure 5

Substrate utilization and product production performance of T. reesei on sugar cane bagasse (AH), wheat straw (AH), corn stover (AH), glycerol, wheat straw (EH), sugar cane bagasse (EH) and willow wood (AH).

Figure 6

Substrate utilization and product production performance of C. glutamicum on sugar cane bagasse (AH), wheat straw (AH), corn stover (AH), wheat straw (EH), sugar cane bagasse (EH).

Figure 7

Substrate utilization and product production performance of P. stipitis, A. niger and T. reesei on crude glycerol.

Figure 8

Conversion of HMF and furfural in hydrolyzed corn stover fermentations of A. niger strain N402 at pH 4.5.