Abstract

Pseudomonas oleovorans and recombinant strains containing the alkane oxidation genes can produce alkane oxidation products in two-liquid phase bioreactor systems. In these bioprocesses the cells, which grow in the aqueous phase, oxidize apolar, non-water soluble substrates. The apolar products typically accumulate in the emulsified apolar phase. We have studied both the bioconversion systems and several downstream processing systems to separate and purify alkanols from these two-liquid phase media. Based on the information generated in these studies, we have now designed bioconversion and downstream processing systems for the production of 1-alkanols from n-alkanes on a 10 kiloton/yr scale, taking the conversion of n-octane to 1-octanol as a model system. Here, we describe overall designs of fed-batch and continuous-fermentation processes for the oxidation of octane to 1-octanol by Pseudomonas oleovorans, and we discuss the economics of these processes. In both systems the two-liquid phase system consists of an apolar phase with hexadecene as the apolar carrier solvent into which n-octane is dissolved, while the cells are present in the aqueous phase. In one system, multiple-batch fermentations are followed by continuous processing of the product from the separated apolar phase. The second system is based on alkane oxidation by continuously growing cultures, again followed by continuous processing of the product. Fewer fermentors were required and a higher space-time-yield was possible for production of 1-octanol in a continuous process. The overall performance of each of these two systems has been modeled with Aspen software. Investment and operating costs were estimated with input from equipment manufacturers and bulk-material suppliers. Based on this study, the production cost of 1-octanol is about 7 US$kg-1 when produced in the fed-batch process, and 8 US$kg-1 when produced continuously. The comparison of upstream and downstream capital costs and production costs showed significantly higher upstream costs for the fed-batch process and slightly higher upstream costs for continuous fermentation. The largest cost contribution was due to variable production costs, mainly resulting from media costs. The organisms used in these systems are P. putida alk+ recombinants which oxidize alkanes, but cannot oxidize the resulting alkanols further. Hence, such cells need a second carbon source, which in these systems is glucose. Although the continuous process is about 10% more expensive than the fed-batch process, improvements to reduce overall cost can be achieved more easily for continuous than for fed-batch fermentation by decreasing the dilution rate while maintaining near constant productivity. Improvements relevant to both processes can be achieved by increasing the biocatalyst performance, which results in improved overall efficiency, decreased capital investment, and hence, decreased production cost. Copyright 1999 John Wiley & Sons, Inc.

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