Discovery of a novel thermostable Zn2+ -dependent alcohol dehydrogenase from Chloroflexus aurantiacus through conserved domains mining.

AIMS: The purpose of the study was to demonstrate feasibility of the Conserved Domains Database (CDD) for identification of novel biocatalysts with desirable properties from a class of well-characterized biocatalysts. METHODS AND RESULTS: The thermostable ADH from Sulfolobus solfataricus with a broad substrate range was applied as a template for the search for novel thermostable ADHs via CDD. From the resulting hits, a putative ADH gene from the thermophilic organism Chloroflexus aurantiacus was cloned and expressed in Escherichia coli. The resulting enzyme was purified and characterized. With a temperature activity optimum of 70°C and a broad substrate spectrum especially for diketones, a versatile new biocatalyst was obtained. CONCLUSIONS: Database-based mining in CDD is a suitable approach to obtain novel biocatalysts with desirable properties. Thereby, the available diversity of similar but not equal enzymes within this class can be increased. SIGNIFICANCE AND IMPACT OF THE STUDY: For industrial applications, there is a demand for larger diversity of similar well-characterized enzymes in order to test them for a given process (biodiversity screening). For fundamental science, the comparison of enzymes with similar function but different sequence can provide insight into structure function relationships or the evolution of enzymes. This study gives a good example on how this demand can be efficiently met.

Comparing centralised and decentralised anaerobic digestion of stillage from a large-scale bioethanol plant to animal feed production.

A comparison of stillage treatment options for large-scale bioethanol plants was based on the data of an existing plant producing approximately 200,000 t/yr of bioethanol and 1,400,000 t/yr of stillage. Animal feed production--the state-of-the-art technology at the plant--was compared to anaerobic digestion. The latter was simulated in two different scenarios: digestion in small-scale biogas plants in the surrounding area versus digestion in a large-scale biogas plant at the bioethanol production site. Emphasis was placed on a holistic simulation balancing chemical parameters and calculating logistic algorithms to compare the efficiency of the stillage treatment solutions. For central anaerobic digestion different digestate handling solutions were considered because of the large amount of digestate. For land application a minimum of 36,000 ha of available agricultural area would be needed and 600,000 m(3) of storage volume. Secondly membrane purification of the digestate was investigated consisting of decanter, microfiltration, and reverse osmosis. As a third option aerobic wastewater treatment of the digestate was discussed. The final outcome was an economic evaluation of the three mentioned stillage treatment options, as a guide to stillage management for operators of large-scale bioethanol plants.