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Few investigations have been carried out into the members of the genus Thermoanerobacter for protein and amino acid metabolism compared to carbohydrates, mostly due to the intense interest in bioethanol and biohydrogen in recent decades. The present study investigates the biotechnological potential of Thermoanaerobacter mathranii (DSM 11426) in terms of its ability to produce high-energy alcohols from amino and fatty acids. End product formation from glucose (in the presence and absence of butyrate) as well as from selected proteins and amino acids were analyzed. T. mathranii did not degrade any of the proteins tested to a large extent but degraded several amino acids, namely serine and the branched-chain amino acids (leucine, isoleucine, valine) when cultivated in the presence of thiosulfate. The main end products from the branched-chain amino acids were a mixture of their corresponding branched-chain fatty acids and alcohols, with the strain producing a concentration of the corresponding branched-chain alcohol between 1.0 and 1.7 mM and 8.2–10.9 mM of the corresponding fatty acid. 13C2-labeled leucine revealed that the strains degraded the amino acid in the presence of thiosulfate, producing 3-methyl-1-butyrate, which was then used as an electron acceptor which led to the accumulation of 3-methyl-1-butanol. The strain is highly ethanologenic, producing more than 1.2 mol of ethanol per mol of glucose degraded. The strain was able to reduce volatile fatty acids during glucose fermentation to their corresponding alcohol, further suggesting this strain may be of greater biotechnological value beyond bioethanol production.

The dataset details the fermentation of D-glucose, L-rhamnose, and L-fucose and their end-product formation by the moderate thermophile Clostridium strain AK1 (DSM 18778) as related to the work described in “Propanediol from L-rhamnose using the moderately thermophilic Clostridium strain AK1” [1]. The influence of culture conditions on end product formation from D-glucose and L-rhamnose by AK1 was investigated in batch culture. Strain AK1 was cultivated at initial substrate concentrations varying from 0 to 60 mM and initial pH values varying from 4.5 to 8.5. Additionally different cultivation temperatures (30–65 °C), the influence of liquid-gas phase ratio as well as different phosphate concentrations on growth were investigated. Keywords: Propylene glycol, Deoxy sugar, Methylpentose, Chiral alcohol, Moderate thermophile

The dataset describes the catabolism of the 20 proteogenics amino acids and their end products by Thermoanaerobacter strain AK85 under different electron scavenging conditions with an emphasis on the branched-chain amino acids as reported in Scully and Orlygsson, 2019. Keywords: Thermophiles, Branched-chain amino acids and alcohols, Thiosulfate

Abstract Sustainably produced renewable biomass has the potential to replace fossil-based feedstocks, for generation of biobased fuels and chemicals of industrial interest, in biorefineries. In this context, seaweeds contain a large fraction of carbohydrates that are a promising source for enzymatic and/or microbial biorefinery conversions. The thermoanaerobe Thermoanaerobacterium AK17 is a versatile fermentative bacterium producing ethanol, acetate and lactate from various sugars. In this study, strain AK17 was engineered for more efficient production of ethanol by knocking out the lactate and acetate side-product pathways. This was successfully achieved, but the strain reverted to acetate production by recruiting enzymes from the butyrate pathway. Subsequently this pathway was knocked out and the resultant strain AK17_M6 could produce ethanol close to the maximum theoretical yield (90%), leading to a 1.5-fold increase in production compared to the wild-type strain. Strain AK17 was also shown to successfully ferment brown seaweed hydrolysate from Laminaria digitata to ethanol in a comparatively high yield of 0.45 g/g substrate, with the primary carbon sources for the fermentations being mannitol, laminarin-derived glucose and short laminari-oligosaccharides. As strain AK17 was successfully engineered and has a wide carbohydrate utilization range that includes mannitol from brown seaweed, as well as hexoses and pentoses found in both seaweeds and lignocellulose, the new strain AK17_M6 obtained in this study is an interesting candidate for production of ethanol from both second and third generations biomass.

Thermoanaerobacter species have recently been observed to reduce carboxylic acids to their corresponding alcohols. The present investigation shows that Thermoanaerobacter pseudoethanolicus converts C2–C6 short-chain fatty acids (SCFAs) to their corresponding alcohols in the presence of glucose. The conversion yields varied from 21% of 3-methyl-1-butyrate to 57.9% of 1-pentanoate being converted to their corresponding alcohols. Slightly acidic culture conditions (pH 6.5) was optimal for the reduction. By increasing the initial glucose concentration, an increase in the conversion of SCFAs reduced to their corresponding alcohols was observed. Inhibitory experiments on C2–C8 alcohols showed that C4 and higher alcohols are inhibitory to T. pseudoethanolicus suggesting that other culture modes may be necessary to improve the amount of fatty acids reduced to the analogous alcohol. The reduction of SCFAs to their corresponding alcohols was further demonstrated using 13C-labelled fatty acids and the conversion was followed kinetically. Finally, increased activity of alcohol dehydrogenase (ADH) and aldehyde oxidation activity was observed in cultures of T. pseudoethanolicus grown on glucose as compared to glucose supplemented with either 3-methyl-1-butyrate or pentanoate, using both NADH and NADPH as cofactors, although the presence of the latter showed higher ADH and aldehyde oxidoreductase (ALDH) activity.

The dataset describes the influence of culture conditions on the bioreduction of organic acids by Thermoanaerobacter pseudethanolicus as reported in [1]. The data shows that during glucose fermentation of Thermoanaerobacter pseudethanolicus the reducing equivalents are not only converted to ethanol and hydrogen but also, in the presence of carboxylic acids (C2–C6), to its corresponding alcohol. To maximize the alcohol production produced from their carboxylic acid, several experiments were performed to investigate the effect of various environmental factors (initial glucose concentration, pH, liquid–gas phase ratio, and inhibitory effects of alcohols) on growth. A kinetic experiment of glucose in the absence and presence of selected fatty acids are also presented as are data on selected enzyme activities related to alcohols and aldehydes and a time course study of the reduction of 13C1 labeled butyrate using glucose as a carbon source.

The dataset depicts the catabolism of branched-chain amino acids by Thermoanaerobacter pseudethanolicus in the presence of thiosulfate under different culture conditions. The results reveal that the strain can degrade all three branched-chain amino acids resulting in the production of their corresponding branched-chain fatty acids and branched-chain alcohols with the fatty acids always being the dominant product. The highest amounts of 2-methyl-1-butanol from isoleucine were at pH 6.5, liquid-gas ratio of 0.98, and at 20 mM thiosulfate concentration. A kinetic experiment of the branched-chain amino acids was done in the presence of thiosulfate as are data on selected enzyme activities related to alcohols and aldehydes. Finally, an NMR study using 13C1 methyl-1-butyrate during the degradation of leucine in the presence of thiosulfate was done to prove that the 13C1-methyl-1-butanol was indeed from its corresponding fatty acid.

Seaweeds (macroalgae) are gaining attention as potential sustainable feedstock for the production of fuels and chemicals. This comparative study focuses on the characterization of the microbial production of alcohols from fermentable carbohydrates in the hydrolysate of the macroalgae Laminaria digitata as raw material. The potential of a hydrolysate as a carbon source for the production of selected alcohols was tested, using three physiologically different fermentative microbes, in two main types of processes. For the production of ethanol, Saccharomyces cerevisiae was used as a benchmark microorganism and compared with the strictly anaerobic thermophile Thermoanaerobacterium strain AK17. For mixed production of acetone/isopropanol, butanol, and ethanol (A/IBE), three strictly anaerobic Clostridium strains were compared. All strains grew well on the hydrolysate, and toxicity constraints were not observed, but fermentation performance and product profiles were shown to be both condition- and strain-specific. S. cerevisiae utilized only glucose for ethanol formation, while strain AK17 utilized glucose, mannitol, and parts of the glucan oligosaccharides. The clostridia strains tested showed different nutrient requirements, and were able to utilize glucan, mannitol, and organic acids in the hydrolysate. The novelty of this study embodies the application of different inoculates for fermenting a common brown seaweed found in the northern Atlantic Ocean. It provides important information on the fermentation properties of different microorganisms and pinpoints the value of carbon source utilization when selecting microbes for efficient bioconversion into biofuel and chemical products of interest.

Thermoanaerobacter strains have recently gained interest because of their ability to convert short chain fatty acids to alcohols using actively growing cells. Thermoanaerobacter thermohydrosulfuricus strain AK152 was physiologically investigated for its ethanol and other alcohol formation. The temperature and pH optimum of the strain was 70 °C and pH 7.0 and the strain degraded a variety of compounds present in lignocellulosic biomass like monosaccharides, disaccharides, and starch. The strain is highly ethanologenic, producing up to 86% of the theoretical ethanol yield form hexoses. Strain AK152 was inhibited by relatively low initial substrate (30 mM) concentration, leading to inefficient degradation of glucose and levelling up of all end-product formation. The present study shows that the strain produces alcohols from most of the tested carboxylic acids, with the highest yields for propionate conversion to propanol (40.7%) with kinetic studies demonstrating that the maximum conversion happens within the first 48 h of fermentation. Various physiological tests were performed to maximize the acid conversion to the alcohol which reveals that the optimum pH for propionate conversion is pH 6.7 which affords a 57.3% conversion. Kinetic studies reveal that propionate conversion is rapid, achieving a maximum conversion within the first 48 h of fermentation. Finally, by using 13C NMR, it was shown that the addition of propionate indeed converted to propanol.

Mannitol-containing macro algae biomass, such as Ascophyllum nodosum and Laminaria digitata, are a potential feedstock for the production of biofuels such as bioethanol. The purpose of this work was to evaluate the ability of thermophilic anaerobes within Class Clostridia to ferment mannitol and mannitol-containing algal extracts. Screening of the type strains of six genera, Caldanaerobius, Caldanaerobacter, Caldicellulosiruptor, Thermoanaerobacter, Thermobrachium, and Thermoanaerobacterium) was conducted on 20 mM mannitol and revealed that 11 of 41 strains could utilize mannitol with ethanol being the dominant end-product. Mannitol utilization seems to be most common within the genus of Thermoanaerobacter (7 of 16 strains) with yields up to 88% of the theoretical yield in the case of Thermoanaerobacter pseudoethanolicus. Six selected mannitol-degrading strains (all Thermoanaerobacter species) were grown on mannitol extracts prepared from A. nodosum and L. digitata. Five of the strains produced similar amounts of ethanol as compared with ethanol yields from mannitol only. Finally, T. pseudoethanolicus was kinetically monitored using mannitol and mannitol extracts made from two macro algae species, A. nodosum and L. digitata for end-product formation.

There is an increased interest in using thermophilic bacteria for the production of bioethanol from complex lignocellulosic biomass due to their higher operating temperatures and broad substrate range. This review focuses upon the main genera of thermophilic anaerobes known to produce ethanol, their physiology, and the relevance of various environmental factors on ethanol yields including the partial pressure of hydrogen, ethanol tolerance, pH and substrate inhibition. Additionally, recent development in evolutionary adaptation and genetic engineering of thermophilic bacteria is highlighted. Recent developments in advanced process techniques used for ethanol production are reviewed with an emphasis on the advantages of using thermophilic bacteria in process strategies including separate saccharification and fermentation, simultaneous saccharification and fermentation (SSF), and consolidated bioprocessing (CBP).