Survey of microbes in industrial-scale second-generation bioethanol production for better process knowledge and operation.

The microbes present in bioethanol production processes have been previously studied in laboratory-scale experiments, but there is a lack of information on full-scale industrial processes. In this study, the microbial communities of three industrial bioethanol production processes were characterized using several methods. The samples originated from second-generation bioethanol plants that produce fuel ethanol from biowaste, food industry side streams, or sawdust. Amplicon sequencing targeting bacteria, archaea, and fungi was used to explore the microbes present in biofuel production and anaerobic digestion of wastewater and sludge. Biofilm-forming lactic acid bacteria and wild yeasts were identified in fermentation samples of a full-scale plant that uses biowaste as feedstock. During the 20-month monitoring period, the anaerobic digester adapted to the bioethanol process waste with a shift in methanogen profile indicating acclimatization to high concentrations of ammonia. Amplicon sequencing does not specifically target living microbes. The same is true for indirect parameters, such as low pH, metabolites, or genes of lactic acid bacteria. Since rapid identification of living microbes would be indispensable for process management, a commercial method was tested that detects them by measuring the rRNA of selected microbial groups. Small-scale testing indicated that the method gives results comparable with plate counts and microscopic counting, especially for bacterial quantification. The applicability of the method was verified in an industrial bioethanol plant, inspecting the clean-in-place process quality and detecting viability during yeast separation. The results supported it as a fast and promising tool for monitoring microbes throughout industrial bioethanol processes.

Evaluation of Nanofiltration Membranes for the Purification of Monosaccharides: Influence of pH, Temperature, and Sulfates on the Solute Retention and Fouling.

Furfural, acetic acid, and sulfates are found in the hemicellulose (HMC) fraction of lignocellulosic biomass. Separation of furfural, acetic acid, and sulfates from monosaccharides by four nanofiltration (NF) membranes was evaluated with a model solution of glucose, xylose, furfural, acetic acid, and sulfates. Results showed that Alfa Laval NF99HF is the most promising membrane to purify monosaccharides, with the retentions of xylose (85%), glucose (95%), and with the minimum sulfate retention. pH has the highest impact on the retention of all solutes and there is no significant effect of temperature on the retentions of sulphates and acetic acid. Lower pH and temperature are favored to maximize the monosaccharide retention and to remove acetic acid while retaining more furfural with the monosaccharides. Moreover, fouling tendency is maximized at lower pH and higher temperatures. According to the statistical analysis, the retentions of glucose, xylose, furfural, sulfates, and acetic acid are 95%, 90%, 20%, 88%, and 0%, respectively at pH 3 and 25 °C. The presence of sulfates favors the separation of acetic acid and furfural from monosaccharides.

Effective saccharification of lignocellulosic barley straw by mechanocatalytical pretreatment using potassium pyrosulfate as a catalyst.

The catalytic conversion of lignocellulosic biomass is attractive due to the feasible generation of valuable products such as reducing sugars which constitute the basic substrates for chemical and transportation fuel production, as well as the production of renewable hydrogen. This study shows the efficient conversion of lignocellulose, especially hemicellulose, into reducing sugars such as xylose and galactose, by mechanocatalysis using potassium pyrosulfate, K2S2O7, as an effective salt catalyst. Ball milling was performed, introducing a mechanical force which, combined with chemical pretreatment, leads to reducing sugar yields (40%) almost as high as when commonly used sulfuric acid was employed. Kinetic experiments as well as the optimization of the saccharification process are presented.

Solid acid-catalyzed depolymerization of barley straw driven by ball milling.

This study describes a time and energy saving, solvent-free procedure for the conversion of lignocellulosic barley straw into reducing sugars by mechanocatalytical pretreatment. The catalytic conversion efficiency of several solid acids was tested which revealed oxalic acid dihydrate as a potential catalyst with high conversion rate. Samples were mechanically treated by ball milling and subsequently hydrolyzed at different temperatures. The parameters of the mechanical treatment were optimized in order to obtain sufficient amount of total reducing sugar (TRS) which was determined following the DNS assay. Additionally, capillary electrophoresis (CE) and Fourier transform infrared spectrometry (FT-IR) were carried out. Under optimal conditions TRS 42% was released using oxalic acid dihydrate as a catalyst. This study revealed that the acid strength plays an important role in the depolymerization of barley straw and in addition, showed, that the oxalic acid-catalyzed reaction generates low level of the degradation product 5-hydroxymethylfurfural (HMF).