Microbial electrolysis cells for the production of biohydrogen in dark fermentation - A review.

To assess biohydrogen for future green energy, this review revisited dark fermentation and microbial electrolysis cells (MECs). Hydrogen evolution rate in mesophilic dark fermentation is as high as 192 m3 H2/m3-d, however hydrogen yield is limited. MECs are ideal for improving hydrogen yield from carboxylate accumulated from dark fermentation, whereas hydrogen production rate is too slow in MECs. Hence, improving anode kinetic is very important for realizing MEC biohydrogen. Intracellular electron transfer (IET) and extracellular electron transfer (EET) can limit current density in MECs, which is proportional to hydrogen evolution rate. EET does not limit current density once electrically conductive biofilms are formed on anodes, potentially producing 300 A/m2. Hence, IET kinetics mainly govern current density in MECs. Among parameters associated with IET kinetic, population of anode-respiring bacteria in anode biofilms, biofilm density of active microorganisms, biofilm thickness, and alkalinity are critical for current density.

A processive GH9 family endoglucanase of Bacillus licheniformis and the role of its carbohydrate-binding domain.

One of the critical steps in lignocellulosic deconstruction is the hydrolysis of crystalline cellulose by cellulases. Endoglucanases initially facilitate the breakdown of cellulose in lignocellulosic biomass and are further aided by other cellulases to produce fermentable sugars. Furthermore, if the endoglucanase is processive, it can adsorb to the smooth surface of crystalline cellulose and release soluble sugars during repeated cycles of catalysis before dissociating. Most glycoside hydrolase family 9 (GH9) endoglucanases have catalytic domains linked to a CBM (carbohydrate-binding module) (mostly CBM3) and present the second-largest cellulase family after GH5. GH9 endoglucanases are relatively less characterized. Bacillus licheniformis is a mesophilic soil bacterium containing many glycoside hydrolase (GH) enzymes. We identified an endoglucanase gene, gh9A, encoding the GH9 family enzyme H1AD14 in B. licheniformis and cloned and overexpressed H1AD14 in Escherichia coli. The purified H1AD14 exhibited very high enzymatic activity on endoglucanase substrates, such as ß-glucan, lichenan, Avicel, CMC-Na (sodium carboxymethyl cellulose) and PASC (phosphoric acid swollen cellulose), across a wide pH range. The enzyme is tolerant to 2 M sodium chloride and retains 74% specific activity on CMC after 10 days, the highest amongst the reported GH9 endoglucanases. The full-length H1AD14 is a processive endoglucanase and efficiently saccharified sugarcane bagasse. The deletion of the CBM reduces the catalytic activity and processivity. The results add to the sparse knowledge of GH9 endoglucanases and offer the possibility of characterizing and engineering additional enzymes from B. licheniformis toward developing a cellulase cocktail for improved biomass deconstruction. KEY POINTS: • H1AD14 is a highly active and processive GH9 endoglucanase from B. licheniformis. • H1AD14 is thermostable and has a very long half-life. • H1AD14 showed higher saccharification efficiency than commercial endoglucanase.

Design and evaluation of gas fermentation systems for CO2 reduction to C2 and C4 fatty acids: Non-genetic metabolic regulation with pressure, pH and reaction time.

Addressing the carbon emissions through microbial mediated fermentation is an emerging interest. Custom designed and fabricated gas fermentation (GF) systems were evaluated to optimize the headspace pressure, pH (6.5, 7.5, and 8.5), fermentation time, and substrate concentration by employing enriched homoacetogenic chemolithoautotrophs in non-genetic approach. Headspace pressure showed marked influence on the metabolic conversion of inorganic carbon to acetic and butyric acids with 26% higher productivity than the control (atmospheric pressure). Maximum volatile fatty acid (VFA) yield of 3.7 g/L was observed at alkaline pH (8.5) under 2 bar pressure at carbon load of 10 g/L, 96 h). Acetic (3.0 g/L) and butyric (0.7 g/L) acids were the major products upon conversion of 85% of the inorganic substrate. A better in-situ buffering (ß = 0.048) at pH 8.5 along with higher reductive current (RCC: -4.4 mA) depicted better performance of GF towards CO2 reduction.

Polyhydroxybutyrate production from dark-fermentative effluent and composite grafting with bagasse derived α-cellulose in a biorefinery approach.

The study evaluated the preparation of a biocomposite using waste-derived polyhydroxybutyrate (PHB) and bagasse cellulose (α-cellulose) in a biorefinery approach. PHB was produced using dark fermentation effluent rich in volatile fatty acids (VFA) derived from vegetable waste and α-cellulose was extracted from sugarcane bagasse (SCB). Nutrient limitation induced microbial PHB accumulation, wherein maximum production of 0.28 ± 0.06 g PHB/g DCW (28%) was observed. Confocal examination showed the deposition of PHB granules in the cell cytoplasm and NMR spectrum exhibited a structural correlation. α-Cellulose (0.22 ± 0.02 g α-cellulose/g SCB) was extracted through SCB pretreatment. Thereafter, grafting α-cellulose with PHB offered intermolecular bonding, which resulted in enhanced thermal stability of the biocomposite than corresponding pristine PHB. FE-SEM morphological examination of biocomposite depicted that α-cellulose functioned as a filler to PHB. XRD profiles showed significant decrement in PHB crystallinity, signifying the functional role of α-cellulose as an effective reinforcing agent. Additionally, ether functional group of α-cellulose and ester group of PHB also appeared in XPS analysis of the composite, thus authorizing the effective blending of α-cellulose and PHB. Utilization of bagasse-derived cellulose for strengthening biologically produced PHB expands its applications, while simultaneously addressing the plastic pollution issues. Additional value from this process was further achieved by incorporating the concept of biorefinery, wherein acidogenic fermentation effluents were used for the production of PHA, which enabled the re-entry of products (VFA) to the production cycle, thus achieving circularity.

Hydrothermal liquefaction of biogenic municipal solid waste under reduced H2 atmosphere in biorefinery format.

Municipal solid waste (MSW), an inexorable by-product of anthropogenic activities composes of nearly 50% of the organic (biogenic) fraction. Hydrothermal liquefaction (HTL) was studied to facilitate thermal depolymerization of organic fraction of MSW to biocrude at sub-critical region of water (200 °C; 100 bar pressure) employing H2 induced reducing conditions. Food, vegetable, and composite wastes were evaluated as feedstocks to produce HTL derivatives in the form of liquor (biocrude and aqueous phase), biochar and bio-gas. The biocrude (HTLOF) showed middle oil as major fraction along with C6-C22 compounds. Composite waste resulted in relatively higher yield of biocrude fraction. The aqueous phase (HTLAF) documented the presence of reducing sugars, sotolon and furfurals as major fraction. Biochar (HTLBC) composition showed maximum carbon fraction followed by hydrogen and oxygen. H2 induced reduced condition facilitated conversion of the biogenic MSW at relatively lower input conditions to various biobased fractions cohesively addressing the basic biorefinery requirement.