Development of novel recombinant peroxidase secretion system from Pseudomonas putida for lignin valorisation.

Pseudomonas putida is a promising strain for lignin valorisation. However, there is a dearth of stable and efficient systems for secreting enzymes to enhance the process. Therefore, a novel secretion system for recombinant lignin-depolymerising peroxidase was developed. By adopting a flagellar type III secretion system, P. putida KT-M2, a secretory host strain, was constructed and an optimal secretion signal fusion partner was identified. Application of the dye-decolourising peroxidase of P. putida to this system resulted in efficient oxidation activity of the cell-free supernatant against various chemicals, including lignin model compounds. This peroxidase-secreting strain was examined to confirm its lignin utilisation capability, resulting in the efficient assimilation of various lignin substrates with 2.6-fold higher growth than that of the wild-type strain after 72 h of cultivation. Finally, this novel system will lead efficient bacterial lignin breakdown and utilization through enzyme secretion, paving the way for sustainable lignin-consolidated bioprocessing.

Effect of torrefied biomass on hydrophobicity and mechanical properties of polylactic acid composite.

In this study, the physicochemical properties of torrefied biomass (larch and yellow poplar) were investigated based on torrefaction temperature. The effect of torrefied biomass on the hydrophobicity and mechanical properties of a polylactic acid (PLA) composites was evaluated. Hemicellulose was removed from the biomass during torrefaction, whereas the cellulose and lignin contents increased slightly. The color of the biomass changed from brown to black. The grindability of the torrefied biomass improved as the torrefaction temperature increased, which contributed to the production of fine particles (>100 mesh). A PLA composite was prepared using torrefied biomass (10 %) and polylactic acid. At 280 °C, water contact angle was the highest, regardless of the particle size and biomass species. Tensile strength of the PLA composite was slightly lower than that of PLA alone, regardless of the particle size of torrefied biomass. Nevertheless, the strength increased with the torrefaction temperature, except for larch with a relatively large particle size (<100 mesh). The tensile strength of the control was 68.0 MPa, whereas that of the torrefied biomass ranged from 61.1 to 65.8 MPa.

Efficient ethanol production from dried oil palm trunk treated by hydrothermolysis and subsequent enzymatic hydrolysis.

BACKGROUND: Oil palm trunk (OPT) is a valuable bioresource for the biorefinery industry producing biofuels and biochemicals. It has the distinct feature of containing a large amount of starch, which, unlike cellulose, can be easily solubilized by water when heated and hydrolyzed to glucose by amylolytic enzymes without pretreatment for breaking down the biomass recalcitrance. Therefore, it is suggested as beneficial to extract most of the starch from OPT through autoclaving and subsequent amylolytic hydrolysis prior to pretreatment. However, this treatment requires high capital and operational costs, and there could be a high probability of microbial contamination during starch processing. In terms of biochemical conversion of OPT, this study aimed to develop a simple and efficient ethanol conversion process without any chemical use such as acids and bases or detoxification. RESULTS: For comparison with the proposed efficient ethanol conversion process, OPT was subjected to hydrothermal treatment at 180 °C for 30 min. After enzymatic hydrolysis of PWS, 43.5 g of glucose per 100 g dry biomass was obtained, which corresponds to 81.3 % of the theoretical glucose yield. Through subsequent alcohol fermentation, 81.4 % ethanol yield of the theoretical ethanol yield was achieved. To conduct the proposed new process, starch in OPT was converted to ethanol through enzymatic hydrolysis and subsequent fermentation prior to hydrothermal treatment, and the resulting slurry was subjected to identical processes that were applied to control. Consequently, a high-glucose yield of 96.3 % was achieved, and the resulting ethanol yield was 93.5 %. CONCLUSIONS: The proposed new process was a simple method for minimizing the loss of starch during biochemical conversion and maximizing ethanol production as well as fermentable sugars from OPT. In addition, this methodology offers the advantage of reducing operational and capital costs due to minimizing the process for ethanol production by excluding expensive processes related to detoxification prior to enzymatic hydrolysis and fermentation such as washing/conditioning and solid-liquid separation of pretreated slurry. The potential future use of xylose-digestible microorganisms could further increase the ethanol yield from the proposed process, thereby increasing its effectiveness for the conversion of OPT into biofuels and biochemicals.

Sugar yields from sunflower stalks treated by hydrothermolysis and subsequent enzymatic hydrolysis.

To produce fermentable sugar with fewer microbial inhibitors via few processes, batch-type hydrothermal treatments of sunflower stalks were performed, followed by enzymatic hydrolysis of pretreatment slurries with Cellic CTec2 and Cellic HTec2 (9:1, v/v, 0.1 ml/g dry biomass, 8.1FPU). The yields of hemicellulosic sugars were maximized at the pretreatment condition of 180°C for 30 min, while the furfural and 5-hydroxymethyl-2-furaldehyde (HMF) concentrations remained low. The glucose yield, however, was only 67.0% of the theoretical glucose yield (TGY). Either the treatment of raw biomass prior to pretreatment or the post-treatment of pretreated residue prior to enzymatic hydrolysis increased the glucose yield as follows: washing the pretreated solid with solvents (90% TGY)>partial removal of liquid fraction from the pretreatment slurry (PS, 83%)>removal of hot-water extractives from the biomass prior to pretreatment (77%)>prewetting of the biomass (70%).