Bioprocess-Technological Potential of Irradiation-Based Fungal Pretreatment Platform Relevant to Lignocellulolytic Biocascade.

Lignocellulose-decaying fungal bioplatforms available are not commercially accessible and are limited to short-term use. In this study, those limitations were overcome by developing a platform using water-soaked rice straw (RS) biodegraded by irradiation-based fungal pretreatment (IBFP). This eco-friendly system increased the ability of RS to biodegrade and ferment without the generation of inhibitory compounds. When processed RS (i.e., with a water-soaking ratio of 81 % and irradiation dose of 80 kGy at 1 MeV and 0.12 mA) was pretreated with Dichomitus squalens for 9 days, the sugar yield was 58.5 % of the theoretical maximum. This sugar yield was comparable to that obtained with unirradiated RS for 15 days, which was 57.9 %. Furthermore, the ethanol concentration of 9.7 g L(-1) provided a yield of 58.1 %; the theoretical maximum and productivity at 0.40 g L(-1) h(-1) were determined after simultaneous saccharification and fermentation for 24 h. In addition, microscopic images revealed that IBFP induced predominant ultrastructural modifications to the surface of cell wall fibers. The peroxidative profiles for different biosystems were analyzed in order to understand substrate-specific biocascades based on the differences in biomass components. The activation level of core lignocellulolysis-related factors was analogous under the optimized conditions of each system.

Bioprocess Evaluation of Water Soaking-Based Microbiological Biodegradation with Exposure of Cellulosic Microfibers Relevant to Bioconversion Efficiency.

To verify the interconnective relationship between biodegradation efficiency and microfibril structure, recalcitrant rice straw (RS) was depolymerized using water soaking-based microbiological biodegradation (WSMB). This eco-friendly biosystem, which does not predominantly generate inhibitory metabolites, could increase both the hydrolytic accessibility and fermentation efficiency of RS. In detail, when swollen RS (with Fenton cascades) was simultaneously bio-treated with Phanerochaete chrysosporium for 12 days, the biodegradability was 65.0 % of the theoretical maximum at the stationary phase. This value was significantly higher than the 30.3 % measured from untreated RS. Similarly, the WSMB platform had an effect on the yield enhancement of ethanol productivity of 32.5 %. However, uniform exposure of fibril polymers appeared to have little impact on bioconversion yields. Additionally, the proteomic pools of the WSMB system were analyzed to understand either substrate-specific or nonspecific biocascades based on the change in microcomposite materials. Remarkably, regardless of modified microfibril chains, the significant pattern of 14 major proteins (|fold| > 2) was reasonably analogous in both systems, especially for lignocellulolysis-related targets.

Effective inactivation of Saccharomyces cerevisiae in minimally processed Makgeolli using low-pressure homogenization-based pasteurization.

In order to address the limitations associated with the inefficient pasteurization platform used to make Makgeolli, such as the presence of turbid colloidal dispersions in suspension, commercially available Makgeolli was minimally processed using a low-pressure homogenization-based pasteurization (LHBP) process. This continuous process demonstrates that promptly reducing the exposure time to excessive heat using either large molecules or insoluble particles can dramatically improve internal quality and decrease irreversible damage. Specifically, optimal homogenization increased concomitantly with physical parameters such as colloidal stability (65.0% of maximum and below 25-µm particles) following two repetitions at 25.0 MPa. However, biochemical parameters such as microbial population, acidity, and the presence of fermentable sugars rarely affected Makgeolli quality. Remarkably, there was a 4.5-log reduction in the number of Saccharomyces cerevisiae target cells at 53.5°C for 70 sec in optimally homogenized Makgeolli. This value was higher than the 37.7% measured from traditionally pasteurized Makgeolli. In contrast to the analytical similarity among homogenized Makgeollis, our objective quality evaluation demonstrated significant differences between pasteurized (or unpasteurized) Makgeolli and LHBP-treated Makgeolli. Low-pressure homogenization-based pasteurization, Makgeolli, minimal processing-preservation, Saccharomyces cerevisiae, suspension stability.

Downstream optimization of fungal-based simultaneous saccharification and fermentation relevant to lignocellulosic ethanol production.

To support the inefficient limitation of long-term biosystem by well-known simultaneous saccharification and fermentation (SSF), electron beam irradiated rice straw (at 80 kGy, 1 MeV, and 0.12 mA) was fermented using fungal-based simultaneous saccharification and fermentation (FBSSF) by saprophytic zygomycetes Mucor indicus. Based on the growth optimization (by response surface methodology), this eco-friendly bioprocess either without metabolic inhibitors (especially furfurals and acetic acids) or byproducts (especially glycerols) significantly increased the biodegradability and fermentability of lignocellulosic rice straw. Specifically, when irradiated straw was simultaneously bioconverted by M. indicus for 48 h, the ethanol yield was 57.2% of the theoretical maximum. This value was on the similar level as the 59.8% (for 144 h) measured from processed straw by well-known SSF. Furthermore, after FBSSF for 144 h based on large-scale mass balance, the ethanol concentration and production yield, and productivity were 34.6 g/L, 72.3% of the theoretical maximum, and 0.24 g/L/h, respectively.

Lignocellulose depolymerization occurs via an environmentally adapted metabolic cascades in the wood-rotting basidiomycete Phanerochaete chrysosporium.

Plant biomass can be utilized by a lignocellulose-degrading fungus, Phanerochaete chrysosporium, but the metabolic and regulatory mechanisms involved are not well understood. A polyomics-based analysis (metabolomics, proteomics, and transcriptomics) of P. chrysosporium has been carried out using statistically optimized conditions for lignocellulolytic reaction. Thirty-nine metabolites and 123 genes (14 encoded proteins) that consistently exhibited altered regulation patterns were identified. These factors were then integrated into a comprehensive map that fully depicts all signaling cascades involved in P. chrysosporium. Despite the diversity of these cascades, they showed complementary interconnection among themselves, ensuring the efficiency of passive biosystem and thereby yielding energy expenditure for the cells. Particularly, many factors related to intracellular regulatory networks showed compensating activity in homeostatic lignocellulolysis. In the main platform of proactive biosystem, although several deconstruction-related targets (e.g., glycoside hydrolase, ureidoglycolate hydrolase, transporters, and peroxidases) were systematically utilized, well-known supporters (e.g., cellobiose dehydrogenase and ferroxidase) were rarely generated.

Extracellular breakdown of lignocellulosic biomass by Dichomitus squalens: peroxidation-based platform and homeostatic regulation.

Dichomitus squalens, a wood-rotting fungus, can utilize recalcitrant biomass for growth; however, the extracellular metabolic processes involved are not well understood. A systematic target analysis of D. squalens has been carried out using optimized conditions for lignocellulolysis. Seven encoded proteins and 58 metabolites that consistently exhibited altered regulation patterns were identified. The predicted regulators were then vertically combined into a comprehensive network that included entire signaling pathways involved in D. squalens. Despite the diversity of these pathways, they showed complementary cooperation among themselves, ensuring the efficiency of active biodegradation and thereby yielding energy saving for the cells. Although several hydrolytic enzymes (e.g., polysaccharide-degrading enzymes, α/ß-hydrolase, and peroxidases) were systematically utilized, peroxidative controllers (e.g., glyoxal oxidase and P450-oxidoreductase) were rarely generated.

Process evaluation of electron beam irradiation-based biodegradation relevant to lignocellulose bioconversion.

In order to solve the inefficient problem of long-term biodegradation by wood-decaying fungus, rice straw (RS) was depolymerized using electron beam irradiation-based biodegradation (EBIBB). This environment-friendly program without the use of inhibitory byproducts significantly increased the digestibility and fermentability of RS. Specifically, when irradiated RS was simultaneously biodegraded by Phanerochaete chrysosporium for 10 days, the sugar yield was 65.5% of the theoretical maximum. This value was on the same level as the 64.8% (for 15 days) measured from unirradiated RS. In case of fermentability, similarly, EBIBB program had an effect on time/energy saving. Furthermore, the transcriptomic profiles under different biosystem were analyzed in order to verify possible substrate-specific regulation based on change of lignocellulosic components. Interestingly, the overall correlation based on the bias (upregulation or downregulation) was reasonably analogous, especially lignocellulolysis-related genes.

Complementary substrate-selectivity of metabolic adaptive convergence in the lignocellulolytic performance by Dichomitus squalens.

The lignocellulolytic platform of the wood-decaying organism Dichomitus squalens is important for production of biodegradable elements; however, the system has not yet been fully characterized. In this study, using statistical target optimization, we analysed substrate selectivity based on a variety of D. squalens metabolic pathways using combined omics tools. As compared with the alkali-lignin (AL) programme, the rice straw (RS) programme has the advantage of multilayered signalling to regulate cellulolytic-related genes or to connect their pathways. The spontaneous instability of the AL programme was accelerated by harsh starvation as compared with that of the RS programme. Therefore, the AL programme converged on cellular maintenance much easier and more rapidly. However, regardless of external substrate/concentration type, the compensatory pattern of the major targets (especially peroxidases and growth regulators) was similar, functioning to maintain cellular homeostasis. Interestingly, ligninolytic-mediated targets under non-kaleidoscopic conditions were induced by a substrate-input-control, and especially this mechanism had an important effect on the early stages of the biodegradation process. This optimized target analysis could be used to understand lignocellulolytic network and to improve downstream efficiency.

Electron beam irradiation enhances the digestibility and fermentation yield of water-soaked lignocellulosic biomass.

In order to overcome the limitation of commercial electron beam irradiation (EBI), lignocellulosic rice straw (RS) was pretreated using water soaking-based electron beam irradiation (WEBI). This environment-friendly pretreatment, without the formation (or release) of inhibitory compounds (especially hydroxymethylfurfural and furfural), significantly increased the enzymatic hydrolysis and fermentation yields of RS. Specifically, when water-soaked RS (solid:liquid ratio of 100%) was treated with WEBI doses of 1 MeV at 80 kGy, 0.12 mA, the glucose yield after 120 h of hydrolysis was 70.4% of the theoretical maximum. This value was predominantly higher than the 29.5% and 52.1% measured from untreated and EBI-treated RS, respectively. Furthermore, after simultaneous saccharification and fermentation for 48 h, the ethanol concentration, production yield, and productivity were 9.3 g/L, 57.0% of the theoretical maximum, and 0.19 g/L h, respectively. Finally, scanning electron microscopy images revealed that WEBI induced significant ultrastructural changes to the surface of lignocellulosic fibers.

Biological pretreatment of rice straw by fermenting with Dichomitus squalens.

Rice straw was fermented by a wood-rot fungus Dichomitus squalens as a biological pretreatment, to increase the enzymatic digestibility of lignocellulose and promote cellulose hydrolysis. Response surface methodology was employed to optimize the fermentation medium of D. squalens for achieving the maximum volumetric activity of manganese peroxidase. The fermentation of rice straw by D. squalens for 15 days resulted in the enzymatic digestibility of 58.1% of theoretical glucose yield for the remaining glucan. In addition, a significant reduction in the crystallinity index and microstructural changes in the fermented rice straw were revealed. When the fungal-fermented rice straw was used as a substrate for ethanol production in simultaneous saccharification and fermentation, the ethanol production yield and productivity were 54.2% of the theoretical maximum and 0.39 g/L/hour, respectively, after 24 hours.

Fungal pretreatment of lignocellulose by Phanerochaete chrysosporium to produce ethanol from rice straw.

Phanerochaete chrysosporium is a wood-rot fungus that is capable of degrading lignin via its lignolytic system. In this study, an environmentally friendly fungal pretreatment process that produces less inhibitory substances than conventional methods was developed using P. chrysosporium and then evaluated by various analytical methods. To maximize the production of manganese peroxidase, which is the primary lignin-degrading enzyme, culture medium was optimized using response surface methodologies including the Plackett-Burman design and the Box-Behnken design. Fermentation of 100 g of rice straw feedstock containing 35.7 g of glucan (mainly in the form of cellulose) by cultivation with P. chrysosporium for 15 days in the media optimized by response surface methodology was resulted in a yield of 29.0 g of glucan that had an enzymatic digestibility of 64.9% of the theoretical maximum glucose yield. In addition, scanning electronic microscopy, confocal laser scanning microscopy, and X-ray diffractometry revealed significant microstructural changes, fungal growth, and a reduction of the crystallinity index in the pretreated rice straw, respectively. When the fungal-pretreated rice straw was used as a substrate for ethanol production in simultaneous saccharification and fermentation (SSF) for 24 h, the ethanol concentration, production yield and the productivity were 9.49 g/L, 58.2% of the theoretical maximum, and 0.40 g/L/h, respectively. Based on these experimental data, if 100 g of rice straw are subjected to fungal pretreatment and SSF, 9.9 g of ethanol can be produced after 96 h, which is 62.7% of the theoretical maximum ethanol yield.

Ethanol production from rice straw using optimized aqueous-ammonia soaking pretreatment and simultaneous saccharification and fermentation processes.

Rice straw was pretreated using aqueous-ammonia solution at moderate temperatures to enable production of the maximum amount of fermentable sugars from enzymatic hydrolysis. The effects of various operating variables including pretreatment temperature, pretreatment time, the concentration of ammonia and the solid-to-liquid ratio on the degree of lignin removal and the enzymatic digestibility were optimized using response surface methodology. The optimal reaction conditions, which resulted in an enzymatic digestibility of 71.1%, were found to be 69 degrees C, 10h and an ammonia concentration of 21% (w/w). The effects of different commercial cellulases and the additional effect of a non-cellulolytic enzyme, xylanase, were also evaluated. Additionally, simultaneous saccharification and fermentation was conducted with rice straw to assess the ethanol production yield and productivity.

Improved enzymatic hydrolysis yield of rice straw using electron beam irradiation pretreatment.

Rice straw was irradiated using an electron beam at currents and then hydrolyzed with cellulase and beta-glucosidase to produce glucose. The pretreatment by electron beam irradiation (EBI) was found to significantly increase the enzyme digestibility of rice straw. Specifically, when rice straw that was pretreated by EBI at 80 kGy at 0.12 mA and 1 MeV was hydrolyzed with 60 FPU of cellulase and 30 CBU of beta-glucosidase, the glucose yield after 132 h of hydrolysis was 52.1% of theoretical maximum. This value was significantly higher than the 22.6% that was obtained when untreated rice straw was used. In addition, SEM analysis of pretreated rice straw revealed that EBI caused apparent damage to the surface of the rice straw. Furthermore, EBI pretreatment was found to increase the crystalline portion of the rice straw. Finally, the crystallinity and enzyme digestibility were found to be strongly correlated between rice straw samples that were pretreated by EBI under different conditions.