Optimisation Of Lignin Peroxidase Production From Pycnoporus Sp.

Ligninase or lignin peroxidase is gaining importance for their biotechnology application due to its integral role in the biodegradation of lignin, lignin related aromatic compounds and the potential use in industrial processes such as biopulping, biobleaching and bioremediation. Lignin peroxidase has been extensively studied and has been reported produced by white rot fungus. In this study, a preliminary study was done to screen for the highest lignin peroxidase producer from five locally isolated white rot fungus using agitated and non-agitated culture condition. The highest lignin peroxidase producer, identified as Pycnoporus sp. was selected for the optimisation study. Factorial design approach was chosen to determine the optimum conditions which significantly influence the production of lignin peroxidase by Pycnoporus sp. Optimum condition for the highest lignin peroxidase activity of 51.1 U/L was successfully achieved at 24 mM of nitrogen concentration, agitation speed at 110 rpm, pH 3.5, inoculum concentration of 6 x 106 spores/ml and addition of 1 mM inducer (veratryl alcohol) in carbon limited media

Optimisation of lignin peroxidase production using locally isolated Pycnoporus sp. through factorial design

Lignin peroxidase has been extensively studied and has been reported to produce by white rot fungus. The highest lignin peroxidase producer from local isolates, identified as Pycnoporus sp. was selected for the optimisation study. Factorial design approach was significant to determine the optimum conditions that significantly influenced the production of lignin peroxidase by Pycnoporus sp. Several factors were selected in a range indicated by -1 and +1 for lower and upper level, respectively. The results of ANOVA were analysed to check for the significant factors. Optimum condition for the highest lignin peroxidase activity of 51.1 U L-1 was obtained at 24 mM of nitrogen concentration, agitation speed at 110 rpm, pH 3.5, inoculum concentration of 6x106 spores mL-1 and with the addition of inducer (veratryl alcohol). Considering the results obtained, this statistical design was effective in improving the lignin peroxidase production from Pycnoporus sp

Effect of physical and chemical properties of oil palm empty fruit bunch, decanter cake and sago pith residue on cellulases production by Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2

The effect of cultivation condition of two locally isolated ascomycetes strains namely Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2 were compared in submerged and solid state fermentation. Physical evaluation on water absorption index, solubility index and chemical properties of lignin, hemicellulose and cellulose content as well as the cellulose structure on crystallinity and amorphous region of treated oil palm empty fruit bunch (OPEFB) (resulted in partial removal of lignin), sago pith residues (SPR) and oil palm decanter cake towards cellulases production were determined. Submerged fermentation shows significant cellulases production for both strains in all types of substrates. Crystallinity of cellulose and its chemical composition mainly holocellulose components was found to significantly affect the total cellulase synthesis in submerged fermentation as the higher crystallinity index, and holocellulose composition will increase cellulase production. Treated OPEFB apparently induced the total cellulases from T. asperellum UPM1 and A. fumigatus UPM2 with 0.66 U/mg FPase, 53.79 U/mg CMCase, 0.92 U/mg β-glucosidase and 0.67 U/mg FPase, 47.56 U/mg and 0.14 U/mg β-glucosidase, respectively. Physical properties of water absorption and solubility for OPEFB and SPR also had shown significant correlation on the cellulases production

Production of fermentable sugars using oil palm empty fruit bunch using crude cellulase cocktails from trichoderma asperellum UPM1 and aspergillus fumigatus UPM2 For bioethanol production.

Utilization of oil palm empty fruit bunch (OPEFB) for bioethanol production with crude cellulase cocktails from locally isolated fungi was studied. Enzymatic saccharification of alkaline pretreated OPEFB was done using different cellulase enzyme preparations. Crude cellulase cocktails from Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2 produced 8.37 g/L reducing sugars with 0.17 g/g yield. Production of bioethanol from OPEFB hydrolysate using Baker’s yeast produced approximately 0.59 g/L ethanol, corresponding to 13.8% of the theoretical yield. High reducing sugars concentration in the final fermentation samples resulted from accumulation of non-fermentable sugars such as xylose and cellobiose that were not consumed by the yeast. The results obtained support the possible utilization of OPEFB biomass for bioethanol production in the future

Harnessing the potential of ligninolytic enzymes for lignocellulosic biomass pretreatment

Abundant lignocellulosic biomass from various industries provides a great potential feedstock for the production of value-added products such as biofuel, animal feed, and paper pulping. However, low yield of sugar obtained from lignocellulosic hydrolysate is usually due to the presence of lignin that acts as a protective barrier for cellulose and thus restricts the accessibility of the enzyme to work on the cellulosic component. This review focuses on the significance of biological pretreatment specifically using ligninolytic enzymes as an alternative method apart from the conventional physical and chemical pretreatment. Different modes of biological pretreatment are discussed in this paper which is based on (i) fungal pretreatment where fungi mycelia colonise and directly attack the substrate by releasing ligninolytic enzymes and (ii) enzymatic pretreatment using ligninolytic enzymes to counter the drawbacks of fungal pretreatment. This review also discusses the important factors of biological pretreatment using ligninolytic enzymes such as nature of the lignocellulosic biomass, pH, temperature, presence of mediator, oxygen, and surfactant during the biodelignification process

Lignin pretreatment of oil palm empty fruit bunch using ligninolytic enzyme-mediator and cellulose hydrolysis for fermentable sugar production

Lignocellulosic biomass is the source of cellulosic materials which leads to the fermentable sugars productions. The position of Malaysia as among the top producers and major exporter of palm oil generates abundant of palm oil biomass particularly oil palm empty fruit bunch (OPEFB). Conversion to value added products of such organic acid, compost, bioenergy and enzymes besides sugars which is the key step for most of the processes may overcome the issues in future wastes management.Locally isolated fungus namely Pycnoporus sanguineus was found to be the best ligninolytic enzyme producer among the 20 fungi screened on dyed agar plate.Decolorization of Remazol Brilliant Blue dye added to agar media by the fungi showed the ability to secrete ligninolytic enzyme and further profiling resulted in the production of laccase as the major enzyme followed by manganese peroxidase and lignin peroxidase with the least activities.Physical and chemical structural and compositional particularly lignin acts as a barrier to the enzymatic hydrolysis of cellulose. Appropriate pretreatment to remove lignin is necessary to ensure the access of cellulases enzyme to the cellulosic material embedded in the lignocellulosic matrix. In this study, the application of crude ligninolytic enzyme extracts alone from locally isolated fungi namely Pycnoporus sanguineus to the oil palm empty fruit bunch as a biological pre-treatment was able to produce 19 g/L of fermentable sugars during enzymatic hydrolysis using commercial cellulase and was increased up to approximately 30 g/L with the addition of combination mediator of HBT-Mn (II) and ABTS-Mn (II). Based on Klason lignin determination, the highest lignin removal was achieved at the concentration of 1.5% HBT, 4 mM ABTS and 2 mM manganese ion as much as 8.02%, 8.68% and 3.7%, respectively as compared to raw OPEFB. Klason lignin was also removed by as much as 8.8% at 50°C and 8.16% at 10% of substrate loading. Supported results on the FTIR and GC-MS analysis showed the changes in the structure and chemical bonds of the Cellulase is the key enzyme for the cellulose hydrolysis producing fermentable sugars. The effect of cultivation condition of two locally isolated ascomycetes strains namely Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2 were compared in submerged and solid state fermentation. Physical evaluation on water absorption index, solubility index and chemical properties of lignin, hemicellulose and cellulose content as well as the cellulose structure on crystallinity and amorphous region of treated oil palm empty fruit bunch (OPEFB) (resulted in partial removal of lignin), sago pith residues (SPR) and oil palm decanter cake (OPDC) towards cellulases production were determined. Submerged fermentation shown significant cellulases production for both strains in all types of substrates. Crystallinity of cellulose and its chemical composition mainly holocellulose components was found to significantly affected the total cellulase synthesis in submerged fermentation as the higher crystallinity index and holocellulose composition will increase cellulase production. Treated OPEFB was apparently induced the total cellulases from Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2 with 0.66 U/mg FPase, 53.79 U/mg CMCase, 0.92 U/mg β-glucosidase and 0.67 U/mg FPase, 47.56 U/mg and 0.14 U/mg β-glucosidase,respectively. Physical properties of water absorption and solubility for OPEFB and SPR also had shown significant correlation on the cellulases production.The competency of crude cellulase cocktail from both isolates of Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2 were mixed at 3:2 ratio and applied on the enzymatically treated OPEFB from local isolate were compared with the commercial cellulase and the result obtained was 30% cellulose hydrolysis percentage for ligninolytic-mediator pretreatment and 44% using commercial cellulase. Regardless of the lower individual cellulase from local isolates compared to commercial cellulase.It is therefore suggested that biological approaches alone using ligninolytic enzymemediator as pretreatment and cellulase enzymes produced locally had a promising potential for fermentable sugar productions for OPEFB