Enhanced Delignification of Lignocellulosic Biomass by Recombinant Fungus Phanerochaete chrysosporium Overexpressing Laccases and Peroxidases

2018 ◽  
Vol 28 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Nancy Coconi Linares ◽  
Francisco Fernández ◽  
Achim M. Loske ◽  
Miguel A. Gómez-Lim
Keyword(s):  
Lignocellulosic Biomass ◽  
Phanerochaete Chrysosporium ◽  
Wheat Bran ◽  
Enzyme Production ◽  
Lignin Degradation ◽  
Laccase Activity ◽  
Ligninolytic Enzymes ◽  
Lignocellulose Degradation ◽  
Total Phenolic ◽  
Recombinant Strains

Ligninolytic enzyme production and lignin degradation are typically the rate-limiting steps in the biofuel industry. To improve the efficiency of simultaneous bio-delignification and enzyme production, <i>Phanerochaete chrysosporium</i> was transformed by shock wave-induced acoustic cavitation to co-overexpress 3 peroxidases and 1 laccase and test it on the degradation of sugarcane bagasse and wheat bran. Lignin depolymerization was enhanced by up to 25% in the presence of recombinant fungi in comparison with the wild-type strain. Sugar release on lignocellulose was 2- to 6-fold higher by recombinant fungi as compared with the control. Wheat bran ostensibly stimulated the production of ligninolytic enzymes. The highest peroxidase activity from the recombinant strains was 2.6-fold higher, whereas the increase in laccase activity was 4-fold higher in comparison to the control. The improvement of lignin degradation was directly proportional to the highest peroxidase and laccase activity. Because various phenolic compounds released during lignocellulose degradation have proven to be toxic to cells and to inhibit enzyme activity, a significant reduction (over 40%) of the total phenolic content in the samples treated with recombinant strains was observed. To our knowledge, this is the first report that engineering <i>P. chrysosporium</i> enhances<i></i> biodegradation of lignocellulosic biomass.

scholarly journals Production of Ligninolytic Enzymes by Coptotermes curvignathus Gut Bacteria

2019 ◽  
Vol 23 (1) ◽  
pp. 111-121 ◽  
Author(s):  
Fadilah Ayeronfe ◽  
Angzzas Kassim ◽  
Patricia Hung ◽  
Nadiah Ishak ◽  
Sharfina Syarifah ◽  
...  
Keyword(s):  
Lignocellulosic Biomass ◽  
Lignin Degradation ◽  
Laccase Activity ◽  
Ligninolytic Enzymes ◽  
Maximal Activity ◽  
Kraft Lignin ◽  
Bacterial Strains ◽  
Materials Synthesis ◽  
Culture Supernatants ◽  
Maximum Utilization

Abstract Maximum utilization of lignocellulosic biomass is contingent upon degrading the recalcitrant lignin polymer. Conventional methods employed in delignification require high inputs of energy and chemicals, resulting in the release of highly toxic effluents. The ability of gut flora of Coptotermes curvignathus in lignin degradation was investigated in this study. Production of ligninolytic enzymes was done in an aerated submerged fermentation system with kraft lignin as sole carbon source. The degradation experiment was carried out for 7 days at 30 °C, pH 7. Three potential lignin degraders identified as Bacillus sp., Lysinibacillus sp. and Acinetobacter sp. were successfully isolated. The bacterial growth and secretion of extracellular ligninolytic enzymes confirmed metabolism of kraft lignin by the identified strains. Lysinibacillus sp., a novel lignin degrader showed highest manganese peroxidase (76.36 ± 15.74 U/L) and laccase activity (70.67 ± 16.82 U/L) after 7 and 6 days of incubation respectively, while maximal activity of lignin peroxidase (262.49 ± 0.92 U/L) was recorded after 7 days in culture supernatants of Bacillus sp. With respect to the activity of the secreted enzymes, the lignin degrading potential of these bacterial strains can be explored in the valorisations of lignocellulosic biomass in industrial processes such as pulping, bioethanol production, fine chemicals and materials synthesis.

Enzyme production by immobilized Phanerochaete chrysosporium using airlift reactor

Biologia ◽  
2014 ◽  
Vol 69 (11) ◽  
Author(s):  
Denisse González-Ramírez ◽  
Claudia Muro-Urista ◽  
Ainhoa Arana-Cuenca ◽  
Alejandro Téllez-Jurado ◽  
Aldo González-Becerra
Keyword(s):  
Phanerochaete Chrysosporium ◽  
Peroxidase Activity ◽  
Manganese Peroxidase ◽  
Enzyme Production ◽  
Laccase Activity ◽  
Alginate Beads ◽  
Airlift Reactor ◽  
Airlift Bioreactor ◽  
Brilliant Blue ◽  
Maximum Percentage

AbstractEnzyme production by immobilized Phanerochaete chrysosporium was evaluated in airlift bioreactor and agitated cultures. Free mycelium and immobilized mycelium on alginate beads were tested in the decolourization of 50 and 500 mg/L of Remazol Brilliant Blue R. Dye concentration did not inhibit the fungi development in all tests. In addition, high decolourization percentage of dye was found with free mycelium (99%) in agitated flasks and with immobilized mycelium in airlift (98%). However, decolourization period by immobilized mycelium (120 h) was greater than that by the free mycelium (14 h). Important manganese peroxidase, lignine peroxidase and laccase activities were identified in decolourization process. Manganese peroxidase appeared to be promoted by high dye concentrations during the treatment with immobilized mycelium, but this enzyme was not detected with free mycelium in airlift. Bioreactor prompted also laccase and lignine peroxidase actions in both tests; free mycelium registered a maximum laccase action of 31.569 × 103 U/L in 70 h, whereas immobilized mycelium registered 1.680 × 103 U/L in 170 h, while lignine peroxidase secretion by free P. chrysosporium was higher (1.300 × 103 U/L) than immobilized mycelium (1.250 × 103 U/L). Maximum laccase activity coincided with the maximum percentage of decolourization, however, high peroxidase activity was identified from the start of dye treatment.

scholarly journals Evaluation of the capacity of laccase secretion of four novel isolated white-rot fungal strains in submerged fermentation with lignocellulosic biomass

BioResources ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. 6706-6722
Author(s):  
Qi An ◽  
Wen-Yi Shi ◽  
Yi-Xuan He ◽  
Wen-Yao Hao ◽  
Kai-Yue Ma ◽  
...  
Keyword(s):  
Lignocellulosic Biomass ◽  
Industrial Application ◽  
Submerged Fermentation ◽  
Enzyme Production ◽  
Laccase Activity ◽  
White Rot Fungi ◽  
White Rot ◽  
Pleurotus Pulmonarius ◽  
Toona Sinensis ◽  
Sorghum Straw

The capacity of novel isolated white-rot fungi secreting laccase was evaluated for various kinds of lignocellulosic biomass in submerged fermentation. The laccase secreted by Neofomitella fumosipora Han 386 and Pleurotus pulmonarius Han 527 was significantly faster than that by Coriolopsis trogii Han 751 and Coriolopsis sanguinaria An 282. Maximum laccase from N. fumosipora Han 386 on the four kinds of lignocellulosic biomass tested appeared on the first day. This phenomenon indicated that N. fumosipora Han 386 secreted laccase rapidly compared with other tested strains in this study and showed the superiority in the rate of secreting laccase. Based on the maximum laccase activity, the ability of secreting laccase of C. sanguinaria An 282 was superior to other tested novel isolated strains. On the whole, N. fumosipora Han 386 and P. pulmonarius Han 527 preferred Toona sinensis to produce laccase, C. trogii Han 751 preferred to produce laccase on Populus beijingensis, and C. sanguinaria An 282 grown on Sorghum straw was more suitable for secreting laccase. The results will be helpful for developing bioprocesses using various kinds of lignocellulosic biomass for lignocellulolytic enzyme production and enlarging the number of laccase producing strains for industrial application.

scholarly journals β-Galactosidase-Producing Isolates in Mucoromycota: Screening, Enzyme Production, and Applications for Functional Oligosaccharide Synthesis

2021 ◽  
Vol 7 (3) ◽  
pp. 229
Author(s):  
Bettina Volford ◽  
Mónika Varga ◽  
András Szekeres ◽  
Alexandra Kotogán ◽  
Gábor Nagy ◽  
...  
Keyword(s):  
Wheat Bran ◽  
Enzyme Production ◽  
Specific Activity ◽  
Prebiotic Synthesis ◽  
Crude Enzyme ◽  
Oligosaccharide Synthesis ◽  
Industrial Enzymes ◽  
Chromogenic Agar ◽  
Lichtheimia Ramosa ◽  
Excellent Source

β-Galactosidases of Mucoromycota are rarely studied, although this group of filamentous fungi is an excellent source of many industrial enzymes. In this study, 99 isolates from the genera Lichtheimia, Mortierella, Mucor, Rhizomucor, Rhizopus and Umbelopsis, were screened for their β-galactosidase activity using a chromogenic agar approach. Ten isolates from the best producers were selected, and the activity was further investigated in submerged (SmF) and solid-state (SSF) fermentation systems containing lactose and/or wheat bran substrates as enzyme production inducers. Wheat bran proved to be efficient for the enzyme production under both SmF and SSF conditions, giving maximum specific activity yields from 32 to 12,064 U/mg protein and from 783 to 22,720 U/mg protein, respectively. Oligosaccharide synthesis tests revealed the suitability of crude β-galactosidases from Lichtheimia ramosa Szeged Microbiological Collection (SZMC) 11360 and Rhizomucor pusillus SZMC 11025 to catalyze transgalactosylation reactions. In addition, the crude enzyme extracts had transfructosylation activity, resulting in the formation of fructo-oligosaccharide molecules in a sucrose-containing environment. The maximal oligosaccharide concentration varied between 0.0158 and 2.236 g/L depending on the crude enzyme and the initial material. Some oligosaccharide-enriched mixtures supported the growth of probiotics, indicating the potential of the studied enzyme extracts in future prebiotic synthesis processes.

An integrated paradigm for cellulosic biorefineries: utilization of lignocellulosic biomass as self-sufficient feedstocks for fuel, food precursors and saccharolytic enzyme production

2012 ◽  
Vol 5 (5) ◽  
pp. 7100 ◽  
Author(s):  
Ming W. Lau ◽  
Bryan D. Bals ◽  
Shishir P. S. Chundawat ◽  
Mingjie Jin ◽  
Christa Gunawan ◽  
...  
Keyword(s):  
Lignocellulosic Biomass ◽  
Enzyme Production

scholarly journals Distribution of Phenolic Acids and Antioxidant Activities of Different Bran Fractions from Three Pigmented Wheat Varieties

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Jinli Zhang ◽  
Yan Ding ◽  
Haizhou Dong ◽  
Hanxue Hou ◽  
Xiansheng Zhang
Keyword(s):  
Phenolic Acids ◽  
Wheat Bran ◽  
Phenolic Acid ◽  
Phenolic Content ◽  
Antioxidant Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Total Phenolic ◽  
Scavenging Activity ◽  
Wheat Varieties

Phenolic acid profiles and antioxidant activities of outer bran, coarse bran, and shorts from blue, black, and purple wheat were analyzed. Phenolic acids were mainly in the bound form in pigmented wheat bran fractions. Phenolic acid content decreased in the order of outer bran, coarse bran, and shorts for the three pigmented wheat varieties. HPLC analysis of phenolic extracts demonstrated that the bound form of phenolic acids contained more ferulic, isoferulic, and p-coumaric acids compared to their free counterparts. Among the three pigmented wheat varieties, the bran fractions from blue wheat contained higher bound phenolic acids than the other two pigmented wheat bran fractions, except for purple coarse bran. The blue wheat outer bran had the highest total bound phenolic acid of 3458.71 μg/g while the purple wheat shorts had the lowest of 1730.71 μg/g. The contribution of bound phenolic acids to the total phenolic content and antioxidant activity was significantly higher than that of free phenolic acids. Blue wheat bran fractions had the highest radical scavenging activity against DPPH∙ while those of purple wheat gained the highest ABTS∙+ scavenging activity. High correlations were observed between TPC and radical scavenging capacities for DPPH and ABTS (R2>0.85, P<0.05).

scholarly journals Comparative Evaluation of Agroindustrial Byproducts for the Production of Alkaline Protease by Wild and Mutant Strains ofBacillus subtilisin Submerged and Solid State Fermentation

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Hamid Mukhtar ◽  
Ikramul Haq
Keyword(s):  
Bacillus Subtilis ◽  
Solid State ◽  
Wheat Bran ◽  
Solid State Fermentation ◽  
Soybean Meal ◽  
Alkaline Protease ◽  
Enzyme Production ◽  
Enhanced Production ◽  
Industrial Byproducts ◽  
Agroindustrial Byproducts

The present study describes the screening of different agroindustrial byproducts for enhanced production of alkaline protease by a wild and EMS induced mutant strain ofBacillus subtilisIH-72EMS8. During submerged fermentation, different agro-industrial byproducts were tested which include defatted seed meals of rape, guar, sunflower, gluten, cotton, soybean, and gram. In addition to these meals, rice bran, wheat bran, and wheat flour were also evaluated for protease production. Of all the byproducts tested, soybean meal at a concentration of 20 g/L gave maximum production of the enzyme, that is, 5.74  ±  0.26 U/mL from wild and 11.28  ±  0.45 U/mL from mutant strain, during submerged fermentation. Different mesh sizes (coarse, medium, and fine) of the soybean meal were also evaluated, and a finely ground soybean meal (fine mesh) was found to be the best. In addition to the defatted seed meals, their alkali extracts were also tested for the production of alkaline protease byBacillus subtilis, but these were proved nonsignificant for enhanced production of the enzyme. The production of the enzyme was also studied in solid state fermentation, and different agro-industrial byproducts were also evaluated for enzyme production. Wheat bran partially replaced with guar meal was found as the best substrate for maximum enzyme production under solid state fermentation conditions.

scholarly journals Properties of Thermobifida fusca peroxidase Tfu-1649 and its combined synergistic effects with xylanase on lignocellulose degradation

BioResources ◽  
2020 ◽  
Vol 16 (1) ◽  
pp. 942-953
Author(s):  
Wan-Yu Liao ◽  
Yu-Chun Huang ◽  
Wei-Lin Chen ◽  
Cheng-Yu Chen ◽  
Chao-Hsun Yang
Keyword(s):  
Reducing Sugars ◽  
Plant Biomass ◽  
Synergistic Effects ◽  
Culture Broth ◽  
Lignocellulose Degradation ◽  
Total Phenolic ◽  
Thermobifida Fusca ◽  
Zizania Latifolia ◽  
Thermophilic Actinomycetes ◽  
Biomass Decomposition

Lignocelluloses are comprised of cellulose, hemicellulose, and lignins, which constitute plant biomass. Since peroxidases can degrade lignins, the authors examined peroxidase Tfu-1649, which is secreted from the thermophilic actinomycetes, Thermobifida fusca BCRC 19214. After cultivating for 48 h, the culture broth accumulated 43.66 U/mL of peroxidase activity. The treatment of four types of lignocellulolytic byproducts, i.e., bagasse, corncob, pin sawdust, and Zizania latifolia Turcz husk, with Tfu-1649 alone increased the total phenolic compounds, with limited reducing sugars, but treatment with xylanase, Tfu-11, and peroxidase Tfu-1649 showed synergistic effects. Hence, the co-operative degradation of lignocelluloses by both peroxidase and xylanase could contribute to biomass decomposition and further applications in the agricultural and environmental industries.

scholarly journals Effects of supplementation with phosphorus, calcium and manganese during oil palm frond fermentation by Phanerochaete chrysosporium on ligninase enzyme activity

2020 ◽  
Vol 21 (5) ◽  
Author(s):  
Roni Pazla ◽  
Novirman Jamarun ◽  
Fauzia Agustin ◽  
Mardiati Zain ◽  
Arief Arief ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Phanerochaete Chrysosporium ◽  
Oil Palm ◽  
Manganese Peroxidase ◽  
Crude Protein ◽  
Lignin Degradation ◽  
Lignin Content ◽  
Oil Palm Frond ◽  
Range Test ◽  
Palm Frond

Abstract. Pazla R, Jamarun N, Agustin F, Zain M, Cahyani NO. 2020. Effects of supplementation with phosphorus, calcium and manganese during oil palm frond fermentation by Phanerochaete chrysosporium on ligninase enzyme activity. Biodiversitas 21: 1833-1838. The objective of this study was to evaluate the effects of supplementation with phosphorus (P) in combination with calcium (Ca) and manganese (Mn) during oil palm frond (OPF) fermentation by Phanerochaete chrysosporium on ligninase enzyme activity and lignin degradation. This study was carried out using a randomized complete design with 3 treatments (addition of P, Ca and Mn) and 5 replicates. The following treatments were performed: T1 (P 1000 + Ca 2000 + Mn 150 ppm), T2 (P 1500 + Ca 2000 + Mn 150 ppm), and T3  (P 2000 + Ca 2000 +Mn 150 ppm). The data were subjected to an analysis of variance (ANOVA), and differences between treatment means were tested using Duncan's multiple range test (DMRT). The parameters measured were as follows: lignin peroxidase (LiP) activity (U/mL), manganese peroxidase (MnP) activity (U/mL), crude protein (CP) content (%), crude fiber (CF) content (%) and the decrease in lignin (%). The results revealed a significant increase in LiP activity and CP content and a decrease in the lignin content (p<0.05) by the addition of P in the T3 treatment. However, the treatment nonsignificantly increased (p>0.05) MnP activity and significantly decreased (P<0.05) the CF content. In conclusion, supplementation of the OPF fermentation process with P 2000, Ca 2000, and Mn 150 ppm resulted in the highest ligninase enzyme activity and in decreased lignin content.

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