Advances in the production of fungi-derived lignocellulolytic enzymes using agricultural wastes

Figures & data

Table 1. Recent applications of various lignocellulolytic enzymes from different wild-type fungal cultures.

Enzyme	Fungal culture	Molecular weight	Optimal temperature and pH for enzyme reaction	Applications	References
Lignin peroxidases	Phanerocheate chrysosporium	46 kDa	-	Biodegradation of toxic synthetic polymer waste	Khatoon et al. 2019
	Paecilomyces sp.	38 kDa, 40 kDa	20–30 °C, pH 2–3	Effluent treatment in pulp and paper industry	Singh et al. 2016
Manganese-dependent peroxidases	Trametes pubescens	55 kDa	40 °C, pH 5	Biodegradation of lignin, decolourisation of textile-dyes	Rekik et al. 2019
	Cerrena unicolour	45 kDa	60 °C, pH 4.5	Decolorization of synthetic dyes, bleaching of denim in textile industry	Zhang et al. 2018
Laccase	Thielavia sp.	70 kDa	70 °C, pH 5	Decolorization of anthraquinone-type dye without redox mediator	Mtibaà et al. 2018
	Albifimbria viridis	66 kDa	25 °C, pH 5	Removal of heavy metal from pollutants, biodegradation of tannin	Ahmadi Khozani et al. 2021
	Penicillium chrysogenum	68 kDa	≥80 °C, pH 5.5	Hydrolytic de-gradation of chemical pollutants	Senthivelan et al. 2019
Endoglucanase	Thermoascus aurantiacus	35 kDa	70 °C, pH 4	Improvement of productivity and nutritive quality in food industry, enhancement of cellulosic garments in textile industry	Dave et al. 2015
	Fomitopsis meliae	-	70 °C, pH 4.8	Refinery and saccharification of biomass in food, animal feed, paper and pulp industries	Patel et al. 2021
	Stachybotrys microspora	55 kDa	50 °C, pH 7	Detergents, textiles and saccharification of lignocelluloses in bio-refinery processes and crop waste management	Ben Hmad et al. 2017
Exoglucanase	Meyerozyma sp.	70 kDa	70 °C, pH 5	Enzymatic hydrolysis of cellulose feedstock for renewable bioethanol production	Kuo et al. 2015
	Aspergillus fumigatus	-	55 °C, pH 4.8	Enzymatic hydrolysis of feedstock to produce organic acids and other chemicals	Mahmood et al. 2013
Glucosidase	Aspergillus unguis	10 kDa	60 °C, pH 6	Enzymatic saccharification of biomass	Rajasree et al. 2013
	Lichtheimia ramosa	-	65 °C, pH 5.5	Production of bioethanol, improvement of food and beverage quality	Garcia et al. 2015
	Penicillium piceum	80 kDa	60 °C, pH 6	Enzymatic hydrolysis of lignocellulosic biomass	Gao et al. 2014
Xylanase	Coprinellus disseminatus	43 kDa	55 °C, pH 6.4	Production in thermo-alkali-tolerant environment, pulp, and paper industry	Agnihotri et al. 2010
	Aspergillus foetidus	29.9 kDa	50 °C, pH 5.3	Production of value-added food ingredient from agro-residues	Chapla et al. 2013
	Myceliophthora heterothallica	27 kDa	65–70 °C, pH 4.5	Pproduction of xylooligosaccharides for use as prebiotics	de Oliveira Simões LC et al. 2019
Mannanase	Aspergillus oryzae	34 kDa	60 °C, pH 5	Bio-bleaching of pulp, augmentation of detergent, upgradation of feed quality, oil drilling	Jana et al. 2018
	Penicillium chrysogenum	30 kDa	50 °C, pH 6	Biobleaching of pulp, production of mannooligosaccharides in food industry	Shalaby et al. 2017
Arabinanase	Aspergillus tubingensis	-	-	Fruit juice processing, wine production	Okado et al. 2020
	Aspergillus thermomutatus	-	-	Deconstruction of lignocellulosic material	Benassi et al. 2014
Galactosidase	Aspergillus niger	76 kDa	50 °C, pH 5	As additive in food processing	Martarello et al. 2019
	Aspergillus terreus	42 kDa	40 °C, pH 6	Development of prebiotics to lactose intolerance patients	Vidya et al. 2020
Glucuronidase	Penicillium purpurogenum	70 kDa	40 °C, pH 6	Highly selective hydrolysis of glycyrrhizin to glycyrrhetinic acid monoglucuronide	Zou et al. 2013
	Chaetomium globosum	-	-	Production of glycyrrhetic acid 3-O-mono-β-D-glucuronide	Gao et al. 2021

− Not mentioned in the study.

Figure 1. Overview of lignocellulolytic enzymes in lignocellulose degradation.

Table 2. Heterologous expressions for lignocellulolytic enzymes and properties of the recombinant enzymes.

Enzyme	Source	Gene	Host organism	Genetic modification	Properties of recombinant enzyme	Reference
Laccase	Pycnoporus sanguineus	pslcc	Aspergillus nidulans	Introduced heterologous gene; overexpressed transcription factor RsmA with aflR promoter	Promoted production (15-fold)	Li et al. 2019
	Coprinopsis cinerea	Lcc9	Pichia pastoris	Introduced heterologous gene	Higher specific activity (3.4-fold), increased stability in pH 4.5–9, improved dye decolourisation	Xu et al. 2019
Lignin peroxidase	Thermothelomyces thermophilus	LiP gene	Aspergillus nidulans	Introduced heterologous gene	Higher enzyme purity, large-scale fed-batch production in stirred-tank bioreactor	Liu et al. 2021
	Phanerochaete chrysosporium	LiPH8	Pichia pastoris	Introduced heterologous gene; optimised condon sequence	Higher activity, enhanced production in 14 L bioreactor, upgraded technical lignin	Majeke et al. 2020
	Phanerochaete chrysosporium	LiPH8	Escherichia coli BL21 (DE3)	Introduced heterologous gene	Easy purification, efficient and effective decolourisation of melanin	Sung et al. 2019
Endoglucanase	Aspergillus fumigatus	DBiNU-1	Kluyveromyces lactis	Introduced heterologous gene	High stability at pH 4–8 and 30–60 °C	Rungrattanakasin et al. 2018
Endoglucanase	Fervidobacterium sp. (77% identity)	Cel776	Saccharomyces cerevisiae	Introduced heterologous gene	Thermophilic, stable in presence of detergent, easy purification with high yield	Escuder-Rodríguez et al. 2022
Glucosidase	Aspergillus niger	bgl	Pichia pastoris	Introduced heterologous gene	High level production (129 IU/mL) in 1 L fermenter	Xia et al. 2018
	Talaromyces amestolkiae	bgl-1	Pichia pastoris	Introduced heterologous gene	High purification yield (>80%), high selectivity	Méndez-Líter et al. 2020
Xylanase	Trichoderma reesei, Neurospora crassa	Xyr1, XLR-1	Penicillium oxalicum	Introduced and mutated heterologous genes	Increased cellulase production (2.8-fold)	Xia et al. 2019
	Thermothelomyces thermophila	ex30a	Pichia pastoris	Introduced heterologous gene	Endo-action similar to bacterial xylanases, exhibited an exo-activity	Katsimpouras et al. 2019
Arabinanase	Thermothielavioides terrestris	Abn93T	Aspergillus nidulans	Introduced heterologous gene	Thermophilic, optimum activity at 70 °C, prolonged stability in pHs 4.0–6.5	Velasco et al. 2020
	Rasamsonia emersonii	Reabn93	Pichia pastoris	Introduced heterologous gene	Thermophilic, maximum activity at 70 °C for 3 h, stable at pH 3–9	An et al. 2022
Glucuronidase	Thermotoga petrophila	Tpgus	Escherichia coli BL21 CodonPlus (DE3)-RIPL	Introduced heterologous gene	Improved thermophilic activity (7-fold), great thermostability at 85 °C for 12 h	Ul and Akram 2019
Endoglucanases, glucosidase, xylanase, xylosidase, acetylxylan esterase	Trichoderma reesei, Aspergillus niger, Aspergillus oryzae, Chrysosporium lucknowense, Talaromyces emersonii, Clostridium cellulovorans	bgl1, xlnD, xlnB, celB, cb2b,cbh, and xynA	Saccharomyces cerevisiae	Introduced and integrated seven heterologous genes; optimised condon sequence	Enabled simultaneous saccharification and fermentation of lignocellulose into ethanol without separate enzyme hydrolysis	Claes et al. 2020

Figure 2. Several production strategies support the fungi-derived lignocellulolytic enzyme production from agriculture wastes and promote enzyme applications.

Khatoon N, Jamal A, Ali MI. 2019. Lignin peroxidase isoenzyme: a novel approach to biodegrade the toxic synthetic polymer waste. Environ Tech. 40(11):1366–1375. doi: 10.1080/09593330.2017.1422550.

 PubMed Web of Science ®Google Scholar

Singh P, Jain P, Verma R, Jagadish RS. 2016. Characterization of lignin peroxidase from Paecilomyces species for decolorisation of pulp and paper mill effluent. J Sci Ind Res. 75(8):500–505.

 Web of Science ®Google Scholar

Rekik H, Zaraî Jaouadi N, Bouacem K, Zenati B, Kourdali S, Badis A, Annane R, Bouanane-Darenfed A, Bejar S, Jaouadi B. 2019. Physical and enzymatic properties of a new manganese peroxidase from the white-rot fungus Trametes pubescens strain i8 for lignin biodegradation and textile-dyes biodecolorization. Int J Biol Macromol. 125:514–525. doi:10.1016/j.ijbiomac.2018.12.053.

 PubMed Web of Science ®Google Scholar

Zhang H, Zhang J, Zhang X, Geng A. 2018. Purification and characterization of a novel manganese peroxidase from white-rot fungus Cerrena unicolor BBP6 and its application in dye decolorization and denim bleaching. Process Biochem. 66:222–229. doi: 10.1016/j.procbio.2017.12.011.

 Web of Science ®Google Scholar

Mtibaà R, Barriuso J, de Eugenio L, Aranda E, Belbahri L, Nasri M, Martínez MJ, Mechichi T. 2018. Purification and characterization of a fungal laccase from the ascomycete Thielavia sp. and its role in the decolorization of a recalcitrant dye. Int J Biol Macromol. 120(PB):1744–1751. doi: 10.1016/j.ijbiomac.2018.09.175.

 PubMedGoogle Scholar

Ahmadi Khozani M, Emtiazi G, Aghaei SS, Ghasemi SM, Zolfaghari MR. 2021. Application of fungal laccase for heavy metals precipitation using tannin as a natural mediator. Int J Environ Sci Te. 18(8):2335–2344. doi: 10.1007/s13762-020-02992-7.

 Web of Science ®Google Scholar

Senthivelan T, Kanagaraj J, Panda RC, Narayani T. 2019. Screening and production of a potential extracellular fungal laccase from Penicillium chrysogenum: Media optimization by response surface methodology (RSM) and central composite rotatable design (CCRD). Biotechnol Reports. 23:e00344. doi: 10.1016/j.btre.2019.e00344.

 Google Scholar

Dave BR, Sudhir AP, Subramanian RB. 2015. Purification and properties of an endoglucanase from Thermoascus aurantiacus. Biotechnol Reports. 6:85–90. doi: 10.1016/j.btre.2014.11.004.

 Google Scholar

Patel A, Divecha J, Shah A. 2021. Fomitopsis meliae CFA 2, a novel brown rot for endoglucanase: emphasis towards enhanced endoglucanase production by statistical approach. Mycology. 12(4):325–340. doi: 10.1080/21501203.2021.1918277.

 PubMed Web of Science ®Google Scholar

Ben Hmad I, Boudabbous M, Belghith H, Gargouri A. 2017. A novel ionic liquid-stable halophilic endoglucanase from Stachybotrys microspora. Process Biochem. 54:59–66. doi: 10.1016/j.procbio.2017.01.007.

 Web of Science ®Google Scholar

Kuo HW, Zeng JK, Wang PH, Chen WC. 2015. A novel exo-glucanase explored from a Meyerozyma sp. fungal strain. Adv Enzyme Res. 3(03):53–65. doi: 10.4236/aer.2015.33006.

 Google Scholar

Mahmood RT, Asad MJ, Mehboob N, Mushtaq M, Gulfraz M, Asgher M, Minhas NM, Hadri SH. 2013. Production, purification, and characterization of exoglucanase by Aspergillus fumigatus. Appl Biochem Biotechnol. 170(4):895–908. doi: 10.1007/s12010-013-0227-x.

 PubMed Web of Science ®Google Scholar

Rajasree KP, Mathew GM, Pandey A, Sukumaran RK. 2013. Highly glucose tolerant β-glucosidase from Aspergillus unguis: NII 08123 for enhanced hydrolysis of biomass. J Ind Microbiol Biotechnol. 40(9):967–975. doi: 10.1007/s10295-013-1291-5.

 PubMed Web of Science ®Google Scholar

Garcia NFL, da Silva Santos FR, Gonçalves FA, da Paz MF, Fonseca GG, Leite RSR. 2015. Production of β-glucosidase on solid-state fermentation by Lichtheimia ramosa in agroindustrial residues: Characterization and catalytic properties of the enzymatic extract. Electron J Biotechn. 18(4):314–319. doi: 10.1016/j.ejbt.2015.05.007.

 Web of Science ®Google Scholar

Gao L, Gao F, Jiang X, Zhang C, Zhang D, Wang L, Wu G, Chen S. 2014. Biochemical characterization of a new β-glucosidase (Cel3E) from Penicillium piceum and its application in boosting lignocelluloses bioconversion and forming disaccharide inducers: New insights into the role of β-glucosidase. Process Biochem. 49(5):768–774. doi: 10.1016/j.procbio.2014.02.012.

 Web of Science ®Google Scholar

Agnihotri S, Dutt D, Tyagi CH, Kumar A, Upadhyaya JS. 2010. Production and biochemical characterization of a novel cellulase-poor alkali-thermo-tolerant xylanase from Coprinellus disseminatus SW-1 NTCC 1165. World J Microbiol Biotechnol. 26(8):1349–1359. doi: 10.1007/s11274-010-0307-9.

 Web of Science ®Google Scholar

Chapla D, Dholakiya S, Madamwar D, Shah A. 2013. Characterization of purified fungal endoxylanase and its application for production of value added food ingredient from agroresidues. Food Bioprod Process. 91(4):682–692. doi: 10.1016/j.fbp.2013.08.005.

 Google Scholar

de Oliveira Simões LC, da Silva RR, de Oliveira Nascimento CE, Boscolo M, Gomes E, da Silva R. 2019. Purification and physicochemical characterization of a novel thermostable xylanase secreted by the fungus Myceliophthora heterothallica F.2.1.4. Appl Biochem Biotechnol. 188(4):991–1008. doi: 10.1007/s12010-019-02973-8.

 PubMed Web of Science ®Google Scholar

Jana UK, Suryawanshi RK, Prajapati BP, Soni H, Kango N. 2018. Production optimization and characterization of mannooligosaccharide generating Β-mannanase from Aspergillus oryzae. Bioresour Technol. 268:308–314. doi:10.1016/j.biortech.2018.07.143.

 PubMed Web of Science ®Google Scholar

Shalaby ASG, Esawy MA, Hussein MDM. 2017. Comparative study between free and immobilized Penicillium chrysogenum mannanase: A local fungal isolate. J Appl Pharm Sci. 7(6):97–104. doi: 10.7324/JAPS.2017.70613.

 Google Scholar

Okado N, Sugi M, Kasamoto S, Mizuhashi F, Roberts A, Danielewska-Nikiel B, Sulaiman C, Pham S. 2020. Safety evaluation of arabinase (arabinan endo-1,5-α-L-arabinanase) from Aspergillus tubingensis. Food Sci Nutr. 8(1):456–478. doi: 10.1002/fsn3.1329.

 PubMed Web of Science ®Google Scholar

Benassi VM, de Lucas RC, Jorge JA, de LT de M PM. 2014. Screening of thermotolerant and thermophilic fungi aiming β-xylosidase and arabinanase production. Braz J Microbiol. 45(4):1459–1467. doi: 10.1590/S1517-83822014000400042.

 PubMed Web of Science ®Google Scholar

Martarello RD, Cunha L, Cardoso SL, de Freitas MM, Silveira D, Fonseca-Bazzo YM, Homem-de-Mello M, Filho EXF, Magalhães PO. 2019. Optimization and partial purification of beta-galactosidase production by Aspergillus niger isolated from Brazilian soils using soybean residue. AMB Express. 9(1):81. doi: 10.1186/s13568-019-0805-6.

 PubMedGoogle Scholar

Vidya B, Palaniswamy M, Angayarkanni J, Ayub Nawaz K, Thandeeswaran M, Krishna Chaithanya K, Tekluu B, Muthusamy K, Gopalakrishnan VK. 2020. Purification and characterization of β-galactosidase from newly isolated Aspergillus terreus (KUBCF1306) and evaluating its efficacy on breast cancer cell line (MCF-7). Bioorg Chem. 94:103442. doi:10.1016/j.bioorg.2019.103442.

 PubMed Web of Science ®Google Scholar

Zou S, Liu G, Kaleem I, Li C. 2013. Purification and characterization of a highly selective glycyrrhizin-hydrolyzing β-glucuronidase from Penicillium purpurogenum Li-3. Process Biochem. 48(2):358–363. doi: 10.1016/j.procbio.2012.12.008.

 Web of Science ®Google Scholar

Gao B, Xiao Y, Zhang Q, Sun J, Zhang Z, Zhu D. 2021. Concurrent production of glycyrrhetic acid 3-O-mono-β-d-glucuronide and lignocellulolytic enzymes by solid-state fermentation of a plant endophytic Chaetomium globosum. Bioresour Bioprocess. 8(1):88. doi: 10.1186/s40643-021-00441-y.

 PubMed Web of Science ®Google Scholar

Li W, Yu J, Li Z, Yin WB. 2019. Rational design for fungal laccase production in the model host Aspergillus nidulans. Sci China Life Sci. 62(1):84–94. doi: 10.1007/s11427-017-9304-8.

 PubMed Web of Science ®Google Scholar

Xu G, Wang J, Yin Q, Fang W, Xiao Y, Fang Z. 2019. Expression of a thermo- and alkali-philic fungal laccase in Pichia pastoris and its application. Protein Expr Purif. 154:16–24. doi:10.1016/j.pep.2018.09.015.

 PubMed Web of Science ®Google Scholar

Liu E, Segato F, Wilkins MR. 2021. Fed-batch production of Thermothelomyces thermophilus lignin peroxidase using a recombinant Aspergillus nidulans strain in stirred-tank bioreactor. Bioresour Technol. 325:124700. doi:10.1016/j.biortech.2021.124700.

 PubMed Web of Science ®Google Scholar

Majeke BM, García-Aparicio M, Biko OD, Viljoen-Bloom M, van Zyl WH, Görgens JF. 2020. Synergistic codon optimization and bioreactor cultivation toward enhanced secretion of fungal lignin peroxidase in Pichia pastoris: Enzymatic valorization of technical (industrial) lignins. Enzyme Microb Technol. 139:109593. doi: 10.1016/j.enzmictec.2020.109593.

 PubMed Web of Science ®Google Scholar

Sung HJ, Khan MF, Kim YH. 2019. Recombinant lignin peroxidase-catalyzed decolorization of melanin using in-situ generated H2O2 for application in whitening cosmetics. Int J Biol Macromol. 136:20–26. doi:10.1016/j.ijbiomac.2019.06.026.

 PubMed Web of Science ®Google Scholar

Rungrattanakasin B, Premjet S, Thanonkeo S, Klanrit P, Thanonkeo P. 2018. Cloning and expression of an endoglucanase gene from the thermotolerant fungus Aspergillus fumigatus DBiNU-1 in Kluyveromyces lactis. Braz J Microbiol. 49(3):647–655. doi: 10.1016/j.bjm.2017.10.001.

 PubMed Web of Science ®Google Scholar

Escuder-Rodríguez JJ, González-Suarez M, deCastro ME, Saavedra-Bouza A, Becerra M, González-Siso MI. 2022. Characterization of a novel thermophilic metagenomic GH5 endoglucanase heterologously expressed in Escherichia coli and Saccharomyces cerevisiae. Biotechnol Biofuels. 15(1):76. doi: 10.1186/s13068-022-02172-4.

 Web of Science ®Google Scholar

Xia Y, Yang L, Xia L. 2018. High-level production of a fungal β-glucosidase with application potentials in the cost-effective production of Trichoderma reesei cellulase. Process Biochem. 70:55–60. doi: 10.1016/j.procbio.2018.03.031.

 Web of Science ®Google Scholar

Méndez-Líter JA, Nieto-Domínguez M, Fernández De Toro B, González Santana A, Prieto A, Asensio JL, Cañada FJ, De Eugenio LI, Martínez MJ. 2020. A glucotolerant β-glucosidase from the fungus Talaromyces amestolkiae and its conversion into a glycosynthase for glycosylation of phenolic compounds. Microb Cell Fact. 19(1):127. doi: 10.1186/s12934-020-01386-1.

 PubMed Web of Science ®Google Scholar

Xia C, Li Z, Xu Y, Yang P, Gao L, Yan Q, Li S, Wang Y, Qu Y, Song X. 2019. Introduction of heterologous transcription factors and their target genes into Penicillium oxalicum leads to increased lignocellulolytic enzyme production. Appl Microbiol Biotechnol. 103(6):2675–2687. doi: 10.1007/s00253-018-09612-y.

 PubMed Web of Science ®Google Scholar

Katsimpouras C, Dedes G, Thomaidis NS, Topakas E. 2019. A novel fungal GH30 xylanase with xylobiohydrolase auxiliary activity. Biotechnol Biofuels. 12(1):3619–3658. doi: 10.1186/s13068-019-1455-2.

 Google Scholar

Velasco J, Oliva B, Gonçalves AL, Lima AS, Ferreira G, França BA, Mulinari EJ, Gonçalves TA, Squina FM, Kadowaki MAS, et al. 2020. Functional characterization of a novel thermophilic exo-arabinanase from Thermothielavioides terrestris. Appl Microbiol Biotechnol. 104(19):8309–8326. doi: 10.1007/s00253-020-10806-6.

 PubMed Web of Science ®Google Scholar

An J, Xu W, Meng X, Chen G, Zhang W, Liu W. 2022. Biochemical characterization of a thermophilic exo-arabinanase from the filamentous fungus Rasamsonia emersonii. J Biosci Bioeng. 133(4):316–322. doi: 10.1016/j.jbiosc.2021.12.010.

 PubMed Web of Science ®Google Scholar

Ul HI, Akram F. 2019. Enhanced production, overexpression and characterization of a hyperthermophilic multimodular GH family 2 β glucuronidase (TpGUS) cloned from Thermotoga petrophila RKU-1T in a mesophilic host. Int J Biol Macromol. 123:1132–1142. doi:10.1016/j.ijbiomac.2018.11.189.

 PubMed Web of Science ®Google Scholar

Claes A, Deparis Q, Foulquié-Moreno MR, Thevelein JM. 2020. Simultaneous secretion of seven lignocellulolytic enzymes by an industrial second-generation yeast strain enables efficient ethanol production from multiple polymeric substrates. Metab Eng. 59:131–141. doi: 10.1016/j.ymben.2020.02.004.

 PubMed Web of Science ®Google Scholar