Cellulolytic Enzyme Activities Research Articles

Improvement of wheat straw hydrolysis by cellulolytic blends of two Penicillium spp.

Co-culture of fungal strains Penicillium janthinellum EMS-UV-8 (E), Penicillium funiculosum strain P (P) and Aspergillus sp. strain G (G) and blending of their crude cellulase were evaluated for improvements in cellulase activities as well as for enhanced hydrolysis of dilute acid pretreated wheat straw (PWS). The blending of crude enzymes of P and E enhanced the hydrolysis of PWS more effectively due to synergism in cellulolytic enzyme activities. Here, three types of blends were made on the basis of equal FPUs, equal protein content or fixed volume containing different proportions of individual enzymes, the former blend hydrolyzed 42.6% of PWS due to the 98%,62%, 64% and 34% synergistic enhancement in endo-glucanase, cellulase (FPU), β-glucosidase and xylanase activities, respectively. Hydrolysis at 10% solid loading of PWS in roller bottle reactor with this blend further enhanced hydrolysis yield to 74% within 24 h, which was much better than the corresponding hydrolysis yields of individual (38.1% by E and 61.5% by P) or the commercial enzyme (62.3%). This study proved that synergistic blends of cellulases from two Penicillium spp. are cost-effective tools for efficient wheat straw hydrolysis for on-site biofuel production.

Addition of metal ions to a (hemi)cellulolytic enzymatic cocktail produced in-house improves its activity, thermostability, and efficiency in the saccharification of pretreated sugarcane bagasse

High activity and stability are essential for (hemi)cellulolytic enzymes used in biomass conversion, while non-productive binding of cellulases to lignin reduces saccharification efficiency and needs to be avoided. One potential strategy is the addition of inexpensive metal ions. This paper describes the influence of divalent metal ions on the activity, thermostability, and saccharification efficiency of (hemi)cellulolytic enzymes produced in-house by Aspergillus niger under solid-state fermentation (SSF). The use of Mn(2+) provided the best (hemi)cellulolytic activity and stability, with an increase in endoglucanase activity of up to 57%. The use of Mn(2+) was then investigated in the saccharification of sugarcane bagasse submitted to acid, steam-explosion, and hydrothermal pretreatments. The addition of Mn(2+) ions at 10mM in the saccharification of acid-pretreated bagasse resulted in a 34% increase in glucose release. These positive effects appeared to be due to a reduction in non-productive enzyme adsorption. The findings suggest that the addition of inexpensive metal ions can help to improve activity, thermostability, and saccharification efficiency of (hemi)cellulolytic enzymes.

Halo-tolerance of Marine-derived Fungi and their Enzymatic Properties

Marine environments are unique habitats for microorganisms and represent uninvestigated areas that possess valuable resources. Fungi may also be important because they contribute to marine ecosystems as decomposers. The aims of this study were to investigate the halo-tolerance of marine-derived fungi and their enzymatic properties. Eighteen fungal strains representing 11 different species were used, which included 17 ascomycetes and a zygomycete. The majority were not affected by salinity and showed endo-glucanase (EG) and β-glucosidase (BGL) activities. Interestingly, the cellulolytic enzyme activity derived from Penicillium chrysogenum FU42 increased with salinity. To investigate whether this increase was due to an adaptation or an innate ability of the species, P. chrysogenum KCTC6933, which originated from a terrestrial environment, was used as a control, and its enzymatic properties were compared. Consequently, P. chrysogenum FU42, which was derived from the ocean, showed unique enzymatic properties that might enable the fungus to live in the ocean and contribute to the nutrient cycle in marine ecosystems.

Investigation of Marine-Derived Fungal Diversity and Their Exploitable Biological Activities.

Marine fungi are potential producers of bioactive compounds that may have pharmacological and medicinal applications. Fungi were cultured from marine brown algae and identified using multiple target genes to confirm phylogenetic placement. These target genes included the internal transcribed spacer (ITS), the nuclear large subunit (LSU), and the β-tubulin region. Various biological activities of marine-derived fungi were evaluated, including their antifungal, antioxidant and cellulolytic enzyme activities. As a result, a total of 50 fungi was isolated from the brown algae Sargassum sp. Among the 50 isolated fungi, Corollospora angusta was the dominant species in this study. The genus Arthrinium showed a relatively strong antifungal activity to all of the target plant pathogenic fungi. In particular, Arthrinium saccharicola KUC21221 showed high radical scavenging activity and the highest activities in terms of filter paper units (0.39 U/mL), endoglucanase activity (0.38 U/mL), and β-glucosidase activity (1.04 U/mL).

Production and assay of cellulolytic enzyme activity of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya abbatoir, Indonesia.

Aim:This study aims to produce and assay cellulolytic enzyme activity (endo-(1,4)-β-D-glucanase, exo-(1,4)-β-D-glucanase, and β-glucosidase, at optimum temperature and optimum pH) of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya Abbatoir, Indonesia.Materials and Methods:To produce enzyme from a single colony of E. cloacae WPL 214, 98 × 1010 CFU/ml of isolates was put into 20 ml of liquid medium and incubated in a shaker incubator for 16 h at 35°C in accordance with growth time and optimum temperature of E. cloacae WPL 214. Further on, culture was centrifuged at 6000 rpm at 4°C for 15 min. Pellet was discarded while supernatant containing cellulose enzyme activity was withdrawn to assay endo-(1,4)-β-D-glucanase, exo-(1,4)-β-D-glucanase, and β-glucosidase.Results:Cellulase enzyme of E. cloacae WPL 214 isolates had endoglucanase activity of 0.09 U/ml, exoglucanase of 0.13 U/ml, and cellobiase of 0.10 U/ml at optimum temperature 35°C and optimum pH 5.Conclusion:E. cloacae WPL 214 isolated from bovine rumen fluid waste produced cellulose enzyme with activity as cellulolytic enzyme of endo-(1,4)-β-D-glucanase, exo-(1,4)-β-D-glucanase and β-glucosidase.

Biomass estimation during macro-scale solid-state fermentation of basidiomycetes using established and novel approaches.

Solid-state fermentation (SSF) has been utilised in food production for millennia and is well suited for the cultivation of basidiomycetes, due to the robustness of the process and the possibility of using lignocellulose as the substrate. Basidiomycetes produce diverse enzymes and various primary and secondary metabolites, many of which have biotechnological potential. The quantification of the fungal biomass present is essential for the characterisation of growth kinetics in processes such as SSF. In SSF, fungi grow into the substrate and use it as a nutrient source. Therefore, direct biomass determination is not possible and indirect methods have to be employed. In the presented study, we compared 11 methods for quantifying fungal biomass during SSF of the basidiomycete Trametes hirsuta in a newly developed laboratory reactor (working volume 10L). The methods were based on measuring the levels of six cell-specific components (ergosterol, glucosamine, nucleic acids, number of fungal nuclei, protein and genomic DNA) and estimations of biological activity (respiration, activities of lignolytic and cellulolytic enzymes, and the glucose and protein contents of the liquid). The methods were evaluated with regards to reproducibility and plausibility of the results, time and resource requirements, possible influential factors, and matrix effects. The most reliable biomass estimates were obtained from measurements of ergosterol content, number of nuclei, and respiration. Thus, these three methods were deemed most suitable for process control and modelling.

Redox-initiated hydrogel system for detection and real-time imaging of cellulolytic enzyme activity.

Invited for this month′s cover is the group of Michael Nash at the University of Munich. The image shows cellulase enzymes digesting cellulose fibers and converting the signal into fluorescent hydrogel, as described in their article. The Full Paper itself is available at 10.1002/cssc.201402428

Redox-initiated hydrogel system for detection and real-time imaging of cellulolytic enzyme activity.

Understanding the process of biomass degradation by cellulolytic enzymes is of urgent importance for biofuel and chemical production. Optimizing pretreatment conditions and improving enzyme formulations both require assays to quantify saccharification products on solid substrates. Typically, such assays are performed using freely diffusing fluorophores or dyes that measure reducing polysaccharide chain ends. These methods have thus far not allowed spatial localization of hydrolysis activity to specific substrate locations with identifiable morphological features. Here we describe a hydrogel reagent signaling (HyReS) system that amplifies saccharification products and initiates crosslinking of a hydrogel that localizes to locations of cellulose hydrolysis, allowing for imaging of the degradation process in real time. Optical detection of the gel in a rapid parallel format on synthetic and natural pretreated solid substrates was used to quantify activity of T. emersonii and T. reesei enzyme cocktails. When combined with total internal reflection fluorescence microscopy and AFM imaging, the reagent system provided a means to visualize enzyme activity in real-time with high spatial resolution (<2 μm). These results demonstrate the versatility of the HyReS system in detecting cellulolytic enzyme activity and suggest new opportunities in real-time chemical imaging of biomass depolymerization.

Digestive tube contents of blood cockle (Anadara granosa) in a tropical mangrove estuary in Malaysia

This study was carried out to clarify the feeding biology of the blood cockle ( Anadara granosa ). We collected blood cockles from 8 stations in the Matang mangrove estuary of Malaysia in July and August 2010. The digestive tube contents of the specimens were stained with Congo red and observed under a light microscope. The results showed blood cockles take in particles containing cellulose as well as phytoplankton such as diatoms. As blood cockles in estuaries are known to exhibit cellulolytic enzyme activity in their digestive gland, the present results indicate blood cockles in estuaries feed on litter supplied from mangrove forests and terrestrial plants.

Establishment of a novel, eco-friendly transgenic pig model using porcine pancreatic amylase promoter-driven fungal cellulase transgenes.

Competition between humans and livestock for cereal and legume grains makes it challenging to provide economical feeds to livestock animals. Recent increases in corn and soybean prices have had a significant impact on the cost of feed for pig producers. The utilization of byproducts and alternative ingredients in pig diets has the potential to reduce feed costs. Moreover, unlike ruminants, pigs have limited ability to utilize diets with high fiber content because they lack endogenous enzymes capable of breaking down nonstarch polysaccharides into simple sugars. Here, we investigated the feasibility of a transgenic strategy in which expression of the fungal cellulase transgene was driven by the porcine pancreatic amylase promoter in pigs. A 2,488bp 5'-flanking region of the porcine pancreatic amylase gene was cloned by the genomic walking technique, and its structural features were characterized. Using GFP as a reporter, we found that this region contained promoter activity and had the potential to control heterologous gene expression. Transgenic pigs were generated by pronuclear microinjection. Founders and offspring were identified by PCR and Southern blot analyses. Cellulase mRNA and protein showed tissue-specific expression in the pancreas of F1 generation pigs. Cellulolytic enzyme activity was also identified in the pancreas of transgenic pigs. These results demonstrated the establishment of a tissue-specific promoter of the porcine pancreatic amylase gene. Transgenic pigs expressing exogenous cellulase may represent a way to increase the intake of low-cost, fiber-rich feeds.

Enzymatic hydrolysis of beech wood lignocellulose at high solid contents and its utilization as substrate for the production of biobutanol and dicarboxylic acids

The development of a cost-effective hydrolysis for crude cellulose is an essential part of biorefinery developments. To establish such high solid hydrolysis, a new solid state reactor with static mixing is used. However, concentrations >10% (w/w) cause a rate and yield reduction of enzymatic hydrolysis. By optimizing the synergetic activity of cellulolytic enzymes at solid concentrations of 9%, 17% and 23% (w/w) of crude Organosolv cellulose, glucose concentrations of 57, 113 and 152gL−1 are reached. However, the glucose yield decreases from 0.81 to 0.72gg−1 at 17% (w/w). Optimal conditions for hydrolysis scale-up under minimal enzyme addition are identified. As result, at 23% (w/w) crude cellulose the glucose yield increases from 0.29 to 0.49gg−1. As proof of its applicability, biobutanol, succinic and itaconic acid are produced with the crude hydrolysate. The potential of the substrate is proven e.g. by a high butanol yield of 0.33gg−1.

Role of microbiocenosis of the gastrointestinal tract in the nutrition of grouse

In order to assess the role of the microbiocenosis of the digestive tract of herbivorous birds in transforming poor forage, the activity of cellulolytic enzymes in all departments of the gastrointestinal tract of the black grouse (Lyrurus tetrix) and Western capercaillie (Tetrao urogallus) was studied. The functional diversity of microbial communities of different departments was also investigated. In both species of birds, nitrogenase and cellobiohydrolase activities were discovered in the digestive tract, with the maximum observed in the cecum.

Bioconversion of lignocellulosic palm byproducts into enzymes and lipid by newly isolated oleaginous fungi

Direct conversion of palm pressed fiber (PPF) and palm empty fruit bunches (EFB) into enzymes and lipid by oleaginous fungi were performed through solid-state fermentation (SSF). Among the strains tested, TSIP9 converted PPF and EFB into lipid with the highest yield of 31.1±1.7mg/gram dry substrate (gds) and 37.5±2.2mg/gds, respectively. It also produced high activity of cellulolytic enzymes. It was identified as Aspergillus tubingensis. The similar fatty acids of its lipid to those of plant oil indicate its suitable use as biodiesel feedstock. The cellulase and xylanase production by this strain was improved when EFB was pretreated with alkaline. When alkaline-pretreated EFB was added with palm kernel cake (PK) as an alternative nitrogen source and the culture conditions were optimized through response surface methodology (RSM), the production of lipid, cellulase and xylanase were increased up to 88.5±4.9mg/gds, 26.1±0.1U/gds and 59.3±0.3U/gds, respectively. This study reveals the potential use of in situ cellulolytic enzymes producing fungi and the optimal conditions for direct conversion of lignocellulosic biomass into lipid.

Symbiotic Behavior during Co-culturing of Clostridium thermocellum NKP-2 and Thermoanaerobacterium thermosaccharolyticum NOI-1 on Corn Hull

The symbiosis of co-culturing between Clostridium thermocellum NKP-2 and Thermoanaerobacterium thermosaccharolyticum NOI-1 is described. An efficient biomass-degrading enriched culture was isolated from soil that contained two different bacterial strains showing homology to C. thermocellum and T. thermosaccharolyticum. The enzymatic system produced from the isolated strains when cultivated individually on corn hulls demonstrated different cellulolytic and xylanolytic enzyme activities. Strain NKP-2 produced cellulose- and xylan-main chain cleaving enzymes such as carboxymethylcellulase (CMCase), avicelase, and xylanase as major enzymes, whereas strain NOI-1 produced primarily short- and side-chain cleaving enzymes such as cellobiohydrolase, β-glucosidase, β-xylosidase, acetyl esterase, and especially α-L-arabinofuranosidase. Enhancement of corn hull utilization, cell growth, and fermentation products (ethanol, butanol, acetic acid, butyric acid, H2, and CO2) was greatly increased during co-culturing compared with individual cultivation of both strains. The symbiotic behavior between both strains was one of mutualism, in which the synergistic degradation of corn hulls by co-action of cellulolytic and xylanolytic enzymes promoted hydrolysis of biomass for growth and fermented products.

Microbial interactions affect sources of priming induced by cellulose

The recently developed 3-source-partitioning approach: addition of 14C labeled organics to soil after C3–C4 vegetation changes, was used to distinguish C sources in three compartments, namely CO2, microbial biomass and dissolved organic C (DOC) during decomposition of labeled cellulose. Microbial community structure (based on PLFA composition) and functions (based on enzyme activities and on microbial growth parameters) revealed mechanisms and drivers of priming effects (PE) induced by cellulose addition.14C-cellulose input caused negative PE within the first week and was accompanied by fast consumption of unlabelled DOC and its incorporation into microbial biomass. Microbial activation however, was not confirmed by substrate-induced respiration, nor by hydrolytic enzymes activity or by PLFA changes. A remarkable exception was a 2-fold increase in protozoan PLFA. Such an increase indicates that microorganisms feeding on cellulose and on DOC were quickly grazed by protozoans acting as a driver of microbial succession. This experimentally demonstrates the functioning of the microbial interactions: protozoan grazers provided for rapid recycling of nutrients and facilitated the succession of cellulose-degrading microorganisms during the second week of cellulose decomposition. An increase in the activity of cellulolytic enzymes caused short-term real PE accompanied by increase in abundance of slow-growing fungi and G(−) bacteria. Long-term real PE observed between 14 and 60 days after cellulose input was due to decomposition of SOM-originated hemicelluloses by fungi and G(+) bacteria. The CO2 released by primed soil organic matter (SOM) decomposition was originated mainly from C younger than 12 years (63%) and only 37% were older than 12 years despite the recent and old C contributed almost equally (51 and 49%, respectively) to SOM under Miscanthus giganteus. This indicates that the SOM pools are involved in PE according to their availability. Despite 71% of the applied cellulose-C was sequestered in the soil, the net soil C-gain amounted only for 28% of the applied cellulose-C after factoring in the C losses by the PE. Our study emphasizes the role of food webs in the PE dynamics: cellulose input served as a driver activating the food chain through the microbial loop.

Transcriptional analysis of selected cellulose-acting enzymes encoding genes of the white-rot fungus Dichomitus squalens on spruce wood and microcrystalline cellulose

The recent discovery of oxidative cellulose degradation enhancing enzymes has considerably changed the traditional concept of hydrolytic cellulose degradation. The relative expression levels of ten cellulose-acting enzyme encoding genes of the white-rot fungus Dichomitus squalens were studied on solid-state spruce wood and in microcrystalline Avicel cellulose cultures. From the cellobiohydrolase encoding genes, cel7c was detected at the highest level and showed constitutive expression whereas variable transcript levels were detected for cel7a, cel7b and cel6 in the course of four-week spruce cultivation. The cellulolytic enzyme activities detected in the liquid cultures were consistent with the transcript levels. Interestingly, the selected lytic polysaccharide monooxygenase (LPMO) encoding genes were expressed in both cultures, but showed different transcription patterns on wood compared to those in submerged microcrystalline cellulose cultures. On spruce wood, higher transcript levels were detected for the lpmos carrying cellulose binding module (CBM) than for the lpmos without CBMs. In both cultures, the expression levels of the lpmo genes were generally higher than the levels of cellobiose dehydrogenase (CDH) encoding genes. Based on the results of this work, the oxidative cellulose cleaving enzymes of D. squalens have essential role in cellulose degrading machinery of the fungus.

Litter cellulolytic enzyme activities in alpine timberline ecotone of western Sichuan

Aims Litter constitutes the major source of organic matter entering the soil. Different litter layers reflect different phases of decomposition. The litter originated from different plant materials and decomposition phases may have a significant impact on cellulolytic enzyme activities. Our objective was to explore the effects of vegetation types and decomposition phases on cellulolytic enzyme activities during litter decomposition process in an alpine timberline ecotone at the end of snow melting. Methods The activities of three cellulolytic enzymes(β-1,4-endoglucanase, β-1,4-exoglucanase and β-1,4- glucosidase) and litter qualities(C, N, P and cellulose content) were measured in the fresh litter and fermentation layer(LF) and the humus layer(H) in alpine meadow, alpine shrub, and coniferous forest in the alpine timberline ecotone in western Sichuan. Two-way ANOVA was used for testing the main effects of vegetation types, decomposition phase and their interactions on cellulolytic enzyme activities and litter qualities. We used Spearman correlations to explore the relationships between cellulolytic enzyme activities and litter qualities of two decomposition phases. Important findings Cellulolytic enzyme activities and cellulose contents in the LF layer were significantly higher than in the H layer across all vegetation types. Two-way ANOVA results showed that decomposition phase had a more significant impact on cellulolytic enzyme activities and cellulose contents than vegetation types. Cellulolytic enzyme activities were under the control of different factors between the two decomposition stages. In the early decomposition stage, the activities of β-1,4-exoglucanase and β-1,4-glucosidase appeared to be limited by N and P contents of the substrate, while β-1,4-endoglucanase activity was mainly controlled by the cellulose content of litter. In the late decomposition stage, the activities of β-1,4-endoglucanase and β-1,4-glucosidase were mainly limited by C and N contents. According to the prediction of ecological stoichiometry theory, microbial growth is considered to be nutrient-limited on substrates with C:N 27 or C:P 186. Overall, litter C:N and C:P were greater than 27 and 186, respectively, in the study area, indicating that cellulolytic enzyme activities werelimited by litter N and P contents. In particular, the microbial biomass was limited more significantly by N and P contents in the early decomposition stage in the alpine meadow, indicating that litter quality indirectly regulates cellulolytic enzyme activities of litter decomposition process in this alpine timberline ecotone.

Exotic earthworms alter soil microbial community composition and function

Exotic earthworms can profoundly alter soil carbon (C) and nitrogen (N) dynamics in northern temperate forests, but the mechanisms explaining these responses are not well understood. We compared the soil microbial community (SMC) composition (measured as PLFAs) and enzyme activity between paired earthworm-invaded and earthworm-free plots in northern hardwood forests of New York, USA. We hypothesized that differences in SMCs and enzyme activity between plots would correspond with differences in soil C content and C:N ratios. Relative abundance of several bacterial (mostly gram-positive) PLFAs was higher and that of two fungal PLFAs was lower in earthworm compared to reference plots, largely because of earthworm incorporation of the organic horizon into mineral soil. In surface mineral soil earthworms increased arbuscular mycorrhizal fungi (AMF) and gram-positive bacterial PLFAs, and decreased fungal (mostly saprotrophic) and several bacterial (gram-negative and non-specific) PLFAs. Earthworms also increased the activities of cellulolytic relative to lignolytic enzymes in surface mineral soil, and the relationships between enzyme activities and components of the SMC suggest a substrate-mediated effect on the SMC and its metabolism of C. A highly significant relationship between components of the SMC and soil C:N also suggests that earthworms reduce soil C:N through functional and compositional shifts in the SMC. Finally, changes in AMF abundances were linked to phosphatase activity, suggesting that earthworms do not necessarily inhibit P-acquisition by AMF-associated plants in our study system. We conclude that the combined influence of earthworm-related changes in physical structure, accessibility and chemistry of organic matter, and relative abundance of certain groups of fungi and bacteria promote C metabolism, in particular by increasing the activities of cellulolytic vs. lignolytic enzymes.

Xlr1 is involved in the transcriptional control of the pentose catabolic pathway, but not hemi-cellulolytic enzymes in Magnaporthe oryzae

Magnaporthe oryzae is a fungal plant pathogen of many grasses including rice. Since arabinoxylan is one of the major components of the plant cell wall of grasses, M. oryzae is likely to degrade this polysaccharide for supporting its growth in infected leaves. D-Xylose is released from arabinoxylan by fungal depolymerising enzymes and catabolized through the pentose pathway. The expression of genes involved in these pathways is under control of the transcriptional activator XlnR/Xlr1, conserved among filamentous ascomycetes. In this study, we identified M. oryzae genes involved in the pentose catabolic pathway (PCP) and their function during infection, including the XlnR homolog, XLR1, through the phenotypic analysis of targeted null mutants. Growth of the Δxlr1 strain was reduced on D-xylose and xylan, but unaffected on L-arabinose and arabinan. A strong reduction of PCP gene expression was observed in the Δxlr1 strain on D-xylose and L-arabinose. However, there was no significant difference in xylanolytic and cellulolytic enzyme activities between the Δxlr1 mutant and the reference strain. These data demonstrate that XLR1 encodes the transcriptional activator of the PCP in M. oryzae, but does not appear to play a role in the regulation of the (hemi-) cellulolytic system in this fungus. This indicates only partial similarity in function between Xlr1 and A. niger XlnR. The deletion mutant of D-xylulose kinase encoding gene (XKI1) is clearly unable to grow on either D-xylose or L-arabinose and showed reduced growth on xylitol, L-arabitol and xylan. Δxki1 displayed an interesting molecular phenotype as it over-expressed other PCP genes as well as genes encoding (hemi-) cellulolytic enzymes. However, neither Δxlr1 nor Δxki1 showed significant differences in their pathogeny on rice and barley compared to the wild type, suggesting that D-xylose catabolism is not required for fungal growth in infected leaves.

Cellulase On-Site Production from Sugar Cane Bagasse Using Penicillium echinulatum

Penicillium echinulatum was evaluated as a cellulolytic enzyme producer in shaking flasks and bioreactor submerged culture using sugarcane bagasse as carbon source. Sodium hydroxide delignified steam-exploded pretreated bagasse (SDB) and hydrothermal pretreated bagasse had a maximum filter paper activity (FPase) of 2.4 and 2.6 FPU/mL, respectively. Delignified acid pretreated bagasse and Celufloc 200TM (CE) carbon sources displayed maximum FPase of 1.3 and 1.6 FPU/mL while in natura bagasse (INB) provided the lowest enzyme activity, ca. 0.4 FPU/mL. Measurement of surface specific area of lignocellulosic material and scanning electron microscopic images showed a possible correlation between fungal mycelia accessibility to lignocellulosic particles and obtained cellulolytic enzyme activity of fermentation broth. Fed-batch experiments performed in a controlled bioreactor attained the highest value of FPase of 3.7 FPU/mL, enzyme productivity of 25.7 FPU/L h, and enzyme yield from cellulose equal to 134 FPU/g with SDB. Enzyme hydrolysis of steam-pretreated bagasse accomplished with the obtained supernatant of fermentation broth (10 FPU/g of biomass and 5 % w/v) performed better than commercial cellulose complex. The results showed that P. echinulatum has potential to be used as an on-site enzyme platform aiming second bioethanol production from sugarcane lignocellulosic residue.