Cellulase Enzyme Research Articles

OPTIMIZATION OF CARBOXY METHYL CELLULASE PRODUCTION BY CLADOSPORIUM SP. WITH FOOD WASTE IN BAGASSE SOLID STATE FERMENTATION USING RESPONSE SURFACE METHODOLOGY

Enzyme production costs should be reduced by using inexpensively available substrates. In this, we determine the effects of various medium components and food processing waste on Cladosporium sp. production of CMCase in SSF. Five factors were tested with a three-level Box-Wilson factorial, central composite design (CCD) to determine how they impacted Cladosporium sp. NCIM 901's ability to produce CMCase. For the model, the R2 (coefficient of determination) value was 0.9349. Based on the process findings, the optimal medium for higher enzyme synthesis had wheat bran (4 g/L), cacl2 (2 g/L), urea (7.5 g/L) and rice bran (8.76 g/L), with the medium's original pH at 5.5. The model's accuracy is confirmed by the experimental result of CMCase, which is 23.42 U/gds and equivalent to the expected yield of 22.31 U/gds. The accuracy of the current experimental design has been verified by experimental validation. The enzyme's properties indicated that its greatest activity occurred at pH 5.5 and that its ideal temperature was about 50 °C. The enzyme is more active toward CMC and less active toward filter paper. CaCl2 and MgCl2 increased the enzyme's activity. Enzymatic hydrolysis of waste-derived polysaccharides into simple sugars is necessary for the industrial manufacture of bioethanol. The efficacy of the produced Cladosporium sp. cellulase (CMCase) enzyme to saccharify pearl millet grass results shows that H2SO4 treated substrate gave more fermentable sugars compared to the untreated substrate. Results suggest that the indigenous produced-CMCase can be efficiently used for lignocellulosic biomass saccharification (55.40%) compared to the untreated sample (37.10%) in 48 h, which will eventually reduce the cost utilized for purchasing commercial enzymes.

Iskanje in določanje bakterij, ki so sposobne razgraditi celulozo s pomočjo IAA kot potencialnih pospeševalcev rasti rastlin

Strains with both straw degradation and plant growth promotion ability were selected from the cultivated soil in Bac Kan, Vietnam to solve the problems of poor soil microbial diversity status, weak corrosion promotion effect, and poor crop growth caused by fungal rot diseases. Among seventeen bacteria isolated, strain NR1 presented the highest value for cellulase enzyme activity (Hydrolysis index = 24.8 mm), and IAA production (20.15 mg l-1), and was identified as Bacillus amyloliquefaciens Priest et al., 1987. Inoculation with NR1 significantly increased the rot promotion rate of straw under liquid fermentation by 54.71 % compared with the control and increased the root length and average diameter, and SPAD value of maize under soil culture by 18.3 %, 22.0 %, and 5.24 % respectively (p < 0.05). In addition, fertilizing 8 or 9 tons of NR1-degraded compost fertilizer per hectare had the best effect on the growth, development, and productivity of the L14 peanut variety. These results suggest strain NR1 could be used to produce multi-functional humus, accelerate the decomposition of straw in the cultivated soil, and promote crop growth.

Cellobionate production from sodium hydroxide pretreated wheat straw by engineered Neurospora crassa HL10

This study investigated cellobionate production from a lignocellulosic substrate using Neurospora crassa HL10. Utilizing NaOH-pretreated wheat straw as the substrate obviated the need for an exogenous redox mediator addition, as lignin contained in the pretreated wheat served as a natural mediator. The low laccase production by N. crassa HL10 on pretreated wheat straw caused slow cellobionate production, and exogenous laccase addition accelerated the process. Cycloheximide induced substantial laccase production in N. crassa HL10, enabling the strain to yield approximately 57 mM cellobionate from pretreated wheat straw (equivalent to 20 g/L cellulose), shortening the conversion time from 8 to 6 days. About 92% of the cellulose contained in the pretreated wheat straw is converted to cellobionate. In contrast to existing methods requiring pure cellobiose or cellulase enzymes, this process efficiently converts a low-cost feedstock into cellobionate at a high yield without enzyme or redox mediator supplementation.

Bioethanol production from coconut husk using DES-NADES pretreatment and enzymatic hydrolysis method

Bioethanol is an alternative fuel produced during biomass fermentation. Among the available biomass resources for bioethanol production, coconut husk is an interesting raw material due to its high cellulose content. This study aims to evaluate the coconut husk conversion into bioethanol using DES (ChCl:MEA) and NADES (Be:La) solvents for the delignification process. The molar ratios of HBA and HBD of the two solvents were varied (1:4, 1:6, 1:8) at 2, 4, 6, and 8 h for each ratio. The delignified samples were hydrolyzed using the 5% (w/w) of cellulase enzymes and fermented for seven days using Saccharomyces cerevisiae. The results showed that DES (ChCl:MEA) and NADES (Be:La) could remove lignin at about 60.53% and 65.81%. The glucose content was obtained at 0.5% (brix) by both solvents and bioethanol content was obtained at 13.9% and 14% (v/v) for DES and NADES, respectively.

Enhancement in sugar extraction from Pistia stratiotes through statistical optimization of alkaline pre-treatment and enzymatic hydrolysis

In lignocellulosic biomass, reducing structural recalcitrance and enhancing hydrolysis efficiency are crucial factors for increasing fermentable sugars and the production of valuable products. This biomass substrate comprises lignin, hemicellulose, and cellulose. In this study, response surface methodology was employed to optimise alkaline pre-treatment followed by enzymatic hydrolysis, aiming to enhance the saccharification of Pistia stratiotes. The NaOH concentration during pre-treatment significantly influenced the delignification process, resulting in increased cellulose content. The highest cellulose content was achieved with 2.47% NaOH at 120 °C for 60 min, leading to enhanced cell porosity and facilitating greater enzyme saccharification accessibility. Under these optimized conditions, the sample exhibited a 51.66% cellulose content. The physicochemical characteristics of the cellulose obtained after pre-treatment were analysed using SEM, FTIR, and TGA. After enzymatic hydrolysis of the cellulose with a commercially available cellulase enzyme, 31.06 g/L of reduced sugar was produced after 72 h. This study demonstrates that alkaline pre-treatment of Pistia stratiotes significantly increased its cellulose content, leading to a higher sugar yield during enzymatic hydrolysis.

Extremozymes and compatible solute production potential of halophilic and halotolerant bacteria isolated from crop rhizospheric soils of Southwest Saurashtra Gujarat

Halophiles are one of the classes of extremophilic microorganisms that can flourish in environments with very high salt concentrations. In this study, fifteen bacterial strains isolated from various crop rhizospheric soils of agricultural fields along the Southwest coastline of Saurashtra, Gujarat, and identified by 16S rRNA gene sequencing as Halomonas pacifica, H. stenophila, H. salifodinae, H. binhaiensis, Oceanobacillus oncorhynchi, and Bacillus paralicheniformis were investigated for their potentiality to produce extremozymes and compatible solute. The isolates showed the production of halophilic protease, cellulase, and chitinase enzymes ranging from 6.90 to 35.38, 0.004–0.042, and 0.097–0.550 U ml−1, respectively. The production of ectoine-compatible solute ranged from 0.01 to 3.17 mg l−1. Furthermore, the investigation of the ectoine-compatible solute production at the molecular level by PCR showed the presence of the ectoine synthase gene responsible for its biosynthesis in the isolates. Besides, it also showed the presence of glycine betaine biosynthetic gene betaine aldehyde dehydrogenase in the isolates. The compatible solute production by these isolates may be linked to their ability to produce extremozymes under saline conditions, which could protect them from salt-induced denaturation, potentially enhancing their stability and activity. This correlation warrants further investigation.

Optimization, gene cloning, expression, and molecular docking insights for enhanced cellulase enzyme production by Bacillus amyloliquefaciens strain elh1

BackgroundIn this study, we isolated a cellulase-producing bacterium, Bacillus amyloliquefaciens strain elh, from rice peel. We employed two optimization methods to enhance the yield of cellulase. Firstly, we utilized a one-variable-at-a-time (OVAT) approach to evaluate the impact of individual physical and chemical parameters. Subsequently, we employed response surface methodology (RSM) to investigate the interactions among these factors. We heterologously expressed the cellulase encoding gene using a cloning vectorin E. coli DH5α. Moreover, we conducted in silico molecular docking analysis to analyze the interaction between cellulase and carboxymethyl cellulose as a substrate.ResultsThe bacterial isolate eh1 exhibited an initial cellulase activity of 0.141 ± 0.077 U/ml when cultured in a specific medium, namely Basic Liquid Media (BLM), with rice peel as a substrate. This strain was identified as Bacillus amyloliquefaciens strain elh1 through 16S rRNA sequencing, assigned the accession number OR920278 in GenBank. The optimal incubation time was found to be 72 h of fermentation. Urea was identified as the most suitable nitrogen source, and dextrose as the optimal sugar, resulting in a production increase to 5.04 ± 0.120 U/ml. The peak activity of cellulase reached 14.04 ± 0.42 U/ml utilizing statistical optimization using Response Surface Methodology (RSM). This process comprised an initial screening utilizing the Plackett–Burman design and further refinement employing the BOX -Behnken Design. The gene responsible for cellulase production, egl, was effectively cloned and expressed in E. coli DH5α. The transformed cells exhibited a cellulase activity of 22.3 ± 0.24 U/ml. The egl gene sequence was deposited in GenBank with the accession number PP194445. In silico molecular docking revealed that the two hydroxyl groups of carboxymethyl cellulose bind to the residues of Glu169 inside the binding pocket of the CMCase. This interaction forms two hydrogen bonds, with an affinity score of −5.71.ConclusionsOptimization of cultural conditions significantly enhances the yield of cellulase enzyme when compared to unoptimized culturing conditions. Additionally, heterologous expression of egl gene showed that the recombinant form of the cellulase is active and that a valid expression system can contribute to a better yield of the enzyme.

Isolation and Characterization of Aeromonas taiwanensis Strain for Simultaneous Production of Cellulase, Amylase, Pectinase, and Protease Enzymes

Isolation and Characterization of Aeromonas taiwanensis Strain for Simultaneous Production of Cellulase, Amylase, Pectinase, and Protease Enzymes

Microbial Biodiversity of Cryoconites in Palandöken Mountain and Culturing Cold-Adapted Enzyme Producers

Palandöken Mountain is among the highest and coldest ecosystems of Türkiye, and knowledge on its microbial diversity has not been adequately characterized. The aim of the present study is to expand the current understanding of microbial diversity in Palandöken Mountain. Sampling studies were carried out at in winter an altitude of 2400–2900 m. A next-generation sequencing (NGS) based culture-independent approach was employed for elucidating the community structure in the 8 cryoconite samples using 16S and 23S rRNA gene sequencing. In addition, biotechnologically important cold-adapted enzyme producers were isolated and identified by employing conventional culture-dependent studies. High-throughput sequencing analysis revealed the presence of 36 bacterial and 2 fungal phyla. The bacterial communities of 8 samples were dominated by Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes and Acidobacteria. The fungal communities were dominated by Ascomycota and Basidiomycota. 2900 different taxonomic group for bacterial diversity and 51 for fungal diversity were reported in the present study. Furthermore, 25 cold-adapted enzyme-producers were isolated and identified in the culture-dependent studies. Each selected isolate was capable of producing at least one of the enzymes amylase, protease, lipase, cellulase and xylanase isolates were assigned to Aspergillus, Bacillus, Cladosporium, Dioszegia, Flavobacterium, Fusarium, Holtermanniella, Naganishia, Polaromonas, Protomyces, Pseudomonas Rhodosporidiobolus, Vishniacozyma and Yarrowia genera. Consequently, this is the first geomicrobiological investigation that describes bacterial and fungal diversity of cryoconites from Palandöken Mountain using an NGS-based culture-independent approach. Additionally, cultured cold-adapted enzyme producer isolates of the present study have a valuable potential for the development of new biotechnological tools for various industrial applications.

Phyco-biosynthesis of Chlorella-CuO-NPs and its Immobilization on Polyester/Cotton Blended Textile Waste Activated by Cellulase Enzymes for Application as Wastewater Disinfection Filter

Phyco-biosynthesis of Chlorella-CuO-NPs and its Immobilization on Polyester/Cotton Blended Textile Waste Activated by Cellulase Enzymes for Application as Wastewater Disinfection Filter

The Effect of Biofertilizers and Seaweed Extract on the Activity of Cellulase Enzyme and β-Glucosidase in Soil Planted with Broccoli Plants

The Effect of Biofertilizers and Seaweed Extract on the Activity of Cellulase Enzyme and β-Glucosidase in Soil Planted with Broccoli Plants

Effects of Fungal Solid-State Fermentation on the Profile of Phenolic Compounds and on the Nutritional Properties of Grape Pomace.

Grape pomace (GP) is considered a natural source of bioactive compounds. To improve the extractability of bioactive compounds, in this work, GP was biologically treated for 15 days with the white-rot fungus Trametes versicolor in laboratory jars and a tray bioreactor under solid-state fermentation (SSF) conditions. During SSF, the activity of lignolytic (laccase and manganese peroxidase) and hydrolytic (xylanase, cellulase, β-glucosidase, and invertase) enzymes was measured, with the activities of laccase (2.66 U/gdb in jars and 0.96 U/gdb in the bioreactor) and xylanase (346.04 U/gdb in jars and 200.65 U/gdb in the bioreactor) being the highest. The effect of the complex enzyme system was reflected in the changes in the chemical composition of GP with increasing ash, crude protein, and free fat content: 28%, 10%, and 17% in the laboratory jars, and 29%, 11%, and 7% in the bioreactor, respectively. In addition, the biological treatment improved the extractability of 13 individual phenolic compounds. Therefore, the applied SSF technique represents an effective strategy to improve the profile of phenolic compounds and the nutritional composition of GP, promoting their valorization and opening the door for potential applications in the food industry and other sectors.

Mechanism of Brevibacillus brevis strain TR-4 against leaf disease of Photinia×fraseri Dress.

Colletotrichum species are among the most common pathogens in agriculture and forestry, and their control is urgently needed. In this study, a total of 68 strains of biocontrol bacteria were isolated and identified from Photinia × fraseri rhizosphere soil. The isolates were identified as Brevibacillus brevis by 16S rRNA. The inhibitory effect of TR-4 on Colletotrichum was confirmed by an in vitro antagonistic experiment. The inhibitory effect of TR-4 was 98% at a concentration of 10 µl/ml bacterial solution, protection of the plant and inhibition of C. siamense was evident. Moreover, the secretion of cellulase and chitosan enzymes in the TR-4 fermentation liquid cultured for three days was 9.07 mol/L and 2.15 µl/mol, respectively. Scanning electron microscopy and transmission electron microscopy confirmed that TR-4 destroyed the cell wall of C. siamense, resulting in leakage of the cell contents, thus weakening the pathogenicity of the bacteria.

Enzymes and biosurfactants of industrial interest produced by culturable fungi present in sediments of BoeckellaLake, Hope Bay, north-east Antarctic Peninsula.

This study characterized cultivable fungi present in sediments obtained from BoeckellaLake, Hope Bay, in the north-east of the Antarctic Peninsula, and evaluated their production of enzymes and biosurfactants of potential industrial interest. A total of 116 fungal isolates were obtained, which were classified into 16 genera within the phyla Ascomycota, Basidiomycota and Mortierellomycota, in rank. The most abundant genera of filamentous fungi included Pseudogymnoascus, Pseudeurotium and Antarctomyces; for yeasts, Thelebolales and Naganishia taxa were dominant. Overall, the lake sediments exhibited high fungal diversity and moderate richness and dominance. The enzymes esterase, cellulase and protease were the most abundantly produced by these fungi. Ramgea cf. ozimecii, Holtermanniella wattica, Leucosporidium creatinivorum, Leucosporidium sp., Mrakia blollopis, Naganishia sp. and Phenoliferia sp. displayed enzymatic index > 2. Fourteen isolates of filamentous fungi demonstrated an Emulsification Index 24% (EI24%) ≥ 50%; among them, three isolates of A. psychrotrophicus showed an EI24% > 80%. BoeckellaLake itself is in the process of drying out due to the impact of regional climate change, and may be lost completely in approaching decades, therefore hosts a threatened community of cultivable fungi that produce important biomolecules with potential application in biotechnological processes.

Interaction between Enzymatic Detergent and Textile Metals/Metal Oxide Nanoparticles

aims: This research examines the interactions between metal (Ag) and metal oxide (TiO2 and ZnO) nanoparticles and amylase, cellulase, protease, and lipase as detergent enzymes and their impacts on enzyme activity. background: Nanoparticles are used in industrial products, such as textiles, to induce novel properties, such as antibacterial, antistatic, UV blocking, self-cleaning properties, wrinkle resistance, and water and oil repellent. On the other hand, using enzymes (protease, lipase, amylase, and cellulase) is widespread in detergent industries for washing conditions. objective: The interactions between metal (Ag) and metal oxide (TiO2 and ZnO) nanoparticles and detergent enzymes method: Using a central composite design, a total of 320 experiments under different conditions were conducted to determine the extent of change in enzyme activity. result: Results indicate that lipase had the lowest activity under interaction with silver nanoparticles, while cellulase and protease were most affected by interactions with AgNPs and a-TiO2. The surface response of the examined parameters showed the most effect from the interaction time and temperature and the enzyme/nanoparticle ratio and temperature parameters. conclusion: Further investigations into the effects of nanomaterials and enzymes on the physiology and chemistry of surfaces are suggested. This research results demonstrate that in nanoparticle and enzyme interface, physical, chemical, and biological difference is made in both. The outcome can be used to improve nanoparticle and detergent enzyme interactions in washing surroundings, aiming to retain nanoparticles and enzyme traits. other: -

Comparing Sensorial Comfort Properties of Cotton, Cotton/Linen and Linen Knitted Fabrics

ABSTRACT Linen fibers exhibit good moisture management properties and can effectively conduct heat. However, there are limited studies on the sensorial comfort properties of linen knitted fabrics. In this study, the sensorial comfort properties of optical bleached, different enzyme washed and softened 100% cotton, 50% cotton/50% linen, and 100% linen fabrics were investigated using the Kawabata KES-FB evaluation system. The results showed that the tensile resilience and compression linearity values of the 100% linen fabrics were higher than those of other fabrics. The compression energy and compressional resilience of the 50% cotton- 50% linen fabrics were higher than those of the other fabrics. The surface friction value of 100% linen fabrics can be improved by cellulase enzyme treatment. Pectinase enzyme treated fabrics showed higher smoothness values than cellulase enzyme treated 100% linen fabrics.

Effect of Enzyme Supplementation on Nutrient Utilization and Production Performance in Lactating Murrah Buffaloes

Background: Enzyme stimulates metabolic and digestive processes leading to increased feed intake and improved nutrient availability in animals. This study was conducted to investigate the effect of enzyme supplementation on nutrient utilization and production performance in lactating Murrah Buffaloes during winter season. Methods: Twenty four lactating Murrah buffaloes were selected for the study from Livestock Research Centre, Sardar Vallabhbhai Patel University of Agriculture and Technology, Modipuram, Meerut (UP). All the selected animals were randomly allocated into four treatment groups having six animals in each group on the basis of body weight, lactation length and milk yield. The daily nutrient requirements of lactating Murrah buffaloes in term of DM, DCP and TDN were provided as per the feeding standards of Indian Council of Agricultural Research, New Delhi (2013). Total Mixed Ration for buffaloes was prepared by mixing of roughage and concentrates in the ratio of 60:40 after grinding/chaffing. Roughage part was consisting of available winter season green fodder e.g. oat and wheat straw available at research centre. Feeding trial was planned into four treatment groups viz. T1 (Basal diet without enzyme supplementation), T2 (Basal diet with supplementation of Xylanase @ 800000 IU), T3 (Basal diet with supplementation of Cellulase @ 240000 IU) and T4 (Basal diet with supplementation of Xylanase @ 800000 IU and Cellulase @ 240000 IU). Result: From the present study, it was observed that the average milk yield (5.34 litre/day) was higher in T4 as compared to other treatment groups, but the difference was non-significant. The average milk fat percent was recorded as 7.52, 7.58. 7.60 and 7.65 in T1, T2, T3 and T4 treatment groups, respectively. Although the results of T4 group animals were comparatively higher in all major milk constituents the differences were found to be statistically non-significant. The average DM intake (kg/d, kg/100 kg BW and g/kg W0.75) and CP intake (g/d, g/100 kg BW) were almost similar in all treatment groups. The TDN intake (kg/d, kg/100 kg BW and g/kg W0.75) was significantly (P less than 0.05) higher in enzyme supplemented groups. The digestibility coefficient of CP, CF, NDF and ADF were higher (P less than 0.05) in buffaloes of T4 over control (T1) group. So, it was concluded from the above study that the milk production and milk composition could be improved with the combine use of Xylanase and Cellulase enzyme in murrah buffaloes due to improved CP and fibre digestibility.

Biocontrol of Fusarium head blight in rice using Bacillus velezensis JCK-7158.

Fusarium head blight (FHB) is a destructive disease caused by several species of Fusarium, such as Fusarium graminearum and F. asiaticum. FHB affects cereal crops, including wheat, barley, and rice, worldwide. Fusarium-infected kernels not only cause reduced yields but also cause quality loss by producing mycotoxins, such as trichothecenes and zearalenone, which are toxic to animals and humans. For decades, chemical fungicides have been used to control FHB because of their convenience and high control efficacy. However, the prolonged use of chemical fungicides has caused adverse effects, including the emergence of drug resistance to pathogens and environmental pollution. Biological control is considered one of the most promising alternatives to chemicals and can be used for integrated management of FHB due to the rare possibility of environment pollution and reduced health risks. In this study, Bacillus velezensis JCK-7158 isolated from rice was selected as an ecofriendly alternative to chemical fungicides for the management of FHB. JCK-7158 produced the extracellular enzymes protease, chitinase, gelatinase, and cellulase; the plant growth hormone indole-3-acetic acid; and the 2,3-butanediol precursor acetoin. Moreover, JCK-7158 exhibited broad antagonistic activity against various phytopathogenic fungi and produced iturin A, surfactin, and volatile substances as active antifungal compounds. It also enhanced the expression of PR1, a known induced resistance marker gene, in transgenic Arabidopsis plants expressing β-glucuronidase (GUS) fused with the PR1 promoter. Under greenhouse conditions, treatments with the culture broth and suspension concentrate formulation of JCK-7158 at a 1,000-fold dilution inhibited the development of FHB by 50 and 66%, respectively. In a field experiment, treatment with the suspension concentrate formulation of JCK-7158 at a 1,000-fold dilution effectively controlled the development of FHB with a control value of 55% and reduced the production of the mycotoxin nivalenol by 40%. Interestingly, treatment with JCK-7158 enhanced the expression of plant defense-related genes in salicylic acid, jasmonic acid, ethylene, and reactive oxygen species (ROS) signaling pathways before and after FHB pathogen inoculation. Taken together, our findings support that JCK-7158 has the potential to serve as a new biocontrol agent for the management of FHB.

Digestive Enzyme Activities in Mussel Mytilus californianus Endure Acute Heat Exposure in Air.

The mussel Mytilus californianus is an ecosystem engineer forming beds along the coastlines of Northeastern Pacific shores. As sessile organisms, they modulate their energy balance through valve movements, feeding, and digestive functionality. A recent study observed that activity of the digestive enzyme cellulase was higher than predicted in mussels high on the shore, where temperatures are characteristically high and food availability is limited compared to low-shore habitats. In the current study, we predicted that this scavenging behavior is induced to mitigate energy losses related to heat-shock responses-that cellulase and amylase will display hyperactivity for limited recourses in the face of aerial heating. In the laboratory, we acclimated mussels to three complex diets that differed in starch and cellulose composition, followed by two acute heat shocks (+8°C) in the laboratory. Results showed no hyperactivity of amylase and cellulase in heated mussels. These results differ from previous studies that showed lowered amylase activity following heat acclimation. This difference in amylase activity across heat-stress exposure time is important when analyzing mussel bed disturbances following heat waves that compromise energy balance or cause death within adult populations.

Optimization of Fermentation Conditions for Cellulase Production by Trichoderma harzianum PK5 Obtained from Decaying Palm Kernel Cake

Study’s Novelty/ Excerpt This study successfully optimized cellulase production by Trichoderma harzianum PK5 using the One Factor at a Time (OFAT) approach, identifying copra meal and KNO3 as the best carbon and nitrogen sources. Optimal conditions included a pH of 4.0, 125% moisture concentration, 8% inoculum size, 30°C temperature, and a 7-day incubation period, yielding a high enzyme titre of 252.54±7.73 U/gds in solid-state fermentation. These findings highlight the potential of T. harzianum PK5 as a cost-effective source of cellulases for industrial applications. Full Abstract Cellulases are considered to be among the most important enzymes in the commercial market and in various industries. Their applications are widespread, leading to increased demand and high associated costs. This necessitates the search for more cost-effective cellulases from microorganisms. Therefore, the aim of this study was to optimize cellulase production by Trichoderma harzianum PK5 using the One Factor at a Time (OFAT) approach. The effects of carbon, nitrogen, and various environmental factors were studied in both submerged and solid-state fermentation setups by adjusting one factor at a time based on the optimal conditions established from the previous condition. Copra meal and KNO3 were identified as the best complex carbon and nitrogen sources, respectively, for cellulase production by Trichoderma harzianum PK5. The optimal pH of 4.0, moisture concentration of 125% (v/w), inoculum size of 8%, temperature of 30°C, and an incubation time of 7 days were determined as the optimal conditions for cellulase production by this isolate, resulting in an enzyme titre of 252.54±7.73 U/gds in solid-state fermentation. It was found that cellulase enzyme production by the isolate was constitutive. In conclusion, cellulase production by T. harzianum PK5 was significantly optimized using the OFAT approach