Enzyme Production Research Articles

The Utilization of Bacillus spp. as Endophytes in Enhancing Plant Resistance and Health Against Fusarium oxysporum : A Mini Review

Fusarium oxysporum is a destructive pathogen responsible for significant losses in chili (Capsicum spp.) and other crops. Traditional chemical control methods pose environmental and health risks, highlighting the need for sustainable alternatives. This review explores the role of Bacillus spp. as endophytic biocontrol agents in combating F. oxysporum. Species such as B. subtilis, B. mojavensis, and B. velezensis exhibit antifungal activity through the production of hydrolytic enzymes (e.g., glucanase, cellulase, and chitinase) and secondary metabolites like lipopeptides and antibiotics, which inhibit pathogen growth. Furthermore, these bacteria enhance plant health by inducing systemic resistance, promoting root development, and creating a rhizosphere environment unfavorable to pathogens. Their synergistic interactions with other beneficial microorganisms further bolster plant defense and productivity. Molecular and physiological analyses confirm the efficacy of Bacillus spp. in reducing disease severity and improving plant resilience. Continued research into their mechanisms and field application can support more sustainable agricultural practices.

Exploring the antivirulence potential of phenolic compounds to inhibit quorum sensing in Pseudomonas aeruginosa.

Bacteria coordinate gene expression in a cell density-dependent manner in a communication process called quorum sensing (QS). The expression of virulence factors, biofilm formation and enzyme production are QS-regulated phenotypes that can interfere in human health. Due to this importance, there is great interest in inhibiting QS, comprising an anti-virulence strategy. This work aimed to evaluate the effect of selected phenolic compounds on the inhibition of QS-regulated phenotypes in Pseudomonas aeruginosa PAO1, using concentrations that do not interfere in bacterial growth. This is one of the main premises for studying the effect of compounds on QS. Firstly, an in-silico study with the LasR and RhlR proteins of P. aeruginosa by molecular docking of 82 phenolic compounds was performed. Then, a screening with 13 selected phenolic compounds was performed, using biosensor strains P. aeruginosa lasB-gfp and P. aeruginosa rhlA-gfp, which emit fluorescence when the QS system is activated. From this assay, eight compounds were selected and evaluated for inhibition of pyocyanin, rhamnolipids, proteases, elastase, and motility. The compounds variably inhibited the evaluated virulence factors. The greatest inhibitions were observed for swarming motility, achieving inhibition rates of up to 50% for baicalein (500 µM) and curcumin (50 µM). Notably, curcumin showed satisfactory inhibition for all phenotypes even at lower concentrations (12.5 to 50 µM) compared to the other compounds (125 to 500 µM). Four compounds - rosmarinic acid, baicalein, curcumin, and resveratrol - were finally tested against biofilm formation observed by optical microscopy. This study demonstrated that phenolic compounds exhibit strong in silico binding to P. aeruginosa LasR and RhlR proteins and variably inhibit QS-regulated phenotypes in vitro. Although no biofilm inhibition was observed, future studies combining compounds and exploring molecular mechanisms are recommended. These findings highlight the biotechnological potential of phenolic compounds for future applications in the food, clinical, and pharmaceutical fields.

Optimization of Cellulases under Solid State Fermentation by Newly Isolated Fungus: An Environmentally Sustainable Approach

There exists a substantial interest in advancing the commercial production of cellulolytic enzymes. This drive is fueled by the pursuit of cost-effective substrates and energy-efficient fermentation processes, all aimed at enhancing the economic viability of enzymatic conversion of lignocellulosic biomass (LB) into bioethanol. In the present study, emphasis was given to the isolation of superior cellulase-producing fungal isolate. A comprehensive collection of 199 fungal isolates was derived from diverse soil samples, and the fungal isolate with the largest hydrolytic halos was identified as Trichoderma atroviride AD-130 through molecular analysis. The potential of cellulase production was explored and optimized during solid state fermentation (SSF) using inexpensive substrates such as Eichhornia crassipes and Municipal Solid Waste (MSW). Trichoderma atroviride AD-130 displayed the maximum cellulase production on the fifth day at pH 7.0 at a substrate-moisture ratio of 1:6 for E. crassipes and 1:2 for MSW under solid-state fermentation. The cellulases obtained were partially purified and characterized for their optimal pH (6.0, 4.5, and 5.0 for FPase, CMCase, and BGL respectively) and temperature conditions (60°C, 50°C, and 70°C for FPase, CMCase, and BGL respectively). Zymogram analysis revealed that the cellulolytic fungus T. atroviride AD-130 possessed multiple alleles for the synthesis of CMCase, which is crucial for the effective degradation of various types of lignocellulosic substrates.

Aeration promotes Proteobacteria over Firmicutes in macerated food waste resulting in superior anaerobic digestion efficiency.

Aeration is a common pretreatment method to enhance biogas production via anaerobic digestion of waste organic feedstocks such as unused food. While impacts on downstream anaerobic digestion have been intensively investigated, the consequence of aeration on the microbial community in food waste has not been characterised. Food waste has a low pH resulting from the dominance of lactic acid bacteria within the Firmicutes phylum. This excludes other phylotypes with a higher potential to hydrolyse complex biopolymers in food waste. In this study we reveal that aeration of macerated food waste results in a dramatic shift away from Firmicutes towards dominance of Proteobacteria that are better known for extracellular enzyme production. Given that hydrolysis is the rate limiting step in anaerobic digestion, this explains why aeration improves the efficiency of biogas production from food waste. The discovery that Proteobacteria dominate microbial communities in aerated food waste opens up opportunities to manipulate extracellular enzyme production through gene expression mechanisms common among Proteobacteria such as quorum sensing.

Exploring the bioeconomic potential of valorizing agroindustrial by-products in enzyme production

Exploring the bioeconomic potential of valorizing agroindustrial by-products in enzyme production

Effect of Fold-Promoting Mutation and Signal Peptide Screening on Recombinant Glucan 1,4-Alpha-maltohydrolase Secretion in Pichia pastoris.

Glucan 1,4-alpha-maltohydrolase (3.2.1.133, GMH) is an important biocatalyst in the baking industry, which could delay the retrogradation of bread and improve its cold-storage durability. In the present study, a newly cloned Thgmh was characterized and secreted by Pichia pastoris (Komagataella pastoris). After computationally assisted rational design that promotes peptide folding, the maltogenic activity in supernatant was enhanced 1.6-fold in comparison with the base strain. The signal leading sequence screening and the gene dosage increment further improved secretion by approximately 6.4-fold. The purified rationally designed ThGMHs exhibited maximal activity against soluble starch at pH 7.0 and 60℃, and maltose is the main catalytic product. In a 5-L bioreactor, conventional fed-batch fermentation resulted in 6130 U mL-1 extracellular maltogenic activity. Therefore, a promising strain for GMH production was developed, which provides a useful reference for the secretory production of other industrial enzymes.

Isolation of Actinobacteria from Date Palm Rhizosphere with Enzymatic, Antimicrobial, Antioxidant, and Protein Denaturation Inhibitory Activities

Arid ecosystems constitute a promising source of actinobacteria producing new bioactive molecules. This study aimed to explore different biological activities of actinomycetes isolated from the rhizosphere of Phoenix dactylifera L. in the Ghardaia region, Algeria. A total of 18 actinobacteria were isolated and studied for their enzymatic and antimicrobial activities. All isolates shared cellulase and catalase activity; most of them produced amylase (94%), esterase (84%), lecithinase and lipoproteins (78%), caseinase (94%), and gelatinase (72%). The isolates could coagulate (56%) or peptonize (28%) skim milk. Overall, 72% of the isolates exhibited significant antibacterial activity against at least one test bacteria, while 56% demonstrated antifungal activity against at least one test fungi. Based on enzyme production and antimicrobial activity, isolate SGI16 was selected for secondary metabolite extraction by ethyl acetate. The crude extract of SGI16 was analyzed using DPPH and BSA denaturation inhibition tests, revealing significant antioxidant power (IC50 = 7.24 ± 0.21 μg mL−1) and protein denaturation inhibitory capacity (IC50 = 492.41 ± 0.47 μg mL−1). Molecular identification based on 16S rDNA analysis showed that SGI16 belonged to the genus Streptomyces. The findings highlight that date palms’ rhizosphere actinobacteria are a valuable source of biomolecules of biotechnological interest.

Enzymatic nanomotors with chemotaxis for product-based cancer therapy

The development of an intelligent nanomotor system holds great promise for enhancing the efficiency and effectiveness of antitumor therapy. Leveraging the overexpressed substances in the tumor microenvironment as propellants and chemotactic factors for enzyme-powered nanomotors represents a versatile and compelling approach. Herein, a plasma amine oxidase (PAO)-based chemotactic nanomotor system has been successfully developed, with the ability to enzymatically produce toxic acrolein and H2O2 from the upregulated polyamines (PAs) in the tumor microenvironment for active tumor therapy. Zwitterionic polymeric nanoparticles with superior biocompatibility are synthesized, followed by PAO modification via electrostatic interactions. As expected, the resulting nanomotor system exhibits positive chemotaxis toward PAs concentration gradient. Upon reaching the tumor region, our nanomotors, actuated by the tumor microenvironmental PAs, effectively enhance diffusion and enable deep penetration into the tumor site. This leads to the induction of tumor apoptosis and simultaneous inhibition of tumor invasion and migration by decomposing PAs into toxic products. By smartly utilizing the consumption of these local chemotactic factors and their enzymatic products, our nanomotor system provides a versatile and intelligent platform for active and enhanced tumor therapy.

Isolation, characterization, and immobilization of β-galactosidase from Klebsiella michiganensis B5582Y for enhanced transgalactosylation.

β-Galactosidases are highly desirable in various biotechnological applications. However, research on those obtained from Klebsiella strains has been noticeably restricted. The present investigation centers on the isolation, purification, and characterization of a β-galactosidase enzyme derived from Klebsiella michiganensis (GALB5582Y). Additionally, the study aims to immobilize GALB5582Y onto functionalized graphene oxide (GO)-based polystyrene electrospun nanofibrous membranes (ENMs). The ultimate goal is to enhance the enzyme's transgalactosylation and catalytic efficiency, thereby expanding its range of potential applications. The GALB5582Y gene was sequenced, revealing a 3354bp sequence that encodes 1024 amino acids. This discovery provides vital information about the gene's structural arrangement. The effectiveness of functionalized graphene oxide (GO)-based engineered nanomaterials (ENMs) in immobilising GALB5582Y was confirmed using SEM, FTIR, and XRD investigations. Significant stability was reported during assessments, with the enzyme activity remaining extended. Additionally, it was shown that the enzyme was efficiently distributed across the surface of the ENM. Although there have been breakthroughs in enzyme production and immobilisation techniques, there is still room for improvement in maximizing the effectiveness of GALB5582Y immobilisation and increasing the yield of galactooligosaccharides (GOS). This calls for additional investigation and refinement.

Intestinal flow and digestive parameters of Lutzomyia longipalpis larvae.

Lutzomyia longipalpis Lutz & Neiva, 1912 (Diptera, Psychodidae), is the primary vector of Leishmania infantum Nicole, 1908, the etiological agent of American visceral leishmaniasis. During their development, sandfly larvae pass through four instars, consuming soil particles enriched with microorganisms and decomposing organic material. In numerous insect species, the intestinal epithelium not only secretes digestive enzymes and absorbs digested nutrients but also carries out additional functions, such as regulating luminal pH and facilitating the absorption or secretion of ions and water. The transport of ions and water plays a crucial role in the establishment of a countercurrent flow responsible for recycling soluble digestive enzymes. This study aimed to explore specific aspects of digestion in L. longipalpis larvae that remain poorly understood. We measured the intestinal flow within the endoperitrophic space, which varied depending on the type of diet offered to the larvae, with an average total time of 191 min. Additionally, we demonstrated the countercurrent flow in L. longipalpis larvae. Finally, we showed that the production of digestive enzymes can be modulated by nutrient availability, particularly by the amino acids in the larval diet. The higher the amino acids concentration, the higher the trypsin activity. On the other hand, the aminopeptidase activity was poorly influenced by the amino acids concentration.

AMAZON ACAI AND PINEAPPLE RESIDUES INDUCE THE ENZYME LACCASE IN Pleurotus ostreatus: APPLICATION TO BISPHENOL A BIOREMEDIATION

Bisphenol A (BPA), an endocrine disruptor used in different commercial polymers, is a persistent pollutant commonly found in effluents. Conventional wastewater treatment processes have low chemical removal efficiency and are expensive. This work evaluated BPA removal using laccase enzyme produced from white rot fungus Pleurotus ostreatus. For the enzymatic production, residues of acai berry and pineapple were used as laccase inducers. The use of natural inducers led to a high enzyme production (1139 and 1031 U mL-1 to acai and pineapple, respectively). Bisphenol A was removed to a concentration lower than LOD (limit of detection) after 4 h. The degradation mechanism of BPA occurred by oxidation of methyl to hydroxymethyl group in a propane portion, with breakdown of the aromatic ring. The developed technology brought to the scene a new green, viable methodology, using vegetable waste, adding value to these residues and bringing an alternative to the BPA treatment.

Reduction of iron porphyrin nitrate to the iron nitrosyl by H2S/thiol. studies in sublimed layers.

Hemes play key roles in enzymatic production of the mammalian gasotransmitter NO by nitric oxide synthase as well as in conversion from inorganic nitrite. In the present study, we report a hitherto unknown pathway of nitrosyl formation via thiol reduction of a iron porphyrin nitrate complex in the solid state.

Microbial adaptation and genetic modifications for enhanced remediation in low-permeability soils.

Low-permeability soils, characterized by fine texture and high clay content, pose significant challenges to traditional soil remediation techniques due to limited hydraulic conductivity, restricted nutrient flow, and reduced oxygen availability. These unique properties enable low-permeability soils to function as natural barriers in environmental protection; however, they also trap contaminants, making traditional remediation efforts challenging. This review synthesizes current knowledge on microbial adaptation and genetic engineering approaches that enhance the effectiveness of bioremediation in such environments. Key microbial adaptations, including anaerobic metabolism, extracellular enzyme production, and stress response mechanisms, allow individual microbes to adapt in low-permeability soils. Additionally, community-level strategies like microhabitat creation, biofilm formation, and functional redundancy further support microbial resilience. Advancements in genetic engineering now enable the modification of microbial traits-such as soil adhesion, nutrient utilization, and stress tolerance-to enhance bioremediation efficacy. Synthetic biology techniques further allow for the design of tailored microbial consortia that work cooperatively to degrade contaminants in complex soil matrices. This review highlights the integration of microbial and genetic engineering strategies, offering a comprehensive overview that informs current practices and guides future research in low-permeability soil remediation.

In vivo mechanism of the interaction between trimethylamine lyase expression and glycolytic pathways.

Recent studies confirmed that host-gut microbiota interactions modulate disease-linked metabolite TMA production via TMA lyase. However, microbial enzyme production mechanisms remain unclear. In the present study, we investigated the impact of dietary and intervention factors on gut microbiota, microbial gene expression, and the interplay between TMA lyase and glycolytic pathways in mice. Using 16S rRNA gene sequencing, metagenomics, and metabolomics, the gut microbiota composition and microbial functional gene expression profiles related to TMA lyase and glycolytic enzymes were determined. The results revealed that distinct diets and intervention factors altered gut microbiota, gene expression, and metabolites linked to glycine metabolism and glycolysis. Notably, an arabinoxylan-rich diet suppressed genes linked to choline, glycine, glycolysis, and TMA lyase, favoring glycine utilization via pyruvate pathways. Glycolytic inhibitors amplified these effects, mainly inhibiting pyruvate kinase. Our findings underscored the crosstalk between TMA lyase and glycolytic pathways, regulating glycine levels, and suggested avenues for targeted interventions and personalized diets to curb choline TMA lyase production.

Production of Lipase Enzyme by Marine Actinobacteria With Various pH and Temperature

Abstract: The demand for enzymes as biocatalysts in industry is very high. Research and development of different types of enzymes from different sources has started. One very important enzyme to study is the enzyme lipase. Lipase enzymes are enzymes of the hydrolase class that catalyze the hydrolysis of triglycerides to glycerol and free fatty acids. Lipases are found in a variety of sources including animals, plants, and microorganisms. Marine microorganisms, including marine actinobacteria, cannot be separated from this enzyme source's research and development process. The purpose of this study was to obtain a test protocol and optimal pH and temperature conditions for the hydrolysis reaction by lipase enzymes from marine actinobacteria. Optimal pH and temperature conditions for hydrolysis reactions by lipase enzymes from marine actinobacteria using spectrophotometry at different pH values and different temperatures of 3, 4, 5, 6, 7, 8, 9, and 10. Temperatures of 30 °C, 40 °C, 50 °C, 60 °C, 70 °C, 80 °C and 90 °C are measured at a wavelength of 405 nm. The results showed that the activity of the lipase enzyme at pH 9 with Tris-HCl buffer was the optimum pH, and the temperature of 70°C was the optimum temperature for the lipase hydrolysis reaction of marine actinobacteria.Abstrak: Kebutuhan enzim sebagai biokatalisator dalam bidang industri sangat tinggi. Berbagai macam enzim dari beragam sumber sudah mulai diteliti dan dikembangkan. Salah satu enzim yang sangat penting untuk diteliti adalah enzim lipase. Enzim lipase merupakan enzim golongan hidrolase yang mengkatalis proses hidrolisis trigliserida menjadi gliserol dan asam lemak bebas. Lipase dapat ditemukan dalam berbagai sumber seperti pada hewan, tumbuhan, dan mikroorganisme. Mikroorganisme laut tidak terlepas dari proses penelitian dan pengembangan sumber enzim ini termasuk Actinobacteria laut. Tujuan penelitian ini adalah untuk memperoleh protokol uji dan kondisi pH dan temperatur optimum reaksi hidrolisis oleh enzim lipase dari Actinobacteria laut. Kondisi pH dan temperatur optimum reaksi hidrolisis oleh enzim lipase dari Actinobacteria laut dilakukan secara kuantitatif dengan metode spektrofotometri pada variasi pH 3, 4, 5, 6, 7, 8, 9, dan 10 dan variasi suhu 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, dan 90°C diukur pada Panjang gelombang 405 nm. Hasil menunjukkan bahwa aktivitas enzim lipase pada pH 9 menggunakan buffer Tris HCl merupakan pH optimum dan temperatur 70°C merupakan temperature optimum reaksi hidrolisis enzim lipase dari Actinobacteria laut.

Cloning of glucoamylase gene from Aspergillus niger and its expression in Pichia pastoris

Glucoamylase (1,4-α-glucosidase) is a crucial commercial enzyme responsible for the conversion of starch, glycogen, and oligosaccharides into D-glucose through the hydrolysis of their non-reducing terminal glycosidic bonds. While many microorganisms, including bacteria, yeasts, and fungi, can produce glucoamylase, fungal glucoamylase is the preferred choice for industrial applications. The goal of this study was to produce the glucoamylase enzyme recombinantly in Pichia pastoris. To achieve this, the glaA gene from Aspergillus niger, responsible for encoding the glucoamylase enzyme, was cloned into a plasmid (pGAPZα-A) under the control of the GAP promoter and subsequently transferred into P. pastoris. The gene was verified through sequence analysis, while the effectiveness of transfection was validated using colony PCR and enzyme activity assays. The results demonstrated that the recombinant P. pastoris strain successfully secreted a substantial amount of glucoamylase (307.05 mg/L). The activity of the recombinant enzyme was measured at 79 U/mL.min. The enzyme exhibited robust activity over a broad range of temperatures (50-80°C) and various pH levels (pH 5-10), retaining 92-60% of its maximum activity. In conclusion, this study highlights the potential for laboratory-scale production of the glucoamylase enzyme, crucial for various industries, from a cost-effective and easily cultivable recombinant yeast strain, P. pastoris.

Optimization of Black Garlic Protein Extraction Process and Exploration of Its Properties and Functions with Enzymatic Hydrolysis Products.

This study optimized the process of extracting protein from black garlic using an alkaline dissolution and acid precipitation method through response surface methodology. The optimal extraction conditions were determined as a solid-to-liquid ratio of 1:50, an extraction time of 100 min, an extraction temperature of 30 °C, and an alkaline extraction pH of 9.0. Under these optimized conditions, the actual black garlic protein (BGP) extraction yield was 12.10% ± 0.21%, and the isoelectric point of the obtained BGP was 3.1. Subsequently, this study extracted black garlic protein under optimal conditions and subjected it to enzymatic hydrolysis using different enzymes (trypsin, pepsin, and their mixed enzymes). The functional characteristics, antioxidant activity, and hypoglycemic activity of black garlic protein before and after enzymatic hydrolysis were compared. Among the hydrolysates, the pepsin hydrolysate (BGPH-P) had the smallest particle size (188.57 ± 1.93 nm) and the highest Zeta potential (-29.93 ± 0.42 mV). Scanning electron microscopy showed that BGPH-P had the smallest and most dispersed particles. Fourier-transform infrared (FTIR) spectroscopy revealed that the dual enzymatic hydrolysis hydrolysate (BGPH-PT) exhibited the most stable structure. Compared to BGP, the hydrolysates demonstrated significantly improved solubility, water-holding capacity, and foaming ability (p < 0.05), while their emulsifying activity, emulsion stability, DPPH radical scavenging capacity, and hypoglycemic activity decreased. In summary, the BGP extracted using the optimized process demonstrated good antioxidant and hypoglycemic activities, while its enzymatic hydrolysate BGPH-P exhibited excellent solubility, water-holding capacity, and emulsifying properties, providing valuable insights for the further development of black garlic protein and its hydrolysates.

Uji Potensi Isolat Rhizobakteri Menekan Pertumbuhan Jamur Antraknosa Cabai Merah (Capsicum annum L.) secara In Vitro

The main problem in red chili production is the attack of pests and diseases, particularly anthracnose caused by the fungus Colletotrichum spp., which can reduce yields by up to 60% if not controlled. Control methods typically involve the use of fungicides; however, excessive use can lead to pathogen resistance and negative environmental impacts. Therefore, there is a need for more environmentally friendly control alternatives, such as the utilization of biocontrol agents like rhizobacteria. This study aims to evaluate the inhibitory capacity and potential of rhizobacteria in suppressing the growth of Colletotrichum sp. fungi. In the first phase, 20 rhizobacterial isolates were tested, and 5 isolates were selected for further testing in the second screening phase. The research employed a non-factorial Completely Randomized Design, consisting of 6 rhizobacterial treatments with 5 replications, resulting in a total of 30 experimental units. The best inhibitory capacity was observed in rhizobacterial isolate RB 5 (2.44%) and RB 2 (2.41%). All rhizobacterial isolates exhibited variations in colony characteristics, color, and staining results. Additionally, all tested isolates showed potential for phosphate (P) solubilization, indicated by the formation of clear zones around the rhizobacterial suspensions. The highest catalase enzyme production was observed in isolates RB 3 and RB 5. While the rhizobacterial isolate RB 3 demonstrated pathogenicity, as evidenced by the softening of potato tissue after being scratched with the isolate, it still maintained a good inhibitory effect against the growth of Colletotrichum sp.

Transcriptomic and metabolomic responses of maize under conventional and biodegradable microplastic stress

AbstractThe increasing accumulation of microplastics in agricultural soils potentially threatens crop safety and quality. However, studies regarding the molecular mechanisms underlying the effects of conventional and biodegradable microplastics on plant growth remain limited. Herein, we estimated the effects of biodegradable polybutylene adipate terephthalate, poly (butylene succinate), polylactic acid, and conventional non‐biodegradable polyethylene and polystyrene microplastics (at a concentration of 1% [w/w]) on the growth and physiological performance of maize (Zea mays L.). In addition, we studied the molecular mechanisms underlying the effects of these microplastics on maize. Exposure to microplastics induced the production of antioxidant enzymes and antioxidants at varying levels in the maize. While the maize antioxidant systems were induced against biodegradable microplastic exposure, maize photosynthesis was relatively more important for conventional microplastic treatments. Additionally, metabolomics and transcriptomic analyses revealed that the pathways of secondary metabolite biosynthesis, photosynthesis, energy metabolism, and carbohydrate metabolism were regulated by biodegradable and conventional microplastics. Specifically, microplastics induced the plant hormone signal transduction and mitogen‐activated protein kinase signaling pathways. Our results further indicated that microplastics could impact the plant through changing the soil environmental variables or altering the soil microbial communities. This study provides a molecular‐scale perspective on the responses of crops to microplastic contamination, and these findings will contribute to the ecological risk assessment of biodegradable and conventional microplastics.

Production of L-Asparaginase using Soil Isolate of Cladosporium sp. and Evaluation of its Antitumor and Antioxidant Activities

The enzyme L-asparaginase is given prime attention in healthcare settings owing to its significant therapeutic applications in oncology. The present study isolated five different species of fungi belonging to the genera Aspergillus (23.8 and 9.5%), Cladosporium (19%), Fusarium (14.3%) and Penicillium (19%) from soil samples. Screening on Czapek Dox’s medium indicated that the isolate Cladosporium sp. ASP4 exhibit higher potential of synthesizing L-asparaginase than other fungi by means of causing a halo zone measuring 22.3 mm dia. Fermentation of standard medium ingredients along with the solid substrate soya bean meal by the potential fungal isolate resulted in the production of 20.53 IU/mL of L-asparaginase. Optimization studies deciphered the carbon source sucrose, supplementary nitrogen ammonium nitrate, pH ≥8.0 and the temperature 50oC as favorable process parameters for the enzyme production. Purified L-asparaginase of the present study demonstrated moderate antineoplastic activity against the A549 lung cancer cells with the IC50 values of ≥50 IU/mL. The enzyme displayed better antioxidant property compared to those of previous studies by way of scavenging the radicals with the SC50 values of ≥250 mg/mL. These findings encourage carrying out further elaborate studies for prospective biotechnological and pharmaceutical applications of the purified enzyme.