Chitinase Production Research Articles

Insights on fungal solid-state fermentation for waste valorization: Conidia and chitinase production in different reactor configurations

Different reactor configurations are paired with a wide variety of agro-industrial wastes of different biodegradability to produce fungal conidia by solid-state fermentation.This work presents a preliminary comparative study between packed-bed and tray reactor configurations to produce Beauveria bassiana and Trichoderma harzianum conidia using two different substrates in terms of biodegradability: rice husk or beer draff complemented with wood chips. Conidia production, mean temperature and respiration indexes have been analysed in most of the presented reactor configurations. Both strains showed higher conidia production when using beer draff complemented with wood chips as substrate due to the use of a mixture as substrate. When working with beer draff, chitinase analyses obtained similar profiles in both strains but higher overall values using TH. Conidia and chitinase production maximums were not achieved at the same time, having 2–3 days of difference depending on the strain. No significant differences in mean temperature were shown between most of the performed fermentations. As a result of the present work, further scaling of both packed bed and tray configurations using beer draff and wood chips to produce BB or TH conidia would be advisable. More experiments should be performed to optimize both conidia and chitinase productions to enhance the quality of the final product.

Purification and characterization of chitinase produced by thermophilic fungi Thermomyces lanuginosus

Background In the past few years, the production of shrimp shell waste from the seafood processing industries has confronted a significant surge. Furthermore, insignificant dumping of waste has dangerous effects on both nature and human well-being. This marine waste contains a huge quantity of chitin which has several applications in different fields. The chitinase enzyme can achieve degradation of chitin, and the chitin itself can be used as the substrate as well for production of chitinase. In the current study, the chitinase enzyme was produced by Thermomyces lanuginosus. The extracellular chitinase was purified from crude extract using ammonium sulfate precipitation followed by DEAE-cellulose ion-exchange chromatography and Sephadex G-100 gel filtration chromatography. The stability and activity of chitinase with different pH, temperature, different times for a reaction, in the presence of different metal ions, and different concentration of enzyme and substrate were analyzed. Result The chitinase activity was found to be highest at pH 6.5, 50 °C, and 60 min after the reaction began. and the chitinase showed the highest activity and stability in the presence of β-mercaptoethanol (ME). The SDS-PAGE of denatured purified chitinase showed a protein band of 18 kDa. Conclusion The characterization study concludes that Cu2+, Hg2+, and EDTA have an inhibitory effect on chitinase activity, whereas β-ME acts as an activator for chitinase activity. The utilization of chitin to produce chitinase and the degradation of chitin using that chitinase enzyme would be an opportunity for bioremediation of shrimp shell waste.

Lactic acid bacteria: economic propagation, chitinases activity, and enhancing viability by gel encapsulation

Background In a previous study, a lactic acid bacterium, Enterococcus faecium, was locally isolated from Egyptian soil and its ability to inhibit the growth of a test phytopathogen was proven.Objective The study was performed to assess the ability of the tested strain to grow on different media. The produced antifungal agent was investigated. Finally, the strain was encapsulated within different biopolymers to increase its viability.Materials and methods Several byproducts were tested and compared with the standard De Man-Rogosa-Sharpe medium. The antifungal activity was tested using the poisoned food technique. Chromatographic analysis of the fermentation medium was performed using high-performance liquid chromatography. Production of chitinase was confirmed by cultivating the test strain on chitin and estimating the amount of reducing sugars using the Somogyi method. The E. faecium cells were also encapsulated within soy protein isolate-alginate beads, gellan gum discs, and carboxymethyl cellulose beads.Results and conclusion The strain was able to grow on all of the tested byproducts and exerted a potent antifungal activity against Fusarium solani, especially when a very economic medium, mainly composed of whey, was used. High-performance liquid chromatography results confirmed the production of a number of organic acids that contributed in the inhibition of the fungal growth. The study also proved the production of chitinase enzymes, which apparently altered the chitinous layer present in the cell wall of F. solani, causing disintegration of the fungal cells. It was also shown that encapsulation of E. faecium increased its viability in soil as compared with the free uncapsulated strain.

Chitinase Activity by Chitin Degrading Strain (Bacillus Salmalaya) in Shrimp Waste

Introduction: Chitin degradation by chitinase enzyme can be used on a large scale for bioremediation of seafood waste and is environmentally friendly. Objective: The main aim of this study was to screen the potential of the strain as a chitin degrading agent. Methods: In this present study, the cell-free supernatant of Bacillus salmalaya was determined for its protein concentration. Bacillus salmalaya139SI was isolated from agricultural soil and it was identified by staining technique and colony morphology. The production of chitinase by Bacillus salmalaya139SI was optimized under different concentrations of substrate, pH and temperature. Results: Strain 139SI exhibited strong hemolytic activity and the crude protein concentration of Bacillus salmalaya was 84.09mg/ mL with OD value 0.462. Strain 139SI were also screened on colloidal chitin agar medium supplemented with mineral salt. Chikinase production was determined by clear zones of hydrolysis produced after 7 days of incubation at 37°C. The maximum chitinase production was observed in Brain Heart Infusion broth supplemented with 1.0% colloidal chitin at pH 7 and temperature 35°C after four days of incubation. Chitinase activity was observed when high concentrations of crude extract of 139SI able to degrade shrimp shell by showing the degradation zone at day 4. Conclusion: From the results, we concluded that the Bacillus salmalayahas potential to be a biofunctional chitinase that could degrade complex polysaccharides present in the organic wastes and applicable in cleaning the environment.

Plant-Growth-Promoting Traits of Bacillus Species Associated with Quinoa (Chenopodium quinoa) and Lambsquarters (Chenopodium album)

As quinoa (Chenopodium quinoa Willd.) production expands globally, methods to mitigate abiotic and biotic production constraints are needed. One potential method to improve quinoa productivity is through the use of plant-growth-promoting microbes. In this study, we examined Chenopodium-associated Bacillus populations from Bolivia, Ecuador, and the United States for their ability to solubilize phosphate, antagonize Fusarium oxysporum, and produce phytase, indole acetic acid (IAA), and chitinase. The majority (99%) of isolates screened (n = 496) exhibited one or more traits related to plant growth promotion. Phosphate solubilization was observed in 75%, phytase production in 89%, IAA production in 44%, chitinase production in 13%, and antagonism to F. oxysporum in 8% of screened isolates. Some species or species groups screened were more likely than others to exhibit certain traits, such as B. megaterium, B. pumilus group, and B. subtilis group for phosphate solubilization and B. cereus group for chitinase production. This study indicates that many Chenopodium-associated Bacillus isolates exhibit plant-growth-promoting traits and could be studied further to promote quinoa production.

Production and Partial Purification of Thermostable Chitinase from Bacillus cereus BSH-4 and its Antifungal Activity

Production and Partial Purification of Thermostable Chitinase from Bacillus cereus BSH-4 and its Antifungal Activity

Utilization of Agro-Industrial Waste for Chitinase Production by Locally Isolated Bacillus subtilis using Response Surface Methodology

Utilization of Agro-Industrial Waste for Chitinase Production by Locally Isolated Bacillus subtilis using Response Surface Methodology

The Interactions among Isolates of Lactiplantibacillus plantarum and Dairy Yeast Contaminants: Towards Biocontrol Applications

Yeast diversity in the cheese manufacturing process and in the cheeses themselves includes indispensable species for the production of specific cheeses and undesired species that cause cheese defects and spoilage. The control of yeast contaminants is problematic due to limitations in sanitation methods and chemicals used in the food industry. The utilisation of lactic acid bacteria and their antifungal products is intensively studied. Lactiplantibacillus plantarum is one of the most frequently studied species producing a wide spectrum of bioactive by-products. In the present study, twenty strains of L. plantarum from four sources were tested against 25 species of yeast isolated from cheeses, brines, and dairy environments. The functional traits of L. plantarum strains, such as the presence of class 2a bacteriocin and chitinase genes and in vitro production of organic acids, were evaluated. The extracellular production of bioactive peptides and proteins was tested using proteomic methods. Antifungal activity against yeast was screened using in vitro tests. Testing of antifungal activity on artificial media and reconstituted milk showed significant variability within the strains of L. plantarum and its group of origin. Strains from sourdoughs (CCDM 3018, K19-3) and raw cheese (L12, L24, L32) strongly inhibited the highest number of yeast strains on medium with reconstituted milk. These strains showed a consistent spectrum of genes belonging to class 2a bacteriocins, the gene of chitinase and its extracellular product 9 LACO Chitin-binding protein. Strain CCDM 3018 with the spectrum of class 2a bacteriocin gene, chitinase and significant production of lactic acid in all media performed significant antifungal effects in artificial and reconstituted milk-based media.

Exploring new marine bacterial species, Alcaligenes faecalis Alca F2018 valued for bioconversion of shrimp chitin to chitosan for concomitant biotechnological applications

The exploitation of chitinous materials seems to be an infinite treasure. To this end, using shellfish waste as the sole carbon/nitrogen source solves environmental challenges while lowering microbial chitinase production costs. Bioconversion of shellfish chitin wastes such as shrimp shells has recently been investigated for the production of enzymes and bioactive materials in order to maximize the utilization of chitin-containing seafood processing wastes. In this study, the bioconversion of chitin to chitosan by Alcaligenes faecalis Alca F2018 revealed the highest chitin deacetylase (CDA) activity of 40.6 U/μg. The resulted low Km and high Vmax values explain the high affinity of the purified CDA to the p-nitroacetanilide substrate. CDA with a molecular weight of 66 KDa was purified from F2018 strain, with a 14.5% yield. FT-IR revealed distinct chitosan peaks and XRD revealed that chitosan samples had lower crystallinity than chitin. TGA analysis revealed that the recovered chitosan samples were more thermally stable. The deacetylation degree percentages of the produced chitosan are in the same range as that of the commercial chitosan, suggesting the promising potential of A. faecalis Alca F2018 to utilize shrimp shells in their raw form in the fermentation media based on its CDA enzyme activity.

Impact of Pseudomonas spp. on Plant Growth, Lytic Enzymes and Secondary Metabolites Production

Seven strains of Pseudomonas spp. were isolated from the south Gujarat region of India. Antifungal and bacterial activities of bacterial strains were evaluated against important plant pathogens in vitro, among them, PaRS was found most effective. The indole acetic acid production was recorded in all isolated Pseudomonas spp. Seed treatment with PaRS at 6 g/kg was significantly superior over other treatments for plant growth promotion (germination, shoot and root length, shoot and root weight, vigor index, and both shoot and root colonization) under standard roll towel method and pot conditions. The maximum siderophore production was observed in PaRS and medium production in PfRB, PaNS, and PfNC. PaNS and PaRS strains recorded strong HCN production but moderate production recorded in PaWP, PaWS, and PfNC. The maximum phosphate solubalization zone (22 mm) was found in PaRS. PaRS recorded maximum chitinase, ß-1,3-glucanase activity, hydrogen cyanide, and salicylic acid production as compared to other strains.

Current Perspectives on Chitinolytic Enzymes and Their Agro-Industrial Applications.

Simple SummaryChitin is a polysaccharide that forms the outer layer of many organisms, and it is widely used in industry. Chitinases are enzymes that can break down chitin into monomeric molecules and are used in the agro-industrial sectors. Because chitin is the key structural component of marine (mollusks, crustaceans, and marine invertebrates) and other species (algae, fungi, and insects), chitinases can be employed in the marine waste management and biocontrol of pathogenic fungi and harmful insects. Chitinase also has uses in the food industry, cosmetics, medicine, waste management, crop protection, and the production of single-cell proteins, among others. This study includes detailed information on the characterization, sources, and uses of chitinases in several areas.Chitinases are a large and diversified category of enzymes that break down chitin, the world’s second most prevalent polymer after cellulose. GH18 is the most studied family of chitinases, even though chitinolytic enzymes come from a variety of glycosyl hydrolase (GH) families. Most of the distinct GH families, as well as the unique structural and catalytic features of various chitinolytic enzymes, have been thoroughly explored to demonstrate their use in the development of tailor-made chitinases by protein engineering. Although chitin-degrading enzymes may be found in plants and other organisms, such as arthropods, mollusks, protozoans, and nematodes, microbial chitinases are a promising and sustainable option for industrial production. Despite this, the inducible nature, low titer, high production expenses, and susceptibility to severe environments are barriers to upscaling microbial chitinase production. The goal of this study is to address all of the elements that influence microbial fermentation for chitinase production, as well as the purifying procedures for attaining high-quality yield and purity.

Actinobacteria-enhanced plant growth, nutrient acquisition, and crop protection: Advances in soil, plant, and microbial multifactorial interactions

Agricultural areas of land are deteriorating every day owing to population increase, rapid urbanization, and industrialization. To feed today's huge populations, increased crop production is required from smaller areas, which warrants the continuous application of high doses of inorganic fertilizers to agricultural land. These cause damage to soil health and, therefore, nutrient imbalance conditions in arable soils. Under these conditions, the benefits of microbial inoculants (such as Actinobacteria) as replacements for harmful chemicals and promoting ecofriendly sustainable farming practices have been made clear through recent technological advances. There are multifunctional traits involved in the production of different types of bioactive compounds responsible for plant growth promotion, and the biocontrol of phytopathogens has reduced the use of chemical fertilizers and pesticides. There are some well-known groups of nitrogen-fixing Actinobacteria, such as Frankia, which undergo mutualism with plants and offer enhanced symbiotic trade-offs. In addition to nitrogen fixation, increasing availability of major plant nutrients in soil due to the solubilization of immobilized forms of phosphorus and potassium compounds, production of phytohormones, such as indole-3-acetic acid, indole-3-pyruvic acid, gibberellins, and cytokinins, improving organic matter decomposition by releasing cellulases, xylanase, glucanases, lipases, and proteases, and suppression of soil-borne pathogens by the production of siderophores, ammonia, hydrogen cyanide, and chitinase are important features of Actinobacteria useful for combating biotic and abiotic stresses in plants. The positive influence of Actinobacteria on soil fertility and plant health has motivated us to compile this review of important findings associated with sustaining plant productivity in the long run.

The chitinolytic activity of the Curtobacterium sp. isolated from field-grown soybean and analysis of its genome sequence.

Curtobacterium sp. GD1 was isolated from leaves of conventionally grown soybean in Brazil. It was noteworthy that among all bacteria previously isolated from the same origin, only Curtobacterium sp. GD1 showed a strong chitinase activity. The enzyme was secreted and its production was induced by the presence of colloidal chitin in the medium. The chitinase was partially purified and characterized: molecular weight was approximately 37 kDa and specific activity 90.8 U/mg. Furthermore, Curtobacterium sp. GD1 genome was sequenced and analyzed. Our isolate formed a phylogenetic cluster with four other Curtobacterium spp. strains, with ANIb/ANIm ≥ 98%, representing a new, still non described Curtobacterium species. The circular genome visualization and comparison of genome sequences of strains forming new cluster indicated that most regions within their genomes were highly conserved. The gene associated with chitinase production was identified and the distribution pattern of glycosyl hydrolases genes was assessed. Also, genes associated with catabolism of structural carbohydrates such as oligosaccharides, mixed polysaccharides, plant and animal polysaccharides, as well as genes or gene clusters associated with resistance to antibiotics, toxic compounds and auxin biosynthesis subsystem products were identified. The abundance of putative glycosyl hydrolases in the genome of Curtobacterium sp. GD1 suggests that it has the tools for the hydrolysis of different polysaccharides. Therefore, Curtobacterium sp. GD1 isolated from soybean might be a bioremediator, biocontrol agent, an elicitor of the plant defense responses or simply degrader.

Biocontrol Potential of Trichoderma afroharzianum TM24 Against Grey Mould on Tomato Plants.

Grey mould caused by Botrytis cinerea leads to severe economic loss on commercial tomato production. Application of beneficial microorganism offers an eco-friendly alternative for mitigation of tomato fungal disease damage, considering negative influences of fungicides. In the present study, an antagonistic Trichoderma afroharzianum isolate TM24 was evaluated for its biocontrol potential on tomato grey mould. The isolate TM24 showed obviously antagonistic effect on B. cinerea mycelium growth and production of glucanase and chitinase. Leaf spraying with spore suspension of isolate TM24 showed a biocontrol efficiency of over 54% against tomato grey mould in greenhouse pot experiment. The activities of plant defense-related enzymes including polyphenol oxidase, phenylalanine ammonialyase, superoxide dismutase, and peroxidase were all increased to varying degrees in tomato leaves after isolate TM24 treatment. Transcriptome analysis showed that, a total of 1941, 1753 and 38 differentially expressed genes (DEGs) were obtained at 24, 48 and 72 hpi, respectively, in tomato leaves pretreated with T. afroharzianum TM24, and then challenged with B. cinerea inoculation. The DEGs were mainly enriched in MAPK signaling pathway and plant hormones signal transduction pathway. Multiple genes that regulated crucial nodes of defense-related pathways, like flavonoid, phenylpropanoid, jasmonic acid and ethylene metabolisms were also identified, which may have positive correlations with the biocontrol potential of isolate TM24 in tomato plants. These promising results provided valuable information on using T. afroharzianum TM24 as a beneficial biocontrol agent in tomato grey mould management.

A novel Thermothelomyces heterothallicus PA2S4T fungus isolated from the soil induces chitinase production using orange peel flour

Chitinases are enzymes capable of hydrolysing the β-1,4 bonds of chitin releasing chitooligosaccharides and N-acetylglucosamine and are widely used in food, pharmaceutical, and agricultural industries. Microorganisms are potential producers of this enzyme; however, there are no reports in the literature on the production of chitinase by fungi of the genus Thermothelomyces. Thus, this work aimed to investigate the production of extracellular chitinase using alternative carbon sources by the fungus isolated from soil, Thermothelomyces heterothallicus PA2S4T. The fungus was cultivated in a liquid medium supplemented with carbon sources and incubated at 40°C under stationary conditions for seven days. Orange peel flour was the best inducer for extracellular chitinase, with 82.3 U/mL of enzymatic activity. The highest production of chitinase was obtained on the tenth day, and the optimum pH and temperature for enzyme activity were 4.5 and 50ºC, respectively. Therefore, the fungus T. heterothallicus PA2S4T proved to be promising in the production of extracellular chitinase, which presents pH and temperature characteristics favourable to biotechnological application.

Improved chromium tolerance of Medicago sativa by plant growth-promoting rhizobacteria (PGPR)

BackgroundSoil pollution by heavy metals increases the bioavailability of metals like hexavalent chromium (Cr (VI)), subsequently limiting plant growth and reducing the efficiency of phytoremediation. Plant growth-promoting rhizobacteria (PGPR) have substantial potential to enhance plant growth as well as plant tolerance to metal stress. The aim of this research was to investigate Cr (VI) phytoremediation enhancement by PGPR. ResultsThe results showed that the 27 rhizobacterial isolates studied were confirmed as Cr (VI)-resistant PGPR, by using classical biochemical tests (phosphate solubilization, nitrogen fixation, indole acetic acid, exopolysaccharides, hydrogen cyanide, siderophores, ammonia, cellulase, pectinase, and chitinase production) and showed variable levels of Cr (VI) resistance (300–600 mg/L). The best four selected Cr (VI)-resistant PGPR (NT15, NT19, NT20, and NT27) retained most of the PGP traits in the presence of 100–200 mg/L concentrations of Cr (VI). The inoculation of Medicago sativa with any of these four isolates improved the shoot and root dry weight. The NT27 isolate identified using 16S rDNA gene sequence analyses as a strain of Pseudomonas sp. was most effective in terms of plant growth promotion and stress level decrease. It increased shoot and root dry weights of M. sativa by 97.6 and 95.4%, respectively, in the presence of Cr (VI) when compared to non-inoculated control plants. It also greatly increased chlorophyll content and decreased the levels of stress markers, malondialdehyde, hydrogen peroxide, and proline. The results of the effect of Pseudomonas sp. on Cr content and bioaccumulation factor (BAF) of the shoots and roots of M. sativa plants showed the increase of plant biomass concomitantly with the increase of Cr root concentration in inoculated plants. This would lead to a higher potential of Cr (VI) phytostabilization. ConclusionsThis study demonstrates that the association M. sativa-Pseudomonas sp. may be an efficient biological system for the bioremediation of Cr (VI)-contaminated soils.

Production and Screening of Streptomyces-Extracellular Chitinase

The aim of this research was to produce Streptomyces-extracellular chitinase and screen its antifungal activity on a clinically isolated Candida albicans. The Streptomyces were isolated from an agricultural farmland; they were identified and screened for the chitinase production. Effects of time, temperature, pH and nitrogen sources on the chitinase production were determined using standard methods. Ammonium sulphate precipitation was used to partially purify the chitinase. Protein concentrations were determined spectrophotometrically using bovine serum albumin as standard. Agar-well diffusion method was used to evaluate the antifungal activity of the chitinase on C. albicans. The isolated Streptomyces were of three (3) strains, and all the strains are Gram positive, catalase positive, oxidase positive while, Strain A and C are indole positive and only Strain B is citrate positive. The maximum chitinase production was at 72 h, 40°C and when yeast extract was used as the nitrogen source. Ammonium sulphate (80%) precipitation yielded the highest enzyme activity of 39.0U/ml. The maximum enzyme activity was observed at temperature of 40oC, pH 5.5 and 1.0% colloidal chitin (substrate). The partially purified chitinase showed a zone of inhibition of 20.11 ± 1.26 mm against the Candida albicans. This result has no significant difference (P>0.05) when compared with that of the standard drug (Fluconazole) with 21.42 ± 0.08 mm zone of inhibition. These findings suggest that Streptomyces at favourable conditions produce chitinase, and this enzyme can be used as an antifungal agent on Candida albicans and other chitin containing fungi.

Screening of chitinase-producing rhizosphere actinomycetes and their genetic diversity

Abstract. Wahyudi AT, Fitriansyah NG, Amri MF, Priyanto JA, Nawangsih AA. 2021. Screening of chitinase-producing rhizosphere actinomycetes and their genetic diversity. Biodiversitas 22: 4186-4192. Soil microbial community, especially rhizosphere actinomycetes, plays an important role in protecting plants from phytopathogenic fungi by producing various agroactive compounds, including mycolytic enzymes particularly chitinase. The objective of this study was to investigate chitinase activity and analyze the diversity of family 19 chitinase genes of actinomycetes isolated from maize and soybean rhizosphere. Of fivety actinimycetes, Seventeen isolates exhibited chitinolytic activities and formed a hydrolytic zone around the colony with chitinolytic index ranging from 0.49±0.01 to 2.15±0.69, qualitatively tested in 0.3% chitin agar medium. Six selected isolates (ARJ 36, ARJ 81, ARJ 15, ARK 17, ARK 143, ARK 103) showed chitinase activities ranging from 0.157±0.04 to 0.440±0.09 U/mL, based on chitinolytic index. The production of chitinase was confirmed by the presence of family 19 chitinase encoding genes from Streptomyces sp. Some conserved regions and essential amino acid residues were also detected. This study suggests that chitinolytic actinomycetes isolated from maize and soybean rhizosphere can be studied further as biological control candidates for controlling phytopathogenic fungi.

Phylogeny and Optimization of Trichoderma harzianum for Chitinase Production: Evaluation of Their Antifungal Behaviour against the Prominent Soil Borne Phyto-Pathogens of Temperate India

Trichoderma is the most commonly used fungal biocontrol agent throughout the world. In the present study, various Trichoderma isolates were isolated from different vegetable fields. In the isolated microflora, the colony edges varied from wavy to smooth. The mycelial forms were predominantly floccose with hyaline color and conidiophores among all the strains were highly branched. Based on morphological attributes, all the isolates were identified as Trichoderma harzianum. The molecular identification using multilocus sequencing ITS, rpb2 and tef1α, genes further confirmed the morphological identification. The average chitinase activity varied from 1.13 units/mL to 3.38 units/mL among the various isolates, which increased linearly with temperature from 15 to 30 °C. There was an amplified production in the chitinase production in the presence of Mg+ and Ca2+ and Na+ metal ions, but the presence of certain ions was found to cause the down-regulated chitinase activity, i.e., Zn2+, Hg2+, Fe2+, Ag+ and K+. All the chitinase producing Trichoderma isolates inhibited the growth of tested pathogens viz., Dematophora necatrix, Fusarium solani, Fusarium oxysporum and Pythium aphanidermatum at 25% culture-free filtrate concentration under in vitro conditions. Also, under in vivo conditions, the lowest wilt incidence and highest disease control on Fusarium oxysporum was observed in isolate BT4 with mean wilt incidence and disease control of 21% and 48%, respectively. The Trichoderma harzianum identified in this study will be further used in formulation development for the management of diseases under field conditions.

Purification, characterization, and antifungal activity of Bacillus cereus strain NK91 chitinase from rhizospheric soil samples of Himachal Pradesh, India.

Newly isolated Bacillus cereus strain NK91 was characterized for extracellular chitinase production. Partially purified chitinase showed a molecular weight of 43.7 kDa in SDS-PAGE analysis. The optimum pH and temperature for the partially purified enzyme were 7.0 and 40°C, respectively. The addition of Mn2+ resulted in a 21% increase in enzyme activity as compared to the control. The Vmax and Km of the enzyme were determined as 76.9 μmol/min and 0.07 mg/mL, respectively. This enzyme exhibited stronger antifungal activity towards Fusarium oxysporum (66.7%), Rhizoctonia solani (64.6%), and Colletotrichum gloeosporioides (63%), and transmission electron microscopy and scanning transmission electron microscopy analysis showed considerable changes in cell wall structure with the treatment of purified chitinase as compared to control. Therefore, this enzyme reveals its biocontrol potential against potent phytopathogens in agriculture that can be helpful in swapping harmful as well as expensive fungicides.