L-Asparaginase (L-ASNase) is a crucial anti-tumour drug used to cure acute lymphocytic leukaemia. The current study aimed to enhance the production medium for the endophytic fungus Curvularia sp. LCJ413 that showed significant L-ASNase activity. L-ASNase production from Curvularia sp. LCJ413 was examined in six different media to select an appropriate liquid medium. Among the various media tested, Modified Czapek Dox broth (MCDB) exhibited the maximum L-ASNase activity (8.81 ± 0.52 U/mL). Physical (pH and temperature) and nutritional (carbon, nitrogen, inducer, and their concentrations) parameters were also optimized to boost L-ASNase production. Results of the study suggested a temperature of 28°C at pH 7 with 2 g/L maltose, 10 g/L L-Asparagine, and 25 g/L ammonium sulphate as the optimal carbon, inducer, and nitrogen source resulted in a high L-ASNase activity of 18.9 ± 0.40 U/mL. The statistical enhancement of L-ASNase by Response Surface Methodology (RSM) produced 20.11 U/mL of L-ASNase, which was 2.2 fold higher than the non-optimized medium. This is the first study on L-ASNase production from the endophytic Curvularia sp. LCJ413 isolated from Vitex negundo medicinal plant. Continuous fermentation with the medium composition provided in the study can produce L-ASNase on a large scale.
Antibiotic resistance, an emerging threat to human health, is the world's major challenge today. Endophytic microbes found in medicinal plants aid in the resolution of these issues. It is critical to select the plant that will be used to investigate endophytes for bioactive metabolites. As a result, medicinal plants which have been used as a traditional source of medicine for centuries, are a valuable source for bioprospecting endophytes. Fungi are the most commonly reported organisms among endophytes. Endophytic fungi are a diverse group of microbes that invade plant interior tissues without creating any symptoms. Endophytic fungi represent an abundant source of novel bioactive secondary metabolites with diversified structures that can be exploited for various applications in the field of therapeutics. Endophytic fungi produce secondary metabolites such as alkaloids, enzymes, steroids, terpenoids, phenols, quinones, peptides, polyketides etc. which have a higher therapeutic value. They produce a wide range of bioactive molecules with various biological activities such as antibacterial, antiviral, antifungal, antioxidants, cytotoxic and antimalarial compounds. The chemical diversity of endophytic fungi also serves to protect host plants from pathogens by hindering their growth and stimulating the host immune system, thereby enhancing plant defense mechanisms. Endophytic fungi synthesize metabolites that are identical to their host medicinal plant (azadirachtin, taxol, ginkgolide, jasmonic acid, camptothecin etc.) due to their specific capacity to form a symbiotic association with the host. Endophytic fungi can serve as substitutes for secondary metabolite production, aiding in biodiversity conservation. This review discusses numerous bioactive metabolites produced by endophytic fungi that have diverse biological activities. The majority of these metabolites have the potential to be utilized as drugs to treat a wide range of diseases.
Endophytic fungi synthesize a diverse set of enzymes and biomolecules with wide array of bioactivities. Thirty-five distinct fungal endophytes have been isolated from medicinal plants of Rutaceae and screened for their ability to produce various industrially important enzymes. Primary enzyme screening revealed that 65% of isolates produced L-Asparaginase, 62% of isolates produced cellulase, 60% of isolates produced amylase, 48% of isolates produced both lipase and pectinase, 45% of isolates produced laccase and 42% of isolates produced protease. Among the different enzymes screened, L-Asparaginase producing isolates were dominant and subjected to quantification using the nesslerization method. The endophytic isolates LCJ315, LCJ324, LCJ326 and LCJ335 were capable of producing high L-Asparaginase activity and their enzyme activity ranged between 7.58 U/ml to 8.84 U/ml. The four isolates were further subjected to molecular identification using 18S rRNA sequencing. The results were then subjected to BLAST analysis. The phylogenetic tree was created by MEGA software. The isolated endophytic fungal strains were identified as LCJ315 (Aspergillus sp.), LCJ326 (Colletotrichum sp.), LCJ324 and LCJ335 (Fusarium sp.). The fungal source of L-Asparaginase is preferred over other microbial sources of L-Asparaginase due to reduced side effects. Given the fact that L-Asparaginase is a crucial anticancer enzyme that is used as the initial therapy for acute lymphoblastic leukemia. Hence, the present study reveals that these endophytic fungal strains isolated from Rutaceae members can be used as an alternate source for L-Asparaginase production.
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