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Journal of Microbiological Methods Dec 2021L-asparaginase from endophytic Fusarium proliferatum (isolate CCH, GenBank accession no. MK685139) isolated from the medicinal plant Cymbopogon citratus (Lemon grass),...
L-asparaginase from endophytic Fusarium proliferatum (isolate CCH, GenBank accession no. MK685139) isolated from the medicinal plant Cymbopogon citratus (Lemon grass), was optimized for its L-asparaginase production and its subsequent cytotoxicity towards Jurkat E6 cell line. The following factors were optimized; carbon source and concentration, nitrogen source and concentration, incubation period, temperature, pH and agitation rate. Optimization of L-asparaginase production was performed using One-Factor-At-A-Time (OFAT) and Response surface methodology (RSM) model. The cytotoxicity of the crude enzyme from isolate CCH was tested on leukemic Jurkat E6 cell line. The optimization exercise revealed that glucose concentration, nitrogen source, L-asparagine concentration and temperature influenced the L-asparaginase production of CCH. The optimum condition suggested using OFAT and RSM results were consistent. As such, the recommended conditions were 0.20% of glucose, 0.99% of L-asparagine and 5.34 days incubation at 30.50 °C. The L-asparaginase production of CCH increased from 16.75 ± 0.76 IU/mL to 22.42 ± 0.20 IU/mL after optimization. The cytotoxicity of the crude enzyme on leukemic Jurkat cell line recorded IC value at 33.89 ± 2.63% v/v. To conclude, the enzyme extract produced from Fusarium proliferatum under optimized conditions is a potential alternative resource for L-asparaginase.
Topics: Antineoplastic Agents; Asparaginase; Carbon; Culture Media; Cytotoxins; Databases, Nucleic Acid; Endophytes; Fusarium; Hydrogen-Ion Concentration; Microbiological Techniques; Nitrogen; Plants, Medicinal; Temperature
PubMed: 34743930
DOI: 10.1016/j.mimet.2021.106358 -
Leukemia & Lymphoma Apr 2023The addition of asparaginase to acute lymphoblastic leukemia (ALL) and lymphoblastic lymphoma (LBL) treatment regimens provides significant patient benefits....
The addition of asparaginase to acute lymphoblastic leukemia (ALL) and lymphoblastic lymphoma (LBL) treatment regimens provides significant patient benefits. Asparaginase therapies vary in origin ( or -derived) and preparation (native or pegylated), conferring distinct pharmacokinetic and immunogenic profiles. Clinical hypersensitivity reactions (HSRs) are commonly reported in patients and range from localized erythema to systemic anaphylaxis. Due to its favorable pharmacokinetic profile and reduced immunogenicity compared to native preparations, pegaspargase is the first-line asparaginase therapeutic option. Switching to an -derived asparaginase is recommended for patients who experience HSRs or antibody-mediated inactivation to achieve the significant clinical benefit observed in patients who complete asparaginase treatment. Previous global shortages of asparaginase necessitated conversion mitigation strategies such as premedication protocols, desensitization, and asparaginase activity level monitoring. Here, we discuss the efficacy, safety, pharmacokinetics, current use, and administration of asparaginase therapies for pediatric and adolescent patients with ALL/LBL.
Topics: Adolescent; Humans; Child; Asparaginase; Escherichia coli; Expert Testimony; Drug Hypersensitivity; Dickeya chrysanthemi; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Hypersensitivity; Antineoplastic Agents
PubMed: 36781296
DOI: 10.1080/10428194.2023.2171267 -
Biochemistry Sep 2020l-Asparaginase (EC 3.5.1.1) was first used as a component of combination drug therapies to treat acute lymphoblastic leukemia (ALL), a cancer of the blood and bone...
l-Asparaginase (EC 3.5.1.1) was first used as a component of combination drug therapies to treat acute lymphoblastic leukemia (ALL), a cancer of the blood and bone marrow, almost 50 years ago. Administering this enzyme to reduce asparagine levels in the blood is a cornerstone of modern clinical protocols for ALL; indeed, this remains the only successful example of a therapy targeted against a specific metabolic weakness in any form of cancer. Three problems, however, constrain the clinical use of l-asparaginase. First, a type II bacterial variant of l-asparaginase is administered to patients, the majority of whom are children, which produces an immune response thereby limiting the time over which the enzyme can be tolerated. Second, l-asparaginase is subject to proteolytic degradation in the blood. Third, toxic side effects are observed, which may be correlated with the l-glutaminase activity of the enzyme. This Perspective will outline how asparagine depletion negatively impacts the growth of leukemic blasts, discuss the structure and mechanism of l-asparaginase, and briefly describe the clinical use of chemically modified forms of clinically useful l-asparaginases, such as Asparlas, which was recently given FDA approval for use in children (babies to young adults) as part of multidrug treatments for ALL. Finally, we review ongoing efforts to engineer l-asparaginase variants with improved therapeutic properties and briefly detail emerging, alternate strategies for the treatment of forms of ALL that are resistant to asparagine depletion.
Topics: Adolescent; Asparaginase; Asparagine; Child; Child, Preschool; Glutaminase; Humans; Medical Oncology; Models, Molecular; Polyethylene Glycols; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Protein Conformation; Quality Improvement; Young Adult
PubMed: 32786406
DOI: 10.1021/acs.biochem.0c00354 -
Pharmacogenomics May 2022To investigate the role of gene and rs3809849 in pancreatic cancer (PANC1) and lymphoblastic leukemia (NALM6) cell lines and their response to asparaginase treatment....
To investigate the role of gene and rs3809849 in pancreatic cancer (PANC1) and lymphoblastic leukemia (NALM6) cell lines and their response to asparaginase treatment. The authors applied CRISPR-Cas9 to produce knock-out (KO) and rs3809849 knock-in (KI) cell lines. The authors also interrogated rs3809849's impact on PANC1 cells through allele-specific overexpression. PANC1 KO cells exhibited lower proliferation capacity (p ≤ 0.05), higher asparaginase sensitivity (p = 0.01), reduced colony-forming potential (p = 0.001), cell cycle blockage in S phase, induction of apoptosis and remarkable morphology changes suggestive of an epithelial-mesenchymal transition. Overexpression of the wild-type (but not the mutant) allele of -rs3809849 in PANC1 cells increased asparaginase sensitivity. NALM6 KO displayed resistance to asparaginase (p < 0.0001), whereas no effect for rs3809849 KI was noted. is important for regulating various cellular functions, and it plays, along with its rs3809849 polymorphism, a tissue-specific role in asparaginase treatment response.
Topics: Alleles; Asparaginase; DNA-Binding Proteins; Humans; Pancreatic Neoplasms; Precursor Cell Lymphoblastic Leukemia-Lymphoma; RNA-Binding Proteins; Transcription Factors
PubMed: 35485735
DOI: 10.2217/pgs-2022-0010 -
Future Microbiology Jan 2024To review the available literature about heterologous expression of fungal L-asparaginase (L-ASNase). A search was conducted across PubMed, Science Direct, Scopus and... (Review)
Review
To review the available literature about heterologous expression of fungal L-asparaginase (L-ASNase). A search was conducted across PubMed, Science Direct, Scopus and Web of Science databases; 4172 citations were identified and seven articles were selected. The results showed that heterologous expression of fungal L-ASNase was performed mostly in bacterial expression systems, except for a study that expressed L-ASNase in a yeast system. Only three publications reported the purification and characterization of the enzyme. The information reported in this systematic review can contribute significantly to the recognition of the importance of biotechnological techniques for L-ASNase production.
Topics: Humans; Asparaginase; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Bacteria; Antineoplastic Agents
PubMed: 37882841
DOI: 10.2217/fmb-2023-0131 -
Allergologia Et Immunopathologia 2023L-Asparaginase (L-asp), the unconjugated form of polyethylene glycol-conjugated L-asparaginase (PEG-asp), regulates T cell stimulation, antibody production, and...
BACKGROUND
L-Asparaginase (L-asp), the unconjugated form of polyethylene glycol-conjugated L-asparaginase (PEG-asp), regulates T cell stimulation, antibody production, and lysosomal protease activity to mediate PEG-asp-related anaphylaxis. This study aimed to investigate the relation of L-asp activity and anti-L-asp antibody with anaphylaxis risk and non-anaphylaxis adverse reaction risk in childhood acute lymphoblastic leukemia (ALL) patients who underwent PEG-asp contained therapy.
METHODS
In total, 170 childhood ALL patients underwent PEG-asp-contained treatment and their L-asp activity and anti-L-asp antibody were detected on the 7 day after treatment initiation.
RESULTS
There were 27 (15.9%) patients who had PEG-asp-related adverse reaction: 17 (10.0%) patients experienced PEG-asp-related anaphylaxis and 14 (8.2%) patients experienced PEG- asp-related non-anaphylaxis adverse reaction. Moreover, L-asp activity was negatively related to anti-L-asp antibody in childhood ALL patients (<0.001). Elevated L-asp activity was associated with the absence of PEG-asp-related anaphylaxis (<0.001), PEG-asp-related non-anaphylaxis adverse reaction (=0.004), and PEG-asp-related adverse reaction (<0.001). However, the anti- L-asp antibody displayed opposite trend similar to L-asp activity. Receiver operating characteristic (ROC) curve analyses exhibited L-asp activity and anti-L-asp antibody exhibited superior predictive values in estimating PEG-asp-related anaphylaxis risk with area under curve (AUC) of 0.955 and 0.905, respectively compared to PEG-asp-related non-anaphylaxis adverse reaction risk with AUC of 0.730 and 0.675, respectively. Besides, patients with de novo disease, higher risk stratification, and allergic history showed trends linked with PEG-asp-related anaphylaxis risk.
CONCLUSION
The monitoring of L-asp activity and anti-L-asp antibody maybe useful for early estimation and prevention of PEG-asp-related anaphylaxis in childhood ALL management.
Topics: Humans; Anaphylaxis; Asparaginase; Biomarkers; Polyethylene Glycols; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Child
PubMed: 37169557
DOI: 10.15586/aei.v51i3.771 -
Microbial production, molecular modification, and practical application of l-Asparaginase: A review.International Journal of Biological... Sep 2021L-Asparaginase (L-ASNase, EC 3.5.1.1), an antitumor drug for acute lymphoblastic leukemia (ALL) therapy, is widely used in the clinical field. Similarly, L-ASNase is... (Review)
Review
L-Asparaginase (L-ASNase, EC 3.5.1.1), an antitumor drug for acute lymphoblastic leukemia (ALL) therapy, is widely used in the clinical field. Similarly, L-ASNase is also a powerful and significant biological tool in the food industry to inhibit acrylamide (AA) formation. This review comprehensively summarizes the latest achievements and improvements in the production, modification, and application of microbial L-ASNase. To date, the expression levels and optimization of expression hosts such as Escherichia coli, Bacillus subtilis, and Pichia pastoris, have made significant progress. In addition, examples of successful modification of L-ASNase such as decreasing glutaminase activity, increasing the in vivo stability, and enhancing thermostability have been presented. Impressively, the application of L-ASNase as a food addition aid, as well as its commercialization in the pharmaceutical field, and cutting-edge biosensor application developments have been summarized. The presented results and proposed ideas could be a good guide for other L-ASNase researchers in both scientific and practical fields.
Topics: Antineoplastic Agents; Asparaginase; Bacillus subtilis; Bacterial Proteins; Enzyme Stability; Escherichia coli; Food Handling; Fungal Proteins; Industrial Microbiology; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Protein Conformation; Protein Denaturation; Saccharomycetales; Structure-Activity Relationship; Substrate Specificity; Temperature
PubMed: 34293360
DOI: 10.1016/j.ijbiomac.2021.07.107 -
Applied Microbiology and Biotechnology May 2022L-asparaginase catalyzes the hydrolysis of L-asparagine to L-aspartic acid and ammonia. It has application in the treatment of acute lymphoblastic leukemia in children,...
L-asparaginase catalyzes the hydrolysis of L-asparagine to L-aspartic acid and ammonia. It has application in the treatment of acute lymphoblastic leukemia in children, as well as in other malignancies, in addition to its role as a food processing aid for the mitigation of acrylamide formation in the baking industry. Its use in cancer chemotherapy is limited due to problems such as its intrinsic glutaminase activity and associated side effects, leading to an increased interest in the search for novel L-asparaginases without L-glutaminase activity. This study reports the cloning and expression of an L-asparaginase contig obtained from whole metagenome shotgun sequencing of Sardinella longiceps gut microbiota. Purified recombinant glutaminase-free L-asparaginase SlpA was a 74 kDa homodimer, with maximal activity at pH 8 and 30 °C. K and V of SlpA were determined to be 3.008 mM and 0.014 mM/min, respectively. SlpA displayed cytotoxic activity against K-562 (chronic myeloid leukemia) and MCF-7 (breast cancer) cell lines with IC values of 0.3443 and 2.692 U/mL, respectively. SlpA did not show any cytotoxic activity against normal lymphocytes and was proved to be hemocompatible. Pre-treatment of biscuit and bread dough with different concentrations of SlpA resulted in a clear, dose-dependent reduction of acrylamide formation during baking. KEY POINTS: • Cloned and expressed L-asparaginase (SlpA) from fish gut microbiota • Purified SlpA displayed good cytotoxicity against K-562 and MCF-7 cell lines • SlpA addition caused a significant reduction of acrylamide formation during baking.
Topics: Acrylamide; Animals; Antineoplastic Agents; Asparaginase; Asparagine; Gastrointestinal Microbiome; Glutaminase
PubMed: 35579684
DOI: 10.1007/s00253-022-11954-7 -
Pediatric Blood & Cancer Jan 2020Glucocorticoids and asparaginase, used to treat acute lymphoblastic leukemia (ALL), can cause hypertriglyceridemia. We compared triglyceride levels, risk factors, and...
BACKGROUND
Glucocorticoids and asparaginase, used to treat acute lymphoblastic leukemia (ALL), can cause hypertriglyceridemia. We compared triglyceride levels, risk factors, and associated toxicities in two ALL trials at St. Jude Children's Research Hospital with identical glucocorticoid regimens, but different asparaginase formulations. In Total XV (TXV), native Escherichia coli l-asparaginase was front-line therapy versus the pegylated formulation (PEG-asparaginase) in Total XVI (TXVI).
PROCEDURE
Patients enrolled on TXV (n = 498) and TXVI (n = 598) were assigned to low-risk (LR) or standard/high-risk (SHR) treatment arms (ClinicalTrials.gov identifiers: NCT00137111 and NCT00549848). Triglycerides were measured four times and were evaluable in 925 patients (TXV: n = 362; TXVI: n = 563). The genetic contribution was assessed using a triglyceride polygenic risk score (triglyceride-PRS). Osteonecrosis, thrombosis, and pancreatitis were prospectively graded.
RESULTS
The largest increase in triglycerides occurred in TXVI SHR patients treated with dexamethasone and PEG-asparaginase (4.5-fold increase; P <1 × 10 ). SHR patients treated with PEG-asparaginase (TXVI) had more severe hypertriglyceridemia (>1000 mg/dL) compared to native l-asparaginase (TXV): 10.5% versus 5.5%, respectively (P = .007). At week 7, triglycerides did not increase with dexamethasone treatment alone (LR patients) but did increase with dexamethasone plus asparaginase (SHR patients). The variability in triglycerides explained by the triglyceride-PRS was highest at baseline and declined with therapy. Hypertriglyceridemia was associated with osteonecrosis (P = .0006) and thrombosis (P = .005), but not pancreatitis (P = .4).
CONCLUSION
Triglycerides were affected more by PEG-asparaginase than native l-asparaginase, by asparaginase more than dexamethasone, and by drug effects more than genetics. It is not clear whether triglycerides contribute to thrombosis and osteonecrosis or are biomarkers of the toxicities.
Topics: Adolescent; Asparaginase; Child; Child, Preschool; Drug Compounding; Female; Follow-Up Studies; Humans; Hypertriglyceridemia; Infant; Male; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Prognosis
PubMed: 31612640
DOI: 10.1002/pbc.28040 -
Recent Patents on Biotechnology 2019L-ASNase (L-asparagine aminohydrolase EC 3.5.1.1) is used for the conversion of L-asparagine to L-aspartic acid and ammonia and also it was found as an agent of... (Review)
Review
BACKGROUND
L-ASNase (L-asparagine aminohydrolase EC 3.5.1.1) is used for the conversion of L-asparagine to L-aspartic acid and ammonia and also it was found as an agent of chemotherapeutic property according to recent patents. It is known as an anti-cancer agent and recently it has received an immense attention. Various microorganisms have the ability to secrete the L-ASNase. It is famous world-wide as anti-tumor medicine for acute lymphoblastic leukemia and lymphosarcoma. L-ASNase helps in deamination of Asparagine and Glutamine.
SOURCE
L-ASNase mainly found in two bacterial sources; Escherichia coli and Erwinia carotovora. Isolation from plants: Endophytes were also a great source of L-ASNase. It was isolated from four types of plants named as; C. citratus, O. diffusa, M. koengii, and also P. bleo.
APPLICATIONS
L-ASNase is used as a potential anti-tumor medicine. It plays a very much essential role for the growth of tumor cells. Tumor cells require a lot of asparagine for their growth. But ASNase converts to aspartate and ammonia from asparagine. So the tumor cell does not proliferate and fails to survive. The L-ASNase is used as the medicine for the major type of cancer like acute lymphocytic leukemia (ALL), brain. It also used as a medicine for central nervous system (CNS) tumors, and also for neuroblastoma. Two types of L-ASNase have been found.
CONCLUSION
L-ASNase becomes a powerful anti-tumor medicine and researchers should develop a potent strain of asparaginase which can produce asparaginase in the industrial level. It is also used in the pharmaceutical industry and food industry on a broader scale.
Topics: Antineoplastic Agents; Asparaginase; Asparagine; Escherichia coli; Glutamine; Humans; Neuroblastoma; Patents as Topic; Pectobacterium carotovorum
PubMed: 30318009
DOI: 10.2174/1872208312666181012150407