scholarly journals Isolation and Identification of the Potential Novel L-asparaginase producing Bacillus Strains from Soil

2021 ◽  
pp. 11-19
Author(s):  
Naushaba Nazli ◽  
Rukhsar Masood ◽  
Muhmmad Salman ◽  
Bilal Nasir ◽  
Farah Shireen ◽  
...  

L-Asparaginase is a well know enzyme for its antineoplastic potential and is widely used to treat acute lymphoblastic leukemia and lymphosarcoma. The present work describes the isolation and characterization of novel L-asparaginase producing Bacillus strains from soil. Soil samples were collected from three different locations such as fruit garden, dairy farm and agricultural land in Peshawar Khyber Pakhtunkhwa, Pakistan. The isolates were screened to produce L-asparaginase in growth medium supplemented with 1% L-asparagine using a phenol red indicator. Among 30 bacterial isolates, only two strains initially coded as A5 and FG7 showed L-asparaginase activity. Based on biochemical and 16S rRNA sequencing analysis, the isolate A5 and FG7 were identified as Bacillus amyloliquefaciens and Bacillus proteolyticus respectively. Different factors like pH and time were optimized for maximum L-asparaginase activity. Bacillus amyloliquefaciens showed maximum asparaginase activity at pH 7 after 24 hours incubation at 30oC, while Bacillus proteolyticus showed optimum activity at pH 7 after 48 hours of incubation at 30oC. The present study first time reported the production of L-asparginase enzyme from Bacillus amyloliquefaciens and Bacillus proteolyticus. Keywords: L-asparaginase, Bacillusamyloliquefaciens, Bacillus proteolyticus, 16sRNA.

2020 ◽  
Vol 2 (2) ◽  
pp. 46-55 ◽  
Author(s):  
Gholamhossein Ebrahimipour ◽  
Mohammad Yaghoobi Avini ◽  
Mahtab Ghorbanmovahhed

L-asparaginase is an important therapeutic enzyme used in combination with other drugs for therapy of Acute Lymphoblastic Leukemia (ALL). L-asparaginase catalyzes the conversion of asparagine to aspartic acid and ammonia. In recent years, this enzyme gained applications in many fields of science such as clinical research, pharmacological, and food industries. This study was aimed at isolation and identification of a strain with the ability to producing extracellular glutaminase free L-asparaginase from soil and determination of enzyme stability. The isolation was done on M9 medium. Biochemical tests and 16S rDNA sequence was used for strain identification. L-asparaginase was partially purified using ammonium sulfate precipitation, dialysis, and DEAE-anion exchange chromatography. The effect of pH and temperature on enzyme activity was investigated. The isolated bacteria were identified as Staphylococcus sp. The optimum pH and temperature for maximum L-asparaginase activity were found at 8 and 35 °C. The enzyme purification showed a single band around 115 kDa on SDS-Page. The optimal activity for the enzyme produced by MGM1 was similar to the physiological conditions of the human body, therefore, further studies on this enzyme would be of great value in finding a new efficient asparaginase enzyme.


2021 ◽  
Vol 11 (10) ◽  
pp. 4466
Author(s):  
Hyeji Lim ◽  
Sujin Oh ◽  
Sungryul Yu ◽  
Misook Kim

The purpose of this study was to isolate functional Bacillus strains from Korean fermented soybeans and to evaluate their potential as probiotics. The L-asparaginase activity of MKHJ 1-1 was the highest among 162 Bacillus strains. This strain showed nonhemolysis and did not produce β-glucuronidase. Among the nine target bacteria, MKHJ 1-1 inhibited the growth of Escherichia coli, Pseudomonas aeruginosa, Shigella sonnei, Shigella flexneri, Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus cereus. 16S rRNA gene sequence analysis resulted in MKHJ 1-1 identified as Bacillus subtilis subsp. stercoris D7XPN1. As a result of measuring the survival rate in 0.1% pepsin solution (pH 2.5) and 0.3% bile salt solution for 3 h, MKHJ 1-1 exhibited high acid resistance and was able to grow in the presence of bile salt. MKHJ 1-1 showed outstanding autoaggregation ability after 24 h. In addition, its coaggregation with pathogens was strong. Therefore, MKHJ 1-1 is a potential probiotic with L-asparaginase activity and without L-glutaminase activity, suggesting that it could be a new resource for use in the food and pharmaceutical industry.


1995 ◽  
Vol 60 (9) ◽  
pp. 1555-1572 ◽  
Author(s):  
Pavel Kramata ◽  
Jaroslav Černý ◽  
Gabriel Birkuš ◽  
Ivan Votruba ◽  
Berta Otová ◽  
...  

Using a single isolation procedure and selective assays for the determination of enzyme activity, highly purified DNA-polymerases α, δ and ε were isolated from the lymphoma of Sprague-Dawley inbred rats. For pol α the subunit composition was 170, 70, 57 and 53 kDa with sedimentation coefficient 8.7 S for the native molecule; pol delta consists of two polypeptides (133 and 46 kDa; sedimentation coefficient 8.2 S), while pol ε is a single polypeptide (140 kDa) and its sedimentation coefficient is 7.0 S. Comparison of the interaction of individual enzymes with known inhibitors and proliferating cell nuclear antigen (PCNA) using the template-primer poly dA-oligo dT12-18, gave the following data: (i) pol α is selectively inhibited by N2-(p-butylphenyl)-2'-deoxyguanosine 5'-triphosphate (BuPdGTP) and stimulated by dimethyl sulfoxide; (ii) all the enzymes are inhibited by N-ethylmaleimide and aphidicolin; (iii) PCNA stimulates pol δ approximately 50 times while pol ε is moderately inhibited; (iv) pol α exhibits considerably higher DNA-primase activity with poly dC as template than with poly dT, and negligible 3'-5'-exonuclease activity whereas pol δ and pol ε, which do not exert any DNA-primase activity have approximately the same 3'-5'-exonuclease activity. The ability of individual polymerases to utilize poly dT-oligo dA12-18 as a template-primer at different pH values, ionic strengths and Mg2+-concentrations was also investigated. In comparison to poly dA-oligo dT12-18 template-primer, pol α has 140% of enzyme activity on poly dT-oligo dA12-18 under optimal conditions, whereas the activity of pol ε and pol δ is 4 times and 10 times lower, respectively.


Author(s):  
Harjeet Singh ◽  
Shweta Sao

L-asparaginase (EC 3.5.11. L-asparagine amidohydrolase) is first enzyme, studied very intensively in human beings with regard to its anti-tumor potential against tumor of lymphoid precursor, acute lymphoblastic leukemia (ALL). The current drugs are suffering from many side effects like immune suppression, infertility, secondary neoplasm. The immunogenic complications associated with its present microbial sources Escherichia coli; Erwinia carotovora limits its medicinal frontier. So there exists a need of switching to novel natural sources to serve as non-immunogenic and better production sources of L-asparaginase. In the present study, four cultures of fungal endophytes viz. TSF-1, TSF-2, TSF-3 and TSF-4 selected on the basis of primary and secondary screening was carried on with L-asparagine as a sole carbon and nitrogen source and phenol red as pH indicator. The maximum protein content was observed to be present in TSF-2 i.e. 2.727 mg /mL and possessed maximum activity of 6.054 Units/ml. Sample was separated by SDS-PAGE, stained by silver staining, showed a single band with molecular weight of approximately ~45kDa.


2021 ◽  
Vol 10 (Supplement_2) ◽  
pp. S19-S19
Author(s):  
Johanny Contreras ◽  
Karina Rivera ◽  
María Castillo ◽  
Genara Santana ◽  
María Dolores Gil ◽  
...  

Abstract Background In October 2018, the Hispaniola Project was initiated to build local expertise in infection care and prevention at three pediatric oncology units (POUs) in Haiti and the Dominican Republic. Surveillance of healthcare-associated infections (HAI) was a central aim. Severe and prolonged neutropenia is a frequent risk factor for infections in oncology patients. Among HAIs, bacteremia is one of the most serious; bacteremia requires timely isolation and identification of the offending microorganism and the antimicrobial susceptibility. These diagnostic interventions allow informed therapeutic and prophylactic measures. Here, we report our experience in bacteremia in these 3 POUs. Methods We conducted prospective infection surveillance of all patients admitted to three POUs in Hispaniola Island. Blood culture methods followed standard national procedures. We used the 2018 US Centers for Disease Control National Healthcare Safety Network case definitions for primary laboratory-confirmed bloodstream infections (LCBI), and we categorized infections as healthcare-associated or present on admission (POA). We reviewed data collected from January 2019 to December 2020 and used descriptive statistics to report our results. Results Our review identified 66 LCBIs with an overall rate of 3.52 infections per 1000 patient-days. Of these, 40 (61%) were healthcare-associated, and 26 were POA. The majority (41, 62%) of patients were undergoing chemotherapy at the time of the infection, with induction being the most common phase (23). The most common oncologic diagnosis was acute lymphoblastic leukemia (43, 65%), followed by solid tumor (12, 18%). Fifty-three (80%) of the infections met the LCBI-1 criteria, with the other 13 categorized as LBCI-2. Of the 53 LCBI-1, 7 (13%) were considered related to mucosal barrier injury (MBI-LCBI 1 definition). The most commonly identified organisms were Klebsiella spp. (13, 19%) and coagulase-negative Staphylococcus (13, 19%). Antibiotic resistance was observed in many of the identified pathogens, with nearly half (25, 44%) of the 57 bacterial isolates having any resistance and a quarter (14, 25%) with resistance to multiple classes, including cephalosporins, fluoroquinolones, and aminoglycosides. Eleven (17%) patients were admitted to the Intensive Care Unit as a result of the LCBI. Thirteen deaths were recorded among the patients with LCBIs, with 6 (46%) associated with the HAI and 7 (54%) related to disease progression. Conclusions Our findings demonstrate that resistant pathogens were frequent among the LCBI isolates. Our preliminary results are guiding clinical management to be vigilant in our care of patients at high risk for bacteremia and poor clinical response by initiating more effective antimicrobials sooner. Importantly, reviewing reasons for antimicrobial resistance and implementing best antimicrobial use practices will protect our fragile antibiotic arsenal. Infection surveillance programs, such as ours, and other initiatives which promote infection prevention and control in POU will increase the quality of care for these vulnerable patients.


2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Gianvito Lanave ◽  
Vittorio Larocca ◽  
Michele Losurdo ◽  
Cristiana Catella ◽  
Paolo Capozza ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document