Bioprocess Optimization for Enhanced Production of L-asparaginase via Two Model-Based Experimental Designs by Alkaliphilic Streptomyces fradiae NEAE-82

2020 ◽  
Vol 9 (1) ◽  
pp. 23-37
Author(s):  
Hoda M. Soliman ◽  
Noura El-A. El-Naggar ◽  
Sara M. El-Ewasy
Keyword(s):  
Experimental Design ◽  
Lymphoblastic Leukemia ◽  
Basal Medium ◽  
Fold Increase ◽  
Chemotherapeutic Agents ◽  
Inoculum Size ◽  
P Value ◽  
Streptomyces Fradiae ◽  
Bioprocess Optimization ◽  
Enhanced Production

Background: L-asparaginase is one of the most widely used chemotherapeutic agents for the treatment of a variety of lymphoproliferative disorders and particularly acute lymphoblastic leukemia. Due to increased applications of L-asparaginase in several industrial fields including food processing and medical fields, its production needs to be increased to several folds. Objective: The aim was (i) to identify the significant factors which affect L-asparaginase production by Streptomyces fradiae NEAE-82 and (ii) to achieve higher production of L-asparaginase. Methods: Sixteen assigned factors and three dummy factors were screened using Plackett-Burman experimental design to determine the most important factors for the production of L-asparaginase by Streptomyces fradiae NEAE-82. Results: L-asparagine was determined to be the most significant positive independent factor (P-value 0.0092) affecting L-asparaginase production by Streptomyces fradiae NEAE-82 followed by pH and NaCl with significant P-values of 0.0133 and 0.0272; respectively. These factors were further optimized by Box-Behnken experimental design. The optimized fermentation conditions, which resulted in the maximum L-asparagine activity of 53.572 UmL-1 are g/L: dextrose 4, L-asparagine 15, KNO3 2, MgSO4.7H2O 0.5, K2HPO4 1, FeSO4.7H2O 0.02, NaCl 0.2, ZnSO4 0.01 and inoculum size 2 %, v/v for 7 days incubation at temperature 37°C, agitation speed 100 rpm, pH 8.5. Conclusion: A total of 3.41-fold increase in the production of L-asparaginase was achieved in the medium after statistical improvement (53.572 UmL-1) as compared to the unoptimized basal medium used prior to the application of Plackett-Burman (15.704 UmL-1).

scholarly journals Optimization of Protease Production fromAspergillus OryzaeSp. Using Box-Behnken Experimental Design

2007 ◽  
Vol 4 (2) ◽  
pp. 145-153 ◽  
Author(s):  
G. Srinu Babu ◽  
R. R. Shiva Kiran ◽  
N. Lokeswari ◽  
K. Jaya Raju
Keyword(s):  
Response Surface Methodology ◽  
Enzyme Activity ◽  
Experimental Design ◽  
Empirical Model ◽  
Experimental Verification ◽  
Shake Flask ◽  
Basal Medium ◽  
Fold Increase ◽  
Protease Production ◽  
The Relationship

Protease production byAspergillus oryzaewas optimized in shake-flask cultures using Box-Behnken experimental design. An empirical model was developed through response surface methodology to describe the relationship between tested variable (peptone, glucose, soyabeanmeal and pH). Maximum enzyme activity was attained with Peptone at 4 g∕L; temperature at 30 °C glucose at 6 g∕L; 30 °C and pH at 10. Experimental verification of the model showed a validation of 95%, which is more than 3-fold increase compare to the basal medium.

scholarly journals Optimization of Culture Conditions for Production of the Anti-Leukemic Glutaminase Free L-Asparaginase by Newly IsolatedStreptomyces olivaceusNEAE-119 Using Response Surface Methodology

2015 ◽  
Vol 2015 ◽  
pp. 1-17 ◽  
Author(s):  
Noura El-Ahmady El-Naggar ◽  
Hassan Moawad ◽  
Nancy M. El-Shweihy ◽  
Sara M. El-Ewasy
Keyword(s):  
Response Surface Methodology ◽  
Experimental Design ◽  
Response Surface ◽  
Lymphoblastic Leukemia ◽  
Biochemical Properties ◽  
Inoculum Size ◽  
Agitation Speed ◽  
16S Rrna Sequence ◽  
Inoculum Age ◽  
Streptomyces Olivaceus

Among the antitumor drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological, and biochemical properties together with 16S rRNA sequence asStreptomyces olivaceusNEAE-119 and sequencing product (1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production byStreptomyces olivaceusNEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out. Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

scholarly journals ORGANOGENESIS FROM CEPHALOTAXUS HARRINGTONIA CALLUS: ESTABLISHMENT OF SHOOTS AND ROOT CULTURES

HortScience ◽  
1991 ◽  
Vol 26 (5) ◽  
pp. 479c-479 ◽  
Author(s):  
Enaksha R. Wickremesinhe ◽  
Richard N. Arteca
Keyword(s):  
Basal Medium ◽  
Fold Increase ◽  
Chemotherapeutic Agents ◽  
Light Regime ◽  
Growth Characteristics ◽  
Root Tip ◽  
Stem Explants ◽  
Root Cultures ◽  
Fresh Weight ◽  
Basal Salt Medium

Fast growing callus was derived from Cephalotaxus harringtonia stem explants placed on MS basal salt medium with B-5 vitamins, 3% sucrose, 1 μM kinetin and 4.5 μM 2,4-D. Callus placed on basal medium with 10 μM kinetin and 0.45 μM 2,4-D turned green and organogenesis was observed upon subculture onto basal medium without hormones. Shoots were excised and placed on 1/2 strength MS salts and 10% sucrose for further shoot development. During the process of organogenesis, we also observed the differentiation of roots. Rapidly growing root cultures were established by culturing them under a 24 hour light regime of 35 μM/m2/s. Two grams of root tip explants cultured on B-5 medium with 2% sucrose were capable of producing an average of 24 grams of roots within 11 days. A 20-fold increase in fresh weight was achieved within 3 weeks when 15 grams of these roots were cultured in a 3 liter air-sparged bioreactor. C. harringtonia contains a number of alkaloids that exhibit cytotoxicity and are being evaluated as chemotherapeutic agents. We are currently in the process of establishing growth characteristics for these roots and assaying roots for the presence of these alkaloids. All cultures were grown under a 12 hour light regime unless otherwise stated.

Activation of cytochrome c to a peroxidase compound I-type intermediate by H2O2: relevance to redox signalling in apoptosis

2004 ◽  
Vol 71 ◽  
pp. 97-106 ◽  
Author(s):  
Mark Burkitt ◽  
Clare Jones ◽  
Andrew Lawrence ◽  
Peter Wardman
Keyword(s):  
Cytochrome C ◽  
Hydroxyl Radical ◽  
Redox Regulation ◽  
Chemotherapeutic Agents ◽  
Tyrosyl Radical ◽  
Compound I ◽  
Definitive Evidence ◽  
Enhanced Production ◽  
Physico Chemical

The release of cytochrome c from mitochondria during apoptosis results in the enhanced production of superoxide radicals, which are converted to H2O2 by Mn-superoxide dismutase. We have been concerned with the role of cytochrome c/H2O2 in the induction of oxidative stress during apoptosis. Our initial studies showed that cytochrome c is a potent catalyst of 2′,7′-dichlorofluorescin oxidation, thereby explaining the increased rate of production of the fluorophore 2′,7′-dichlorofluorescein in apoptotic cells. Although it has been speculated that the oxidizing species may be a ferryl-haem intermediate, no definitive evidence for the formation of such a species has been reported. Alternatively, it is possible that the hydroxyl radical may be generated, as seen in the reaction of certain iron chelates with H2O2. By examining the effects of radical scavengers on 2′,7′-dichlorofluorescin oxidation by cytochrome c/H2O2, together with complementary EPR studies, we have demonstrated that the hydroxyl radical is not generated. Our findings point, instead, to the formation of a peroxidase compound I species, with one oxidizing equivalent present as an oxo-ferryl haem intermediate and the other as the tyrosyl radical identified by Barr and colleagues [Barr, Gunther, Deterding, Tomer and Mason (1996) J. Biol. Chem. 271, 15498-15503]. Studies with spin traps indicated that the oxo-ferryl haem is the active oxidant. These findings provide a physico-chemical basis for the redox changes that occur during apoptosis. Excessive changes (possibly catalysed by cytochrome c) may have implications for the redox regulation of cell death, including the sensitivity of tumour cells to chemotherapeutic agents.

PENGARUH PIJAT BAYI TERHADAP KUALITAS TIDUR BAYI UMUR 0 – 3 BULAN DI WILAYAH KERJA PUSKESMAS SOSIAL PALEMBANG TAHUN 2016

2019 ◽  
Vol 6 (2) ◽  
Author(s):  
Juliana Widyastuti Wahyuningsih Juliana Widyastuti Wahyuningsih
Keyword(s):  
Experimental Design ◽  
Growth And Development ◽  
Sampling Technique ◽  
P Value ◽  
Development Phase ◽  
Quality Of Sleep ◽  
Infant Massage ◽  
Before And After ◽  
The Brain

ABSTRAK Tidur merupakan kebutuhan yang harus terpenuhi terutama pada fase perkembangan karena selama tidur akan terjadi perkembangan otak maupun tubuh, sehingga gangguan tidur merupakan masalah yang akan menimbulkan dampak buruk terhadap pertumbuhan dan perkembangan bayi. Kualitas tidur bayi yang baik dapat diciptakan dengan memberikan pemijatan bayi secara rutin. Penelitian ini bertujuan untuk membuktikan bahwa pemijatan dapat mempengaruhi kualitas tidur bayi umur 0-3 bulan. Penelitian ini menggunakan desain penelitian Quasy Eksperimental dengan metode One Group Pretest-Postest. Sampel 22 bayi yang dipilih dengan tehnik Total Sampling yang di observasi sebelum dan sesudah diberikan pemijatan. Variabel yang diukur dalam penelitian ini adalah kualitas tidur bayi 0-3 bulan. Hasil penelitian menunjukkan bahwa ada pengaruh pijat bayi terhadap kualitas tidur bayi umur 0-3 bulan (p value  0,008 < α = 0,05).Berdasarkan hasil penelitian ini disarankan agar keluarga dan masyarakat memberikan pemijatan secara rutin dan mandiri untuk meningkatkan kebutuhan tidur bayi yang berkualitas.   ABSTRACT Sleep is a human necessity that must be met, especially in the development phase because during sleep will occur the brain and body developments, so that sleep disturbance is a problem that would cause adverse effects on infants’ growth and development. The good quality of sleep can be created by providing the infants massage routinely. This study aimed to prove that the massage could affect the quality of sleep on the 0-3 months old baby. This study used Quasy-experimental design with One Group Pretest-Posttest. The sample 22 infants selected by total sampling technique observed on before and after the massage. The variables measured in this study are the quality of sleep. The results of study indicate that there is an effect of infant massage to the sleep quality on 0-3 months old babies (p value 0,008 < α = 0,05).Based on the results of this study it recommended for the families and communities to provide infant massage regularly and independently to increase the quality of sleep on the baby.  

scholarly journals Systematical Engineering of Synthetic Yeast for Enhanced Production of Lycopene

2021 ◽  
Vol 8 (1) ◽  
pp. 14
Author(s):  
Yu Zhang ◽  
Tsan-Yu Chiu ◽  
Jin-Tao Zhang ◽  
Shu-Jie Wang ◽  
Shu-Wen Wang ◽  
...  
Keyword(s):  
Copy Number ◽  
Chromosome Rearrangement ◽  
Fold Increase ◽  
Host Strain ◽  
Target Product ◽  
Biosynthesis Pathway ◽  
Suitable Host ◽  
Enhanced Production ◽  
Condition Optimization ◽  
Heterologous Pathway

Synthetic biology allows the re-engineering of biological systems and promotes the development of bioengineering to a whole new level, showing great potential in biomanufacturing. Here, in order to make the heterologous lycopene biosynthesis pathway compatible with the host strain YSy 200, we evolved YSy200 using a unique Synthetic Chromosome Rearrangement and Modification by LoxP-mediated Evolution (SCRaMbLE) system that is built in the Sc2.0 synthetic yeast. By inducing SCRaMbLE, we successfully identified a host strain YSy201 that can be served as a suitable host to maintain the heterologous lycopene biosynthesis pathway. Then, we optimized the lycopene biosynthesis pathway and further integrated into the rDNA arrays of YSy201 to increase its copy number. In combination with culturing condition optimization, we successfully screened out the final yeast strain YSy222, which showed a 129.5-fold increase of lycopene yield in comparison with its parental strain. Our work shows that, the strategy of combining the engineering efforts on both the lycopene biosynthesis pathway and the host strain can improve the compatibility between the heterologous pathway and the host strain, which can further effectively increase the yield of the target product.

scholarly journals Advances in the Diagnosis and Treatment of Pediatric Acute Lymphoblastic Leukemia

2021 ◽  
Vol 10 (9) ◽  
pp. 1926
Author(s):  
Hiroto Inaba ◽  
Ching-Hon Pui
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Detailed Analysis ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Survival Rates ◽  
Treatment Strategies ◽  
Chemotherapeutic Agents ◽  
Response To Treatment ◽  
Pediatric Acute Lymphoblastic Leukemia ◽  
Therapeutic Implications

The outcomes of pediatric acute lymphoblastic leukemia (ALL) have improved remarkably during the last five decades. Such improvements were made possible by the incorporation of new diagnostic technologies, the effective administration of conventional chemotherapeutic agents, and the provision of better supportive care. With the 5-year survival rates now exceeding 90% in high-income countries, the goal for the next decade is to improve survival further toward 100% and to minimize treatment-related adverse effects. Based on genome-wide analyses, especially RNA-sequencing analyses, ALL can be classified into more than 20 B-lineage subtypes and more than 10 T-lineage subtypes with prognostic and therapeutic implications. Response to treatment is another critical prognostic factor, and detailed analysis of minimal residual disease can detect levels as low as one ALL cell among 1 million total cells. Such detailed analysis can facilitate the rational use of molecular targeted therapy and immunotherapy, which have emerged as new treatment strategies that can replace or reduce the use of conventional chemotherapy.

scholarly journals IL15RA and SMAD3 Genetic Variants Predict Overall Survival in Metastatic Colorectal Cancer Patients Treated with FOLFIRI Therapy: A New Paradigm

Cancers ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1705
Author(s):  
Elena De Mattia ◽  
Jerry Polesel ◽  
Rossana Roncato ◽  
Adrien Labriet ◽  
Alessia Bignucolo ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Overall Survival ◽  
Metastatic Colorectal Cancer ◽  
Cancer Patients ◽  
Prognostic Score ◽  
Chemotherapeutic Agents ◽  
Rank Test ◽  
P Value ◽  
Genetic Determinants ◽  
New Paradigm

A new paradigm in cancer chemotherapy derives from the interaction between chemotherapeutics, including irinotecan and 5-fluorouracil (5-FU), and the immune system. The patient’s immune response can modulate chemotherapy effectiveness, and, on the other hand, chemotherapeutic agents can foster tumor cell immunogenicity. On these grounds, the analysis of the cancer patients’ immunogenetic characteristics and their effect on survival after chemotherapy represent a new frontier. This study aims to identify genetic determinants in the immuno-related pathways predictive of overall survival (OS) after FOLFIRI (irinotecan, 5-FU, leucovorin) therapy. Two independent cohorts comprising a total of 335 patients with metastatic colorectal cancer (mCRC) homogeneously treated with first-line FOLFIRI were included in the study. The prognostic effect of 192 tagging genetic polymorphisms in 34 immune-related genes was evaluated using the bead array technology. The IL15RA rs7910212-C allele was associated with worse OS in both discovery (HR: 1.57, p = 0.0327, Bootstrap p-value = 0.0280) and replication (HR:1.71, p = 0.0411) cohorts. Conversely, SMAD3 rs7179840-C allele was associated with better OS in both discovery (HR:0.65, p = 0.0202, Bootstrap p-value = 0.0203) and replication (HR:0.61, p = 0.0216) cohorts. A genetic prognostic score was generated integrating IL15RA-rs7910212 and SMAD3-rs7179840 markers with inflammation-related prognostic polymorphisms we previously identified in the same study population (i.e., PXR [NR1I2]-rs1054190, VDR-rs7299460). The calculated genetic score successfully discriminated patients with different survival probabilities (p < 0.0001 log-rank test). These findings provide new insight on the prognostic value of genetic determinants, such as IL15RA and SMAD3 markers, and could offer a new decision tool to improve the clinical management of patients with mCRC receiving FOLFIRI.

scholarly journals Mating of 2 Laboratory Saccharomyces cerevisiae Strains Resulted in Enhanced Production of 2-Phenylethanol by Biotransformation of L-Phenylalanine

2017 ◽  
Vol 27 (2) ◽  
pp. 81-90 ◽  
Author(s):  
Jolanta Mierzejewska ◽  
Aleksandra Tymoszewska ◽  
Karolina Chreptowicz ◽  
Kamil Krol
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Batch Culture ◽  
Fold Increase ◽  
Biotechnological Production ◽  
Aromatic Alcohol ◽  
Enhanced Production ◽  
Yeast Strains ◽  
First Time

2-Phenylethanol (2-PE) is an aromatic alcohol with a rosy scent which is widely used in the food, fragrance, and cosmetic industries. Promising sources of natural 2-PE are microorganisms, especially yeasts, which can produce 2-PE by biosynthesis and biotransformation. Thus, the first challenging goal in the development of biotechnological production of 2-PE is searching for highly productive yeast strains. In the present work, 5 laboratory <i>Saccharomyces cerevisiae</i> strains were tested for the production of 2-PE. Thereafter, 2 of them were hybridized by a mating procedure and, as a result, a new diploid, <i>S. cerevisiae</i> AM1-d, was selected. Within the 72-h batch culture in a medium containing 5 g/L of <smlcap>L</smlcap>-phenylalanine, AM1-d produced 3.83 g/L of 2-PE in a shaking flask. In this way, we managed to select the diploid <i>S. cerevisiae</i> AM1-d strain, showing a 3- and 5-fold increase in 2-PE production in comparison to parental strains. Remarkably, the enhanced production of 2-PE by the hybrid of 2 yeast laboratory strains is demonstrated here for the first time.

Abstract 346: A Novel Interaction Between E2F and β-adrenergic Pathways Regulates Survival and Growth in Murine Myocardium

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Jennifer L Major ◽  
Maysoon Salih ◽  
Balwant S Tuana
Keyword(s):  
Protein Kinase ◽  
Left Ventricle ◽  
Fold Increase ◽  
Negative Regulator ◽  
P Value ◽  
Specific Expression ◽  
Cardiac Growth ◽  
Reduced Ejection Fraction ◽  
Hypertrophic Growth ◽  
Rb Pathway

The E2F/Pocket protein (Rb) pathway regulates cell growth, differentiation, and death by modulating gene expression. We previously examined this pathway in myocardium via manipulation of E2F6, which represses gene activity independently of Rb. Mice with cardiac specific expression of E2F6 develop dilated cardiomyopathy (DCM) without any signs of hypertrophic growth. We assessed the mechanisms of the apparent failure of compensatory growth as well as their response to the β-adrenergic agonist isoproterenol (iso). E2F6 transgenic (Tg) mice present with left ventricle dilation and significantly reduced ejection fraction as early as 2 weeks which persists into adulthood, but with no apparent increase in left ventricle weight: body weight (LVW:BW). E2F6-Tg mice treated with iso show double the increase in LVW: BW than their Wt counterparts (32% vs 16%, p-value: 0.007). Western blot revealed a specific activation of the β2-adrenergic pathway in Tg myocardium under basal conditions including a ~2-fold increase in β2-adrenergic receptors (β2-AR) (p-value: 8.9E-05), protein kinase A catalytic subunit (PKA-C) (p-value: 0.0176), activated c-SRC tyrosine-protein kinase (p-value: 0.0002), and an induction of the anti-apoptotic gene Bcl2. In contrast, a ~70% decrease in the cardiac growth regulator: AKT1 (p-value 0.0001) and a 4-fold increase in cyclic AMP dependent phosphodiesterase 4D (PDE4D), the negative regulator of PKA activity, was evident in Tg myocardium. The expression of E2F3 was de-regulated by E2F6, but was restored by iso while Rb expression was down-regulated. Thus deregulation of E2F/Rb pathway by E2F6 altered the β-adrenergic signaling pathway such that survival signaling was activated while hypertrophy was repressed resulting in the development of DCM without any increase in muscle mass. These data reveal a novel interplay between E2F and the β adrenergic pathway which regulate cardiac growth and fate.

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