Fed-Batch Production of Saccharomyces cerevisiae L-Asparaginase II by Recombinant Pichia pastoris MUTs Strain

Frontiers in Bioengineering and Biotechnology
David RodriguesAdalberto Jr Pessoa

Abstract

L-Asparaginase (ASNase) is used in the treatment of acute lymphoblastic leukemia, being produced and commercialized only from bacterial sources. Alternative Saccharomyces cerevisiae ASNase II coded by the ASP3 gene was biosynthesized by recombinant Pichia pastoris MUT s under the control of the AOX1 promoter, using different cultivation strategies. In particular, we applied multistage fed-batch cultivation divided in four distinct phases to produce ASNase II and determine the fermentation parameters, namely specific growth rate, biomass yield, and enzyme activity. Cultivation of recombinant P. pastoris under favorable conditions in a modified defined medium ensured a dry biomass concentration of 31 gdcw.L-1 during glycerol batch phase, corresponding to a biomass yield of 0.77 gdcw.g glycerol - 1 and a specific growth rate of 0.21 h-1. After 12 h of glycerol feeding under limiting conditions, cell concentration achieved 65 gdcw.L-1 while ethanol concentration was very low. During the phase of methanol induction, biomass concentration achieved 91 gdcw.L-1, periplasmic specific enzyme activity 37.1 U.g dcw - 1 , volumetric enzyme activity 3,315 U.L-1, overall enzyme volumetric productivity 31 U.L-1.h-1, while th...Continue Reading

References

Jun 1, 1975·Journal of Bacteriology·P C Dunlop, R J Roon
Jan 1, 1990·Annals of the New York Academy of Sciences·R A BrierleyR S Siegel
Jul 29, 1998·Methods in Molecular Biology·J StrattonM Meagher
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Sep 27, 2002·Current Opinion in Biotechnology·Geoff P Lin CereghinoJames M Cregg
Apr 4, 2003·Biotechnology and Bioengineering·M E BushellJ N Wardell
Feb 1, 1951·Journal of General Microbiology·W S WISE
Apr 1, 2004·Applied Biochemistry and Biotechnology·Maria Antonieta FerraraNei Pereira
Apr 17, 2004·Journal of Pediatric Hematology/oncology·Eduard H PanosyanUNKNOWN Children's Cancer Group Study CCG-1961
Jan 31, 2006·Applied Microbiology and Biotechnology·Wenhui ZhangMichael M Meagher
Oct 3, 2006·Critical Reviews in Oncology/hematology·Umesh K NartaWamik Azmi
Feb 6, 2009·Biotechnology Progress·Carl-Fredrik Mandenius, Anders Brundin
Oct 29, 2011·Microbial Cell Factories·Christian DietzschChristoph Herwig
Oct 17, 2013·Food Chemistry·Duniesky MartínezAlberto Del Monte-Martínez
Apr 20, 2014·Applied Microbiology and Biotechnology·Mudassar AhmadHelmut Schwab
Jun 2, 2015·Biotechnology Advances·V LooserK Kovar
Dec 24, 2015·Critical Reviews in Biotechnology·André Moreni LopesAdalberto Pessoa
Dec 30, 2015·Protein Expression and Purification·Luciana Facchinetti de Castro GirãoJonas Perales
Apr 27, 2016·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Chengcheng LiuMary V Relling
Dec 12, 2018·Journal of Dairy Science·Sabrina S SaboRicardo P S Oliveira

❮ Previous
Next ❯

Citations

Feb 14, 2020·Archives of Microbiology·Faizan MuneerHabibullah Nadeem
Jun 2, 2021·Applied Microbiology and Biotechnology·Daniel CastroValéria C Santos-Ebinuma

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
Protein Assay
glycosylation

Software Mentioned

Statistica

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.