Hyper-production And Stability Of Human Serum Albumin In Pichia Pastoris Through A Combination Of Medium Design And Genetic Strategies | 97613
ISSN: 2155-952X

Journal of Biotechnology & Biomaterials
Open Access

Like us on:

Our Group organises 3000+ Global Conferenceseries Events every year across USA, Europe & Asia with support from 1000 more scientific Societies and Publishes 700+ Open Access Journals which contains over 50000 eminent personalities, reputed scientists as editorial board members.

Open Access Journals gaining more Readers and Citations
700 Journals and 15,000,000 Readers Each Journal is getting 25,000+ Readers

This Readership is 10 times more when compared to other Subscription Journals (Source: Google Analytics)
Recommended Conferences

2nd International Conference on Biotechnology and Healthcare

Auckland, New Zealand

6th International Conference on Vaccines and Immunology

Geneva, Switzerland

3rd European Congress on Virology

Geneva, Switzerland
Google scholar citation report
Citations : 1600

Journal of Biotechnology & Biomaterials received 1600 citations as per google scholar report

Indexed In
  • Index Copernicus
  • Google Scholar
  • Sherpa Romeo
  • Open J Gate
  • Genamics JournalSeek
  • Academic Keys
  • ResearchBible
  • China National Knowledge Infrastructure (CNKI)
  • Access to Global Online Research in Agriculture (AGORA)
  • Electronic Journals Library
  • RefSeek
  • Hamdard University
  • OCLC- WorldCat
  • SWB online catalog
  • Virtual Library of Biology (vifabio)
  • Publons
  • Geneva Foundation for Medical Education and Research
  • Euro Pub
Share This Page

Hyper-production and stability of human serum albumin in Pichia pastoris through a combination of medium design and genetic strategies

21st European Biotechnology Congress

Saroj Mishra and Nitu Maity

Indian Institute of Technology Delhi, India

ScientificTracks Abstracts: J Biotechnol Biomater

DOI: 10.4172/2155-952X-C5-100

Human serum albumin (HSA) is an important therapeutic recommended for treatment against trauma, burn injury, hypoproteinemia, hypoalbumenia as well as for maintenance of homeostasis, transportation of hormones and microelements in blood. In this study, we report medium design and genetic strategies that lead to high production of this protein in the culture spernatant of Pichia pastoris. The codon-optimized gene for HSA was cloned downstream of ????factor secretory signal sequence and the mature HSA was secreted in the culture supenatant of P. pastoris under the control of alcohol oxidase 1 promoter. Extracellular protein level of 0.12, 0.40, 1.2 g/L were obtained in the un-optimized medium for 1-copy, 2-copy and 3-copy expression casettes respectively at shake flask level. Factors affecting production were identified which included initial peptone concentration, methanol concentration and temperature, amongst many other (pH, aeration, sorbitol concentration, initial inoculum) investigated parameters. A three level factorial design named central composite design using Plackett Burman response surface methodology was used to optimize the medium which lead to levels of protein up to 0.075, 0.40 and 0.98 g/L total extracellular protein for 1,2 and 3-copy constructs respectively. Under these conditions, HSA produced was stable and free of other contaminating proteins in the culture supernatant. A detailed transcriptome analysis of the recombinant P. pastoris, cultivated on unoptimized and optimized medium lead to identification of several protein coding transcripts which were up-regulated and helped in efficient HSA production and secretion. These were mapped to biochemical activities linked (and not restricted to) to carbon, nitrogen metabolism, gene transcription, protein transport and secretion. Additional genetic strategies applied included modification of signal sequences. Application of optimized medium to these mutants lead to stable production of HSA with reduced proteolytic degradation of the synthesized protein. This illustrated the robustness of the designed medium with a production of over 2 g/L protein at shakeflask level. An understanding of the underlying mechanisms is likely to play significant role in use of Pichia system for production of heterologous proteins.

Saroj Mishra completed her PhD from City University of NewYork, USA followed by Post-doctoral research at Institute Pasteur, Paris, France, VTT Biotechnical Laboratory, Espoo, Finland and University of California, Davis, USA. She has published more than 87 papers in reputed journals and leads a large group of scientists working in the area of enviromental biotechnology, whole cell biotransformation and recombinant therapeutics.

E-mail: [email protected]