Yield Optimization of A Heterologously Expressed Novel Mouse Macrophage Protein

Parveen Pahuja^1, Alagiri Srinivasan² and Munish Puri^1,3*

1Fermentation and Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala, India. 2Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India. 3Centre for Chemistry and Biotechnology, Deakin University, Geelong Technology Precinct, Victoria - 3217, Australia.

DOI : http://dx.doi.org/10.13005/bbra/1108

ABSTRACT: The large scale whole-genome sequencing projects have resulted in large numbers of un-characterized and un-annotated protein sequences. This presents an opportunity and a challenge to characterise these novel protein sequences. Structural biology has become a widely accepted methodology to help assign functions to such proteins, complementing other cellular and biochemical studies. However, most of these studies require the target protein to be produced in large quantities and in a highly pure and soluble state. The present study is an attempt to maximise production of a recombinant mouse macrophage protein (rMMP) over-expressed heterologously in Escherichia coli. Highest production of biomass and total protein (6.6 mg mL-1) was observed at 37 oC. Maximum cell disruption (89%) was observed during freeze-thawing and subsequent ultrasonication.

KEYWORDS: E.coli; cell disruption; Ultra-sonication; Lysozyme; Freeze-thawing; Pretreatment

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