Sequential and simultaneous statistical optimization by dynamic design of experiment for peptide overexpression in recombinant Escherichia coli View Full Text


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Article Info

DATE

2006-10

AUTHORS

Kwang-Min Lee, Chang-Hoon Rhee, Choong-Kyung Kang, Jung-Hoe Kim

ABSTRACT

The production of recombinant anti-HIV peptide, T-20, in Escherichia coli was optimized by statistical experimental designs (successive designs with multifactors) such as 2(4-1) fractional factorial, 2(3) full factorial, and 2(2) rotational central composite design in order. The effects of media compositions (glucose, NPK sources, MgSO4, and trace elements), induction level, induction timing (optical density at induction process), and induction duration (culture time after induction) on T-20 production were studied by using a statistical response surface method. A series of iterative experimental designs was employed to determine optimal fermentation conditions (media and process factors). Optimal ranges characterized by %T-20 (proportion of peptide to the total cell protein) were observed, narrowed down, and further investigated to determine the optimal combination of culture conditions, which was as follows: 9, 6, 10, and 1 mL of glucose, NPK sources, MgSO4, and trace elements, respectively, in a total of 100 mL of medium inducted at an OD of 0.55-0.75 with 0.7 mM isopropyl-beta-D-thiogalactopyranoside in an induction duration of 4 h. Under these conditions, up to 14% of T-20 was obtained. This statistical optimization allowed the production of T-20 to be increased more than twofold (from 6 to 14%) within a shorter induction duration (from 6 to 4 h) at the shake-flask scale. More... »

PAGES

59-80

References to SciGraph publications

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  • 2000-01. An optimization study of carotenoid production by Rhodotorula glutinis DBVPG 3853 from substrates containing concentrated rectified grape must as the sole carbohydrate source in JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
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  • 2006-05. Modeling and optimization of biopolymer (Polyhydroxyalkanoates) production from ice cream residue by novel statistical experimental design in APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
  • 2002-02. Expression of a thermostable restriction endonuclease in recombinant Escherichia coli cells and optimization of fermentation conditions in WORLD JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
  • Identifiers

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    http://scigraph.springernature.com/pub.10.1385/abab:135:1:59

    DOI

    http://dx.doi.org/10.1385/abab:135:1:59

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/17057256


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