Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

Shan Jiang; Mei Liu; Baojie Wang; Keyong Jiang; Lei Wang

doi:10.1007/s00343-011-0282-5

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Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

Updated：2023-05-16

- Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli
- Journal of Oceanology and Limnology Vol. 29, Issue 6, Pages: 1249-1259(2011)
- Affiliations：
  
  1. R&D Center of Marine Biotechnology Institute of Oceanology Chinese Academy of Sciences
  2. Graduate University of Chinese Academy of Sciences
- Author bio：
- Funds：
- DOI：10.1007/s00343-011-0282-5
  CLC：
- Online First：25 October 2011，
  
  Published：2011
- Accepted：
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Jiang, S., Liu, M., Wang, B. et al. Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli, ., Chin. J. Ocean. Limnol. 29, 1249–1259 (2011). https://doi.org/10.1007/s00343-011-0282-5
DOI：

Jiang, S., Liu, M., Wang, B. et al. Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli, ., Chin. J. Ocean. Limnol. 29, 1249–1259 (2011). https://doi.org/10.1007/s00343-011-0282-5 DOI：

摘要

Abstract

Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species. In a previous study

we isolated ALF genes from Chinese mitten crab

Eriocheir sinensis. In this study

we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Escherichia coli maintained in shake-flasks. In particular

we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design

and two significant screened factors

(NH4)2SO4 and KH2PO4

were further optimized via the central composite design (CCD). Based on the CCD analysis

we investigated the induction start-up time

the isopropylthio-D-galactoside (IPTG) concentration

the post-induction time

and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L (NH4)2SO4 and 3.18 g/L KH2PO4

with a cell optical density of 0.8 at 600 nm before induction

an IPTG concentration of 0.5 mmol/L

a post-induction temperature of 32.7°C

and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation

and determined that the soluble target protein constituted 10.5% of the total protein. Our identification of the economic medium composition

optimal culture conditions

and details of the fermentation process should facilitate the potential application of ALF for further research.

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