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

Your Location：

Home >

Browse articles >

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

OriginalPaper | 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
- Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli
- 海洋湖沼学报（英文） 2011年29卷第6期页码：1249-1259
- 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：
  
  Supported by the National High Technology Research and Development Program of China (863 Program) (No. 2006AA100311)
- DOI：10.1007/s00343-011-0282-5
  中图分类号：
- 网络首发：2011-10-25，
  
  纸质出版：2011
- Accepted：
Scan QR Code
Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli[J]. 海洋湖沼学报（英文）, 2011,29(6):1249-1259.

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
Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli[J]. 海洋湖沼学报（英文）, 2011,29(6):1249-1259. DOI： 10.1007/s00343-011-0282-5.

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.

关键词

Keywords

references

Belloto R J, Dean A M, Moustafa M A, Molokhia A M, Gouda M W, Sokoloski T D. 1985. Statistical techniques applied to solubility predictions and pharmaceutical formulations: an approach to problem solving using mixture response surface methodology. Int. J. Pharm., 23: 195–207.

Beres D L, Hawkins D M. 2001. Plackett-Burman techniques for sensitivity analysis of many-parametered models. Ecol. Model., 141: 171–183.

Chen H M, Wang W, Smith D, Chan S C. 1997. Effects of the anti-bacterial peptide cecropin B and its analogs, cecropins B-1 and B-2, on liposomes, bacteria, and cancer cells. BBA — General Subjects, 1 336: 171–179.

Choi J H, Keum K C, Lee S Y. 2006. Production of recombinant proteins by high cell density culture of Escherichia coli. Chem. Eng. Sci., 61: 876–885.

Ge Y, MacDonald D, Henry M M, Hait H I, Nelson K A, Lipsky B A, Zasloff M A, Holroyd K J. 1999. In vitro susceptibility to pexiganan of bacteria isolated from infected diabetic foot ulcers. Diagn. Micr. Infec. Dis., 35: 45–53.

Gross P S, Bartlett T C, Browdy C L, Chapman R W, Warr G W. 2001. Immune gene discovery by expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific white shrimp, Litopenaeus vannamei, and the Atlantic white shrimp, L. setiferus. Dev. Comp. Immunol., 25: 565–577.

Kang C H, Wang J X, Zhao X F, Yang X M, Shao H L, Xiang J H. 2004. Molecular cloning and expression analysis of Ch-penaeidin, an antimicrobial peptide from Chinese shrimp, Fenneropenaeus chinensis. Fish Shellfish Immun., 16: 513–525.

Lachman E, Pitsoe S, Gaffin S. 1984. Anti-lipolysaccharide immnotherapy in management of septic shock of obstetric and gynaecological origin. Lancet, 323:981–983.

Li C H, Zhao J M, Song L S, Mu C K, Zhang H, Gai Y C, Qiu L M, Yu Y D, Ni D J, Xing K Z. 2008a. Molecular cloning, genomic organization and functional analysis of an anti-lipopolysaccharide factor from Chinese mitten crab Eriocheir sinensis. Dev. Comp. Immunol., 32: 784–794.

Li X, Xu T C, Ma X H, Guo K P, Kai L, Zhao Y H, Jia X M, Ma Y. 2008b. Optimization of culture conditions for production of cis-epoxysuccinic acid hydrolase using response surface methodology. Bioresource Technol., 99: 5 391–5 396.

Lo P K, Hassan O, Ahmad A, Mahadi N M, Illias R M. 2007. Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology. Enzyme Microb. Tech., 40: 1 256–1 263.

Mekata T, Sudhakaran R, Okugawa S, Kono T, Sakai M, Itami T. 2010. Molecular cloning and transcriptional analysis of a newly identified anti-lipopolysaccharide factor gene in kuruma shrimp, Marsupenaeus japonicus. Lett. Appl. Microbiol., 50: 112–119.

Miller G L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem., 31: 426–428.

Morita T, Ohtsubo S, Nakamura T, Tanaka S, Iwanaga S, Ohashi K, Niwa M. 1985. Isolation and biological activities of limulus anticoagulant (anti-LPS factor) which interacts with lipopolysaccharide (LPS). J. Biochem., 97: 1 611–1 620.

Murby M, Uhlen M, Stahl S. 1996. Upstream strategies to minimize proteolytic degradation upon recombinant production in Escherichia coli. Protein Expres. Purif., 7: 129–136.

Myers R, Montgomery D, Anderson-Cook C. 2009. Response surface methodology: process and product optimization using designed experiments. John Wiley & Sons Inc.

Nikerel I E, Toksoy E, Kirdar B, YildIrIm R. 2005. Optimizing medium composition for TaqI endonuclease production by recombinant Escherichia coli cells using response surface methodology. Process Biochem., 40: 1 633–1 639.

Riesenberg D. 1991. High-cell-density cultivation of Escherichia coli. Curr. Opin. Biotech., 2: 380–384.

Rubinchik E, Dugourd D, Algara T, Pasetka C, Friedland H D. 2009. Antimicrobial and antifungal activities of a novel cationic antimicrobial peptide, omiganan, in experimental skin colonisation models. Int. J. Antimicrob. Ag., 34: 457–461.

Sambrook J, Russel D W. 2001. Molecular Cloning: A Laboratory Manual. Spring Harbor Laboratory Press.

Sarmasik A, Chen T T. 2003. Bactericidal activity of cecropin B and cecropin P1 expressed in fish cells (CHSE-214): application in controlling fish bacterial pathogens. Aquaculture, 220: 183–194.

Schein C H. 1989. Production of soluble recombinant proteins in bacteria. Nat. Biotech., 7: 1 141–1 149.

Shiloach J, Fass R. 2005. Growing E. coli to high cell density—A historical perspective on method development. Biotechno. Adv., 23: 345–357.

Steiner H, Hultmark D, Engstrom A, Bennich H, Boman H G. 1981. Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature, 292: 246–248.

Supungul P, Klinbunga S, Pichyangkura R, Jitrapakdee S, Hirono I, Aoki T, Tassanakajon A. 2002. Identification of immune-related genes in hemocytes of black tiger shrimp (Penaeus monodon). Mar. Biotechnol., 4: 487–494.

Tanaka S, Nakamura T, Morita T, Iwanaga S. 1982. Anti-LPS factor: An anticoagulant which inhibits the endotoxinmediated activation of coagulation system. Biochem. Bioph. Res. Co., 105: 717–723.

Trotti A, Garden A, Warde P, Symonds P, Langer C, Redman R, Pajak T F, Fleming T R, Henke M, Bourhis J, Rosenthal D I, Junor E, Cmelak A, Sheehan F, Pulliam J, Devitt-Risse P, Fuchs H, Chambers M, O’sullivan B, Ang K K. 2004. A multinational, randomized phase iii trial of iseganan hcl oral solution for reducing the severity of oral mucositis in patients receiving radiotherapy for head-and-neck malignancy. Int. J. Radiat. Oncol., 58: 674–681.

Villaverde A, Mar Carrió M. 2003. Protein aggregation in recombinant bacteria: biological role of inclusion bodies. Biotechnol. Lett., 25: 1 385–1 395.

Wang D N, Chen L, Liu J W, He Z Y, Zhang W J, Wu X F. 2001. The native gene of anti-LPS factor from Tachypleus tridentatus: Cloning, expression and its bacteriostatic activity in vitro. Protein Peptide Lett., 8: 273–280.

Wang Y H, Jing C F, Yang B, Mainda G, Dong M L, Xu A L. 2005. Production of a new sea anemone neurotoxin by recombinant Escherichia coli: Optimization of culture conditions using response surface methodology. Process Biochem., 40: 2 721–2 728.

Wood T K, Peretti S W. 1991. Effect of chemically-induced, cloned-gene expression on protein synthesis in E. coli. Biotechnol. Bioeng., 38: 397–412.

浏览量

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

工具集

关联资源

Effects of nutrients and light intensity on the growth and biochemical composition of a marine microalga Odontella aurita

Optimization of fermentation conditions for 1,3-propanediol production by marine Klebsiella pneumonia HSL4 using response surface methodology

双转子轴间滑油颗粒运动轨迹参数化模型

高压涡轮封严流与主流相互作用的机理研究

一种涡轮基组合动力的整机低速风洞试验研究