Production of excreted human epidermal growth factor (hEGF) by an efficient recombinant Escherichia coli system

Sivakesava, S.; Xu, ZN; Chen, Y-H.; Hackett, Jim; Huang, RC; Lam, E.; Lam, TL; Siu, KL; Wong, Roderick; Wong, Wan Keung

doi:10.1016/s0032-9592(99)00013-8

Cited by 52 publications

(37 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In shake flask culture incubated on rotary shaker, the agitation speed of 200 rpm was found optimal for growth of recombinant E. coli. [21,22] The xylanase production obtained in this study (2122.5 U mL −1 ) was significantly higher than those reported in the literature. For example, the highest xylanase production obtained in submerged cultivation of Aspergillus niger B03 was 996.3 U mL −1 [23] and 592.7 U g/substrate by Trichoderma longibrachiatum [24] .…”

Section: Production Of Intracellular Xylanase By E Colicontrasting

confidence: 64%

Kinetics of Xylanase Fermentation by Recombinant <i>Escherichia coli</i> DH5α in Shake Flask Culture

Rusli¹

2009

American Journal of Biochemistry and Biotechnology

View full text Add to dashboard Cite

Problem statement: Interest in xylanase enzyme application has markedly increased in pulp and paper processing industries. The switch to xylanase-producing recombinant Escherichia coli DH5α pTP510 is seen here as an economic alternative towards higher productivity and easier downstream purification. Modeling of E. coli DH5α growth and enzyme secretion is thus desired for future optimization in fermentation process. Approach: Kinetics of intracellular xylanase fermentation by a recombinant E. coli DH5α was studied in shake flask culture. The effect of different medium formulations (complex, minimal and defined), initial pH (6.5, 7.0, 7.4 and 8.0) and agitation speeds (150, 200 and 250 rpm) on cell growth and xylanase production were evaluated. Mathematical models based on Logistic and Luedeking-Piret equations had been proposed to describe the microbial growth and xylanase production. Results: Highest xylanase production was obtained in defined medium. Based on medium formulation, the highest cell concentration (4.59 g L −1 ) and xylanase production (2, 122.5 U mL −1 ) was obtained when (NH 4 ) 2 HPO 4 was used as the main nitrogen source, with an adjustment of the initial pH to 7.4 and agitation speed of 200 rpm. The maximum specific growth rate (µ max ), growth associated xylanase production coefficient (α) and non-growth associated xylanase production coefficient (β) was 0.41 h −1 , 474.26 U mg cell −1 and 0 U mg cell −1 h −1 , respectively. Conclusion: Xylanase production was growth associated process and the enzyme secretion was greatly dependent on cell concentration and the specific growth rate of E. coli DH5α.

show abstract

Section: Production Of Intracellular Xylanase By E Colicontrasting

confidence: 64%

Kinetics of Xylanase Fermentation by Recombinant <i>Escherichia coli</i> DH5α in Shake Flask Culture

Rusli¹

2009

American Journal of Biochemistry and Biotechnology

View full text Add to dashboard Cite

show abstract

“…A major drawback of the fusion approach is the formation of rEGF as variants possessing different peptide lengths (Table 1) [11][12][13][14][15][16][17][18][19][20][21][22]. Moreover, the EGF derivatives were commonly shown to exhibit lower levels of bioactivity and stability [17,[22][23][24].…”

Section: Approaches Of Expressing Recombinant Egfmentioning

confidence: 99%

“…Moreover, the EGF derivatives were commonly shown to exhibit lower levels of bioactivity and stability [17,[22][23][24]. Later on, pragmatic approaches were developed for the production of not only bioactive and stable rEGF, but also a product available at a much reduced price [11][12].…”

Section: Approaches Of Expressing Recombinant Egfmentioning

confidence: 99%

Review Article: Reasons for Underrating the Potential of Human Epidermal Growth Factor in Medical Applications

Wong¹,

Kl²,

Cc³

et al. 2017

JAPLR

View full text Add to dashboard Cite

Human epidermal growth factor (EGF) is a functionally versatile polypeptide comprising 53 amino acids (aa). Due to the ability of EGF to stimulate epidermal growth and proliferation, numerous research studies have been conducted to explore its potential applications in cosmetic, skin care and medication. The emergence of recombinant DNA technology has facilitated the production of a great variety of recombinant EGF (rEGF) isoforms. However, increasing evidence supports that rEGF isoforms exhibit different levels of stability and potency. This review discusses how the less bioactive rEGF isoforms may mystify the identity and even the functional properties of authentic rEGF, which shares the same primary structure as native EGF. The identity crisis, the time-consuming, costly and labor-intensive patenting and drug regulatory processes act together to result in underrating the functional applications of authentic rEGF, which have been shown to be effective and safe in promoting treatments of a variety of skin disorders such as diabetic foot ulcers (DFU).

show abstract

“…The basic composition of MMBL (modified MBL) medium was (g/L): glucose, (variable); tryptone, 10.0; yeast extract, 10.0; NaCl, 10.0; glycerol, 2.0; K 2 HPO 4 , 0.5; KH 2 PO 4 , 2.5; (NH 4 ) 2 HPO 4 , 2.0; MgSO 4 , 1.0; trace metal solution, 1.0 ml/L; ampicillin, 70 mg/L (when required) (SivaKesava et al, 1999 BO 3 , 0.5; HCl (37%, w/v), 37 ml/L. Plackett-Burman experiments were carried out to find the key factors which influence the plasmid productivity.…”

Section: Culture Mediamentioning

confidence: 99%

“…In the past twenty years, many efficient strategies for recombinant-cell fermentation had been developed to attain high productivity of proteins in E. coli, B. subtillis, yeast, even in animal cell culture systems (SivaKesava et al, 1999;Ebisu et al, 1992;Cheng et al, 1997;Churgay et al, 1997). However, few papers were focused on the plasmid DNA production in recombinant E. coli fermentation process (Lahijani et al, 1996;O'Kennedy et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy

Shen

Chen

et al. 2005

J Zheijang Univ Sci B

View full text Add to dashboard Cite

Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system.

show abstract

Production of excreted human epidermal growth factor (hEGF) by an efficient recombinant Escherichia coli system

Cited by 52 publications

References 27 publications

Kinetics of Xylanase Fermentation by Recombinant <i>Escherichia coli</i> DH5α in Shake Flask Culture

Kinetics of Xylanase Fermentation by Recombinant <i>Escherichia coli</i> DH5α in Shake Flask Culture

Review Article: Reasons for Underrating the Potential of Human Epidermal Growth Factor in Medical Applications

Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy

Contact Info

Product

Resources

About