Enhanced Interferon-α2b Production in Periplasmic Space of Escherichia coli through Medium Optimization using Response Surface Method

The influence of different carbon and nitrogen sources on growth of recombinant Escherichia coli and human interferon-α2b (IFN-α2b) production in periplasmic space was studied in shake flask culture. A statistical method based on Plackett-Burman design was used to screen the main medium components that greatly influenced the performance of the fermentation process. The optimization of medium was performed using response surface methodology (RSM) where three critical factors (glucose, yeast extract and peptone) were optimized using central composite design. The highest yield of periplasmic recombinant human interferon-α2b (PrIFN-α2b) (335.8 μg/L) was predicted to be obtained in optimized medium containing 5.47 g/L glucose, 55.24 g/L yeast extract and 42.27 g/L peptone.. The production of IFN-α2b in periplasmic space in optimized medium was about 2.5, 11.7 and 124.4 times higher than Terrific broth (TB), Luria-Bertani (LB), and minimal medium (M9), respectively.

Download Full-text

OVAT Analysis and Response Surface Methodology Based on Nutrient Sources for Optimization of Pigment Production in The Marine-Derived Fungus Talaromyces albobiverticillius 30548 Submerged Fermentation

Marine Drugs ◽

10.3390/md19050248 ◽

2021 ◽

Vol 19 (5) ◽

pp. 248

Author(s):

Mekala Venkatachalam ◽

Alain Shum-Chéong-Sing ◽

Yanis Caro ◽

Laurent Dufossé ◽

Mireille Fouillaud

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Yeast Extract ◽

Maximum Yield ◽

Nitrogen Sources ◽

Industrial Applications ◽

Fungal Species ◽

Pigment Production ◽

Interactive Effects ◽

Medium Components

Pigment production from filamentous fungi is gaining interest due to the diversity of fungal species, the variety of compounds synthesized, and the possibility of controlled massive productions. The Talaromyces species produce a large panel of metabolites, including Monascus-like azaphilone pigments, with potential use as natural colorants in industrial applications. Optimizing pigment production from fungal strains grown on different carbon and nitrogen sources, using statistical methods, is widespread nowadays. The present work is the first in an attempt to optimize pigments production in a culture of the marine-derived T. albobiverticillius 30548, under the influence of several nutrients sources. Nutrient combinations were screened through the one-variable-at-a-time (OVAT) analysis. Sucrose combined with yeast extract provided a maximum yield of orange pigments (OPY) and red pigments (RPY) (respectively, 1.39 g/L quinizarin equivalent and 2.44 g/L Red Yeast pigment equivalent), as well as higher dry biomass (DBW) (6.60 g/L). Significant medium components (yeast extract, K2HPO4 and MgSO4·7H2O) were also identified from one-variable-at-a-time (OVAT) analysis for pigment and biomass production. A five-level central composite design (CCD) and a response surface methodology (RSM) were applied to evaluate the optimal concentrations and interactive effects between selected nutrients. The experimental results were well fitted with the chosen statistical model. The predicted maximum response for OPY (1.43 g/L), RPY (2.59 g/L), and DBW (15.98 g/L) were obtained at 3 g/L yeast extract, 1 g/L K2HPO4, and 0.2 g/L MgSO4·7H2O. Such optimization is of great significance for the selection of key nutrients and their concentrations in order to increase the pigment production at a pilot or industrial scale.

Download Full-text

The role of lac operon and lac repressor in the induction using lactose for the expression of periplasmic human interferon-α2b by Escherichia coli

Annals of Microbiology ◽

10.1007/s13213-011-0394-3 ◽

2011 ◽

Vol 62 (4) ◽

pp. 1427-1435 ◽

Cited By ~ 2

Author(s):

Joo Shun Tan ◽

Ramakrishnan Nagasundara Ramanan ◽

Tau Chuan Ling ◽

Shuhaimi Mustafa ◽

Arbakariya B. Ariff

Keyword(s):

Escherichia Coli ◽

Lac Repressor ◽

Human Interferon ◽

Lac Operon ◽

Interferon Α2b

Download Full-text

Optimization of osmotic shock process variables for enhancement of the release of periplasmic interferon-α2b from Escherichia coli using response surface method

Process Biochemistry ◽

10.1016/j.procbio.2009.09.007 ◽

2010 ◽

Vol 45 (2) ◽

pp. 196-202 ◽

Cited By ~ 19

Author(s):

Ramakrishnan Nagasundara Ramanan ◽

Joo Shun Tan ◽

Mohd Shamzi Mohamed ◽

Tau Chuan Ling ◽

Beng Ti Tey ◽

...

Keyword(s):

Escherichia Coli ◽

Response Surface ◽

Response Surface Method ◽

Osmotic Shock ◽

Process Variables ◽

Shock Process ◽

Surface Method ◽

Interferon Α2b

Download Full-text

Purification and conformational properties of a human interferon α2b produced in Escherichia coli

Biotechnology and Applied Biochemistry ◽

10.1042/ba20010001 ◽

2001 ◽

Vol 33 (3) ◽

pp. 173 ◽

Cited By ~ 25

Author(s):

Alejandro Beldarraín ◽

Yai Cruz ◽

Oscar Cruz ◽

Monica Navarro ◽

Miriela Gil

Keyword(s):

Escherichia Coli ◽

Human Interferon ◽

Conformational Properties ◽

Interferon Α2b

Download Full-text

Optimization of Medium Components for β-Glucosidase Production from Aspergillus Niger by Response Surface Methodology

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.419.328 ◽

2013 ◽

Vol 419 ◽

pp. 328-333 ◽

Cited By ~ 1

Author(s):

Chao Zhang ◽

Rui Huang ◽

Hui Tian ◽

Ru Ming Zhao ◽

Fa Shun Yu ◽

...

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Yeast Extract ◽

Influence Factors ◽

Mutual Effect ◽

Tween 80 ◽

Fermentation Medium ◽

Medium Composition ◽

Malt Extract ◽

Medium Components

β-Glucosidase is the key enzyme for the utilization of lignocellulose.But the commercial β-glucosidase can’t be produced. This paper focuses on the study of the β-glucosidase fermentation process.The fermentation medium components for β-glucosidase production from Aspergil lusniger was optimized by response surface methodology (RSM). Firstly, the three of the most important influence factors yeast extract, MnSO4•H2O and MgSO4•7H2O was obtained from Plackett-Burman design screening. Then the path of steepest ascent experiment was adopted to approach the optimal region of the medium composition. Lastly, the optimal concentration and mutual effect of three factors were predicted by RSM. The results showed that the best medium composition was Malt extract 18g/L, Yeast extract 3.22g/L, KH2PO4 3g/L, MnSO4•H2O 0.58mM, Tween-80 0.5mL/L and MgSO4•7H2O 0.23g/L. Under these fermentation conditions, the activity of β-glucosidase was up to 7.33IU/mL with increasing 23.2% than before.

Download Full-text

Optimization of human interferon gamma production in Escherichia coli by response surface methodology

Biotechnology and Bioprocess Engineering ◽

10.1007/s12257-007-0126-5 ◽

2008 ◽

Vol 13 (1) ◽

pp. 7-13 ◽

Cited By ~ 11

Author(s):

Victor E. Balderas Hernández ◽

Luz M. T. Paz Maldonado ◽

Emilio Medina Rivero ◽

Ana P. Barba de la Rosa ◽

Leandro G. Ordoñez Acevedo ◽

...

Keyword(s):

Escherichia Coli ◽

Response Surface Methodology ◽

Response Surface ◽

Interferon Gamma ◽

Human Interferon

Download Full-text

Optimization of L-asparaginase production from Escherichia coli using response surface methodology

Vietnam Journal of Biotechnology ◽

10.15625/1811-4989/16/4/10861 ◽

2020 ◽

Vol 16 (4) ◽

pp. 767-775

Author(s):

Nguyen Thi Hien Trang ◽

Le Thanh Hoang ◽

Do Thi Tuyen

Keyword(s):

Escherichia Coli ◽

Response Surface Methodology ◽

Recombinant Protein ◽

Response Surface ◽

Lymphoblastic Leukemia ◽

Quadratic Model ◽

Shake Flask Culture ◽

Cultivation Conditions ◽

Maldi Tof ◽

Optimized Conditions

Among the antitumor drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. In previous study, the L-asparaginase from Erwinia chrysanthermy was expressed in Escherichia coli BL21(DE3). The recombinant L-asparaginase was produced from recombinant E.coli BL21(DE3) under different cultivation conditions (inducer concentration, inoculum concentration and KH2PO4 concentration). The optimized conditions by response surface methodology using face centered central composite design. The analysis of variance coupled with larger value of R2 (0.9) showed that the quadratic model used for the prediction was highly significant (p < 0.05). Under the optimized conditions, the model produced L-asparaginase activity of 123.74 U/ml at 1.03 mM IPTG, 3% (v/v) inoculum and 0.5% (w/v) KH2PO4. Recombinant protein was purified by two step using gel filtration and DEAE chromatography. The purified L-asparaginase had a molecular mass of 37 kDa with specific activity of 462 U/mg and identified by MALDI-TOF mass spectrometry. Results of MALDI-TOF analysis confirmed that recombinant protein was L-asparaginase II. Recombinant L-asparaginase has antiproliferative activity with K562 cell line. In conclusion, this study has innovatively developed cultivation conditions for better production of recombinant L-asparaginase in shake flask culture.

Download Full-text

Optimization of Fermentation Conditions for Recombinant Human Interferon Beta Production by Escherichia coli Using the Response Surface Methodology

Jundishapur Journal of Microbiology ◽

10.5812/jjm.8(4)2015.16236 ◽

2015 ◽

Vol 8 (4) ◽

Cited By ~ 18

Author(s):

Mohammad Hossein Morowvat ◽

Valiollah Babaeipour ◽

Hamid Rajabi Memari ◽

Hossein Vahidi

Keyword(s):

Escherichia Coli ◽

Response Surface Methodology ◽

Response Surface ◽

Interferon Beta ◽

Human Interferon ◽

Fermentation Conditions ◽

Optimization Of Fermentation ◽

Recombinant Human Interferon Beta

Download Full-text

Optimization of the Sporogenous Medium of Irpex Lacteus Using Response Surface Methodology

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.343-344.832 ◽

2011 ◽

Vol 343-344 ◽

pp. 832-837

Author(s):

Na Zhang ◽

Qing Yan Chen ◽

Ruo Chuan Pan ◽

Xue Feng Yao ◽

Yan Feng Wang ◽

...

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Yeast Extract ◽

Fermentation Broth ◽

Sporulation Medium ◽

Irpex Lacteus ◽

Normal Medium ◽

Medium Components ◽

Five Factors ◽

Central Composite

Response surface methodology (RSM) was used to optimize sporulation medium components for improving spores concentration in I. lacteus fermentation broth. Five factors of sporulation medium components, glucose (X1), peptone (X2), yeast extract (X3) MgSO4·7H2O (X4) and VB1 (X5), were chosen to optimize through a five-level, five-factor central composite design (CCD) base on single factor experiments. The concentration of medium components which influenced the sporulation of I. lacteus was further optimized using RSM. The results show that The medium that consists of 4.15 g/L peptone, 2.04 g/L yeast extract, 5.10 g/L glucose, 0.17 g/L MgSO4·7H2O and 0.03 g/L VB1 enhance spores concentration significantly . The final spores concentration (2.19×105 /mL) by using the optimized medium was about five times higher than normal medium.

Download Full-text