Feeding strategy design for recombinant human growth hormone production by Bacillus subtilis

Date

2015-10-01

Author

Sahin, Burcu
Ozturk, Sibel
Çalık, Pınar
Ozdamar, Tuncer H.

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

194
views

0
downloads

Defined and semi-defined medium-based feeding strategies were developed to enhance recombinant human growth hormone (rhGH) production by Bacillus subtilis BGSC-1A178 (scoC (-)) strain carrying pMK4::pre(subC)::hGH. Defined medium-based feeding strategies were designed by exponential feeding of glucose and (NH4)(2)HPO4 at two pre-determined specific growth rates, A mu (0) = 0.10 and 0.17 h(-1). Semi-defined medium-based feeding strategies were designed by exponential feeding of substrate solution consisting of glucose, (NH4)(2)HPO4, peptone, and trace salt solution (PTM1) at three pre-determined specific growth rates, A mu (0) = 0.10, 0.17, and 0.25 h(-1). At all the strategies applied, transition cultivation time from batch to fed-batch operation was t (T) = 4 h. The highest rhGH concentration was obtained as C (rhGH) = 0.5 g L-1 with semi-defined medium-based feeding strategy designed with A mu (0) = 0.25 h(-1) using feed substrate stock solution containing 200 g L-1 glucose, 117 g L-1 (NH4)(2)HPO4, 100 g L-1 peptone, and 5 mL L-1 PTM1 at t = 22 h when the cell concentration reached to C (X) = 8.29 g L-1. The overall product and cell yields on glucose were obtained as = 7.21 mg g(-1) and = 0.12 g g(-1), respectively. The results indicate the requirement of designing continuous feed stream in fed-batch production to enhance rhGH production by r-B. subtilis.

Subject Keywords

Fed-Batch Operation, Exponential Feeding, Glucose, Semi-Defined Medium, Human Growth Hormone, Bacillus Subtilis

URI

https://hdl.handle.net/11511/42571

Journal

BIOPROCESS AND BIOSYSTEMS ENGINEERING

DOI

https://doi.org/10.1007/s00449-015-1426-3

Collections

Department of Chemical Engineering, Article

Suggestions

OpenMETU
Core

Feeding strategy development for human growth hormone production by Pichia pastoris Bozkurt, Bahar; Çalık, Pınar; Özdamar, Tunçer H.; Department of Biotechnology (2012) In this study, recombinant human growth hormone (rhGH) production by Pichia pastoris-Mut+ strain was improved by designing feeding strategies which were applied in the production phase of the bioreactor operations. During the bio-reactor experiments the cell growth, sorbitol and methanol consumptions, recom-binant hGH production, alcohol oxidase (AOX) activity, the by-products protease and organic acid concentrations were followed and analyzed. In this context, in the first part of the study, three bioreact...
Glucose-based feeding strategy development in recombinant human growth hormone production by ichia pastoris for semi-batch bioreactor operation Keskin, Abdullah; Çalık, Pınar; Özdamar, Tunçer; Department of Chemical Engineering (2014) The aim of this study was to develop an effective feeding strategy to enhance recombinant human growth hormone (rhGH) production by Pichia pastoris Mut+ strain harboring pGAPZαA::hGH which is integrated to glyceraldehyde-3-phosphate dehydrogenase gene (GAP) locus of P. pastoris. In this context, eight semi-batch bioreactor experiments were conducted at pilot-scale; their batch phase was same in which glycerol was used as the sole carbon source. For semi-batch operation phase, eight feeding strategies were d...
Expression System for Recombinant Human Growth Hormone Production from Bacillus subtilis ÖZDAMAR, HASAN TUNÇER; Sentuerk, Birguel; Yilmaz, Oezge Deniz; Calik, Guezide; Celik, Eda; Çalık, Pınar (2009-01-01) We demonstrate for the first time, an expression system mimicking serine alkaline protease synthesis and secretion, producing native form of human growth hormone (hGH) from Bacillus subtilis. A hybrid-gene of two DNA fragments, i.e., signal (pre-) DNA sequence of B. licheniformis serine alkaline protease gene (subC) and cDNA encoding hGH, were cloned into pMK4 and expressed under deg-promoter in B. subtilis. Recombinant-hGH (rhGH) produced by B. subtilis carrying pMK4::pre(subC)::hGH was secreted. N-termina...
Human growth hormone production by metabolically engineered b. subtilis : feeding strategy development Gökbudak, Burcu; Çalık, Pınar; Özdamar, Tunçer H.; Department of Chemical Engineering (2013) In this study, the aim is to develop an effective feeding strategy for therapeutically important protein, recombinant human growth hormone (rhGH) production by Bacillus subtilis scoC knockout (scoC-) strain carrying pMK4::pre(subC)::hGH plasmid. In the first part of the research program, laboratory scale air filtered shake bioreactor experiments were conducted to optimize the production conditions. Thereafter, semi-batch production processes at pilot scale bioreactor system were performed by designing expon...
Semi-defined medium based feeding strategy development for human growth hormone production by recombinant bacillus subtilis/ Öztürk, Sibel; Çalık, Pınar; Özdamar, Tunçer H.; Department of Biotechnology (2014) In this study, the aim was to develop an efficient semi-defined medium based continuous feeding strategy for recombinant human growth hormone (rhGH) production by Bacillus subtilis 1A178 scoC knockout (scoC-) strain carrying pMK4::pre(subC)::hGH plasmid. In this context, firstly laboratory-scale air-filtered shake bioreactor experiments were carried out in order to investigate the effects of peptone as the complex carbon and nitrogen source on the cell growth and rhGH production both in the absence and the ...

Citation Formats

B. Sahin, S. Ozturk, P. Çalık, and T. H. Ozdamar, “Feeding strategy design for recombinant human growth hormone production by Bacillus subtilis,” BIOPROCESS AND BIOSYSTEMS ENGINEERING, pp. 1855–1865, 2015, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/42571.