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Effects of pulse feeding of beet molasses on recombinant benzaldehyde lyase production by Escherichia coli BL21(DE3)

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Abstract

The effect of fed-batch operation (FBO) strategy was investigated using pretreated-beet molasses, containing galactose that induces the lac promoter, on benzaldehyde lyase (BAL) production by recombinant Escherichia coli BL21(DE3)pLySs. After batch cultivation with 30 g l−1 pretreated-beet molasses consisting of 7.5 g l−1 glucose and 7.5 g l−1 fructose, three FBO strategies were applied at dissolved oxygen (=40%) cascade to air-flow rate. In FBO1 when air-flow rate decreased considerably, feed was given to the system in pulses in such a way that pretreated-beet molasses concentration increased by 10 kg m−3 (containing 2.5 g l−1 glucose+2.5 g l−1 fructose); however, decrease in air-flow rate demonstrated only the absence of glucose but not fructose. Thus, in FBO2 when fructose and glucose were completely utilized, pretreated-beet molasses was pulse-fed and its concentration increased by 10 g l−1. In FBO3 with the decreased amount of pretreated-beet molasses (6 g l−1), shift response time from glucose to fructose consumption was avoided, and glucose and fructose consumptions were well correlated with air-flow rate, and the highest C X (8.04 g l−1) and BAL (2,315 U ml−1) production were obtained (t = 24 h) with the highest substrate yield on cell and product formation.

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Abbreviations

A :

Benzaldehyde lyase volumetric activity, U ml−1

A S :

Benzaldehyde lyase specific activity, U mg−1 DW

C OA :

Organic acid concentration, g l−1

C AA :

Acetic acid concentration, g l−1

C DO :

Dissolved oxygen concentration, mmol l−1

\( C_{\text{DO}}^{*} \) :

Oxygen saturation concentration, mmol l−1

C G :

Glucose concentration, g l−1

C F :

Fructose concentration, g l−1

C X :

Cell concentration, g l−1

K L a :

Overall liquid phase mass transfer coefficient, s−1

N :

Agitation rate, min–1

m 0 :

Maintenance for oxygen consumption, g oxygen g−1 dry cell weight h−1

m S :

Maintenance coefficients for substrate, g substrate g−1 dry cell weight h−1

Q :

Volumetric air feed rate, l min−1

q O :

Specific oxygen uptake rate, g g –1 DW h−1

q S :

Specific substrate consumption rate, g g –1 DW h−1

r 0 :

Oxygen uptake rate, mmol l−1 s−1

T :

Bioreaction medium temperature, °C

t :

Bioreactor cultivation time, h

U:

One unit of an enzyme

V :

Volume of the bioreactor, l

V R :

Volume of the bioreaction medium, l

Y X/S :

Yield of cell on substrate, g g−1

Y X/O :

Yield of cell on oxygen, g g−1

Y S/O :

Yield of substrate on oxygen, g g−1

µ :

Specific cell growth rate, h−1

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Acknowledgments

This work was supported by the Scientific and Technical Research Council of Turkey (TUBITAK: MAG-104 M258) and METU Research Fund.

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Authors and Affiliations

  1. Department of Chemical Engineering, Industrial Biotechnology and Metabolic Engineering Laboratory, Middle East Technical University, 06531, Ankara, Turkey

    Pınar Çalık & Hande Levent

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  1. Pınar Çalık
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  2. Hande Levent
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Correspondence to Pınar Çalık.

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Çalık, P., Levent, H. Effects of pulse feeding of beet molasses on recombinant benzaldehyde lyase production by Escherichia coli BL21(DE3). Appl Microbiol Biotechnol 85, 65–73 (2009). https://doi.org/10.1007/s00253-009-2060-2

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  • DOI: https://doi.org/10.1007/s00253-009-2060-2

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