Coexpression of Lactobacillus brevis ADH with GDH or G6PDH in Arxula adeninivorans for the synthesis of 1-(R)-phenylethanol
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| 024 | 7 | 0 | |a 10.1007/s00253-014-6297-z |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-014-6297-z | ||
| 245 | 0 | 0 | |a Coexpression of Lactobacillus brevis ADH with GDH or G6PDH in Arxula adeninivorans for the synthesis of 1-(R)-phenylethanol |h [Elektronische Daten] |c [Marion Rauter, Alexandra Prokoph, Jakub Kasprzak, Karin Becker, Keith Baronian, Rüdiger Bode, Gotthard Kunze, H.- Vorbrodt] |
| 520 | 3 | |a The yeast Arxula adeninivorans was used for the overexpression of an ADH gene of Lactobacillus brevis coding for (R)-specific alcohol dehydrogenase (LbADH) to synthesise enantiomerically pure 1-(R)-phenylethanol. Glucose dehydrogenase gene from Bacillus megaterium (BmGDH) or glucose 6-phosphate dehydrogenase of Bacillus pumilus (BpG6PDH) were coexpressed in Arxula to regenerate the cofactor NADPH by oxidising glucose or glucose 6-phosphate. The yeast strain expressing LbADH and BpG6PDH produced 5200U l-1 ADH and 370U l-1 G6PDH activity, whereas the strain expressing LbADH and BmGDH produced 2700U l-1 ADH and 170U l-1 GDH activity. However, the crude extract of both strains reduced 40mM acetophenone to pure 1-(R)-phenylethanol with an enantiomeric excess (ee) of >99% in 60min without detectable by-products. An increase in yield was achieved using immobilised crude extracts (IEs), Triton X-100 permeabilised cells (PCs) and permeabilised immobilised cells (PICs) with PICs being most stable with GDH regeneration over 52 cycles. Even though the activity and synthesis rate of 1-(R)-phenylethanol with the BpG6PDH and LbADH coexpressing strain was higher, the BmGDH-LbADH strain was more stable over successive reaction cycles. This, combined with its higher total turnover number (TTN) of 391 mol product per mole NADP+, makes it the preferred strain for continuous reaction systems. The initial non-optimised semi-continuous reaction produced 9.74gl−1day−1 or 406gkg−1 dry cell weight (dcw) day−1 isolated 1-(R)-phenylethanol with an ee of 100% and a TTN of 206mol product per mole NADP+. In conclusion, A. adeninivorans is a promising host for LbADH and BpG6PDH or BmGDH production and offers a simple method for the production of enantiomerically pure alcohols. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2014 | ||
| 690 | 7 | |a 1-(R)-Phenylethanol |2 nationallicence | |
| 690 | 7 | |a Lactobacillus brevis ADH |2 nationallicence | |
| 690 | 7 | |a Bacillus megaterium GDH |2 nationallicence | |
| 690 | 7 | |a Bacillus pumilus BpG6PDH |2 nationallicence | |
| 690 | 7 | |a Arxula adeninivorans |2 nationallicence | |
| 690 | 7 | |a Cell permeabilisation |2 nationallicence | |
| 690 | 7 | |a Calcium alginate immobilisation |2 nationallicence | |
| 700 | 1 | |a Rauter |D Marion |u Orgentis Chemicals GmbH, Bahnhofstr. 3-5, 06466, Gatersleben, Germany |4 aut | |
| 700 | 1 | |a Prokoph |D Alexandra |u Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466, Gatersleben, Saxony-Anhalt, Germany |4 aut | |
| 700 | 1 | |a Kasprzak |D Jakub |u Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466, Gatersleben, Saxony-Anhalt, Germany |4 aut | |
| 700 | 1 | |a Becker |D Karin |u Orgentis Chemicals GmbH, Bahnhofstr. 3-5, 06466, Gatersleben, Germany |4 aut | |
| 700 | 1 | |a Baronian |D Keith |u School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand |4 aut | |
| 700 | 1 | |a Bode |D Rüdiger |u Institute of Microbiology, University of Greifswald, Felix-Hausdorffstr. 4, 17487, Greifswald, Germany |4 aut | |
| 700 | 1 | |a Kunze |D Gotthard |u Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466, Gatersleben, Saxony-Anhalt, Germany |4 aut | |
| 700 | 1 | |a Vorbrodt |D H.-. |u Orgentis Chemicals GmbH, Bahnhofstr. 3-5, 06466, Gatersleben, Germany |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/11(2015-06-01), 4723-4733 |x 0175-7598 |q 99:11<4723 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-014-6297-z |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-014-6297-z |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Rauter |D Marion |u Orgentis Chemicals GmbH, Bahnhofstr. 3-5, 06466, Gatersleben, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Prokoph |D Alexandra |u Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466, Gatersleben, Saxony-Anhalt, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Kasprzak |D Jakub |u Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466, Gatersleben, Saxony-Anhalt, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Becker |D Karin |u Orgentis Chemicals GmbH, Bahnhofstr. 3-5, 06466, Gatersleben, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Baronian |D Keith |u School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Bode |D Rüdiger |u Institute of Microbiology, University of Greifswald, Felix-Hausdorffstr. 4, 17487, Greifswald, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Kunze |D Gotthard |u Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466, Gatersleben, Saxony-Anhalt, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Vorbrodt |D H.-. |u Orgentis Chemicals GmbH, Bahnhofstr. 3-5, 06466, Gatersleben, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/11(2015-06-01), 4723-4733 |x 0175-7598 |q 99:11<4723 |1 2015 |2 99 |o 253 | ||