Hydrolysis of polyethyleneterephthalate by p-nitrobenzylesterase from Bacillus subtilis
Ribitsch, D; Heumann, S; Trotscha, E; Herrero Acero, E; Greimel, K; Leber, R; Birner-Gruenberger, R; Deller, S; Eiteljoerg, I; Remler, P; Weber, T; Siegert, P; Maurer, KH; Donelli, I; Freddi, G; Schwab, H; Guebitz, GM
| HERO ID | 1455197 |
|---|---|
| In Press | No |
| Year | 2011 |
| Title | Hydrolysis of polyethyleneterephthalate by p-nitrobenzylesterase from Bacillus subtilis |
| Authors | Ribitsch, D; Heumann, S; Trotscha, E; Herrero Acero, E; Greimel, K; Leber, R; Birner-Gruenberger, R; Deller, S; Eiteljoerg, I; Remler, P; Weber, T; Siegert, P; Maurer, KH; Donelli, I; Freddi, G; Schwab, H; Guebitz, GM |
| Journal | Biotechnology Progress |
| Volume | 27 |
| Issue | 4 |
| Page Numbers | 951-960 |
| Abstract | From a screening on agar plates with bis(benzoyloxyethyl) terephthalate (3PET), a Bacillus subtilis p-nitrobenzylesterase (BsEstB) was isolated and demonstrated to hydrolyze polyethyleneterephthalate (PET). PET-hydrolase active strains produced clearing zones and led to the release of the 3PET hydrolysis products terephthalic acid (TA), benzoic acid (BA), 2-hydroxyethyl benzoate (HEB), and mono-(2-hydroxyethyl) terephthalate (MHET) in 3PET supplemented liquid cultures. The 3PET-hydrolase was isolated from non-denaturating polyacrylamide gels using fluorescein diacetate (FDA) and identified as BsEstB by LC-MS/MS analysis. BsEstB was expressed in Escherichia coli with C-terminally fused StrepTag II for purification. The tagged enzyme had a molecular mass of 55.2 kDa and a specific activity of 77 U/mg on p-nitrophenyl acetate and 108 U/mg on p-nitrophenyl butyrate. BsEstB was most active at 40°C and pH 7.0 and stable for several days at pH 7.0 and 37°C while the half-life times decreased to 3 days at 40°C and only 6 h at 45°C. From 3PET, BsEstB released TA, MHET, and BA, but neither bis(2-hydroxyethyl) terephthalate (BHET) nor hydroxyethylbenzoate (HEB). The k(cat) values decreased with increasing complexity of the substrate from 6 and 8 (s-1) for p-nitrophenyl-acetate (4NPA) and p-nitrophenyl-butyrate (4NPB), respectively, to 0.14 (s-1) for bis(2-hydroxyethyl) terephthalate (BHET). The enzyme hydrolyzed PET films releasing TA and MHET with a concomitant decrease of the water-contact angle (WCA) from 68.2° ± 1.7° to 62.6° ± 1.1° due to formation of novel hydroxyl and carboxyl groups. These data correlated with a fluorescence emission intensity increase seen for the enzyme treated sample after derivatization with 2-(bromomethyl)naphthalene. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011. |
| Doi | 10.1002/btpr.610 |
| Pmid | 21574267 |
| Wosid | WOS:000294107600006 |
| Url | <Go to ISI>://WOS:000294107600006 |
| Is Certified Translation | No |
| Dupe Override | No |
| Is Public | Yes |
| Language Text | English |
| Keyword | Water Resources Abstracts; Biotechnology and Bioengineering Abstracts; Benzoic acid; Biotechnology; Bacillus subtilis; Escherichia coli; Radioactive Half-life; Fluorescence; fluorescein diacetate; pH effects; Bacillus; Enzymes; Purification; Terephthalic acid; Data processing; Hydrolysis; p-Nitrophenyl; Liquid culture; Hydrogen Ion Concentration; Acetic acid; W 30940:Products; SW 0810:General |