Heterologous expression of tyrosinase (MelC2) from Streptomyces avermitilis MA4680 in E
coli and its application for ortho -hydroxylation of resveratrol to produce piceatannol
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| 024 | 7 | 0 | |a 10.1007/s00253-015-6691-1 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6691-1 | ||
| 245 | 0 | 0 | |a Heterologous expression of tyrosinase (MelC2) from Streptomyces avermitilis MA4680 in E |h [Elektronische Daten] |b coli and its application for ortho -hydroxylation of resveratrol to produce piceatannol |c [Nahum Lee, Sang-Hyuk Lee, Kiheon Baek, Byung-Gee Kim] |
| 520 | 3 | |a Recombinant tyrosinase from Streptomyces avermitilis MA4680, MelC2 (gi:499291317), was heterologously expressed in Escherichia coli BL21 (DE3). The expression level of active MelC2 was increased by the codon-optimized MelC1 caddie protein (KP198295.1). By performing saturation mutagenesis of the Y91 residue of MelC1, it was found that aromatic residues such as Y, F, and W at the 91st position help produce a correctly folded conformation of MelC2. The recombinant MelC2 was utilized as a biocatalyst to convert trans-resveratrol into piceatannol. In order to improve the product yield through suppression of the formation of melanin, a by-product, an increase in the ratio of monooxygenation (k 1) to dioxygenation (k 2) of MelC2 is desirable. This was achieved by a combination of protein engineering and regeneration of NADH with glucose dehydrogenase (GDH). Saturation mutagenesis was performed at 15 residues within 8-Å radius from copper ions of MelC2. A total of 2760 mutants were examined (99.7% probability for NNK codon) and I41Y, a mutant, was screened. The ratio of k 1 to k 2 of the mutant increased sevenfold on tyrosine and fivefold on resveratrol, when compared to wild-type MelC2. As a result, the overall product yield from 500μM resveratrol in 50-mL reaction was 15.4% (77.4μM piceatannol), 1.7 times higher than wild type. When I41Y was incorporated with the NADH regeneration system, the total product yield was 58.0%, an eightfold increase (290.2μM of piceatannol). | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Tyrosinase |2 nationallicence | |
| 690 | 7 | |a Streptomyces avermitilis |2 nationallicence | |
| 690 | 7 | |a Resveratrol |2 nationallicence | |
| 690 | 7 | |a Piceatannol |2 nationallicence | |
| 700 | 1 | |a Lee |D Nahum |u School of Chemical and Biological Engineering, Seoul National University, 151-744, Seoul, South Korea |4 aut | |
| 700 | 1 | |a Lee |D Sang-Hyuk |u Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, 151-744, Seoul, South Korea |4 aut | |
| 700 | 1 | |a Baek |D Kiheon |u School of Chemical and Biological Engineering, Seoul National University, 151-744, Seoul, South Korea |4 aut | |
| 700 | 1 | |a Kim |D Byung-Gee |u School of Chemical and Biological Engineering, Seoul National University, 151-744, Seoul, South Korea |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/19(2015-10-01), 7915-7924 |x 0175-7598 |q 99:19<7915 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6691-1 |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/s00253-015-6691-1 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Lee |D Nahum |u School of Chemical and Biological Engineering, Seoul National University, 151-744, Seoul, South Korea |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Lee |D Sang-Hyuk |u Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, 151-744, Seoul, South Korea |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Baek |D Kiheon |u School of Chemical and Biological Engineering, Seoul National University, 151-744, Seoul, South Korea |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Kim |D Byung-Gee |u School of Chemical and Biological Engineering, Seoul National University, 151-744, Seoul, South Korea |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/19(2015-10-01), 7915-7924 |x 0175-7598 |q 99:19<7915 |1 2015 |2 99 |o 253 | ||