Active tyrosine phenol-lyase aggregates induced by terminally attached functional peptides in Escherichia coli
- PDF / 1,813,926 Bytes
- 9 Pages / 595.276 x 790.866 pts Page_size
- 40 Downloads / 149 Views
BIOCATALYSIS - ORIGINAL PAPER
Active tyrosine phenol‑lyase aggregates induced by terminally attached functional peptides in Escherichia coli Hongmei Han1,2,3 · Weizhu Zeng2,3,4 · Guoqiang Zhang1,2,3 · Jingwen Zhou1,2,3,4 Received: 7 March 2020 / Accepted: 20 July 2020 © The Author(s) 2020
Abstract The formation of inclusion bodies (IBs) without enzyme activity in bacterial research is generally undesirable. Researchers have attempted to recovery the enzyme activities of IBs, which are commonly known as active IBs. Tyrosine phenol-lyase (TPL) is an important enzyme that can convert pyruvate and phenol into 3,4-dihydroxyphenyl-l-alanine (L-DOPA) and IBs of TPL can commonly occur. To induce the correct folding and recover the enzyme activity of the IBs, peptides, such as ELK16, DKL6, L6KD, ELP10, ELP20, L6K2, EAK16, 18A, and GFIL16, were fused to the carboxyl terminus of TPL. The results showed that aggregate particles of TPL-DKL6, TPL-ELP10, TPL-EAK16, TPL-18A, and TPL-GFIL16 improved the enzyme activity by 40.9%, 50.7%, 48.9%, 86.6%, and 97.9%, respectively. The peptides TPL-DKL6, TPL-EAK16, TPL-18A, and TPL-GFIL16 displayed significantly improved thermostability compared with TPL. L-DOPA titer of TPL-ELP10, TPLEAK16, TPL-18A, and TPL-GFIL16, with cells reaching 37.8 g/L, 53.8 g/L, 37.5 g/L, and 29.1 g/L, had an improvement of 111%, 201%, 109%, and 63%, respectively. A higher activity and L-DOPA titer of the TPL-EAK16 could be valuable for its industrial application to biosynthesize L-DOPA. Keywords Self-assembling peptide · Tyrosine phenol-lyase · Active inclusion bodies · Thermostability · L-DOPA
Introduction Tyrosine phenol-lyase (TPL) (EC4.1.99.2), a tetrameric enzyme, can catalyze stereospecific isotope exchange of α-protons of various amino acids with Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10295-020-02294-4) contains supplementary material, which is available to authorized users. * Jingwen Zhou [email protected] 1
National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, Jiangsu, China
2
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, Jiangsu, China
3
The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi 214122, Jiangsu, China
4
Jiangsu Provisional Research Center for Bioactive Product Processing Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, Jiangsu, China
pyridoxal-5′-phosphate (PLP) as the cofactor [20]. TPL has been mainly isolated and characterized from bacteria, such as Citrobacter freundii [38,46], Erwinia herbicola [58], and Fusobacterium nucleatum [62]. Monovalent cations, K + or NH4+ are necessary for achieving high activity levels of TPL [33]. The biosynthesis based on the enzyme activity of TPL has attracted attention for its application in the production of enant
Data Loading...
