Purification and Biochemical Characterization of a Tyrosine Phenol-lyase from Morganella morganii
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Purification and Biochemical Characterization of a Tyrosine Phenol-lyase from Morganella morganii Hang-Qin Zhu 1,2 & Xiao-Ling Tang 1,2 & Ren-Chao Zheng 1,2 & Yu-Guo Zheng 1,2 Received: 5 November 2019 / Accepted: 12 March 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
Tyrosine phenol-lyase (TPL) is a valuable and cost-effective biocatalyst for the biosynthesis of L-tyrosine and its derivatives, which are valuable intermediates in the pharmaceutical industry. A TPL from Morganella morganii (Mm-TPL) was overexpressed in Escherichia coli and characterized. Mm-TPL was determined as a homotetramer with molecular weight of 52 kDa per subunit. Its optimal temperature and pH for βelimination of L-tyrosine were 45 °C and pH 8.5, respectively. Mm-TPL manifested strict substrate specificity for the reverse reaction of β-elimination and ortho- and metasubstituted phenols with small steric size were preferred substrates. The enzyme showed excellent catalytic performance for synthesis of L-tyrosine, 3-fluoro-L-tyrosine, and LDOPA with a yield of 98.1%, 95.1%, and 87.2%, respectively. Furthermore, the fed-batch bioprocess displayed space-time yields of 9.6 g L−1 h−1 for L-tyrosine and 4.2 g L−1 h−1 for 3-fluoro-L-tyrosine with a yield of 67.4 g L−1 and 29.5 g L−1, respectively. These results demonstrated the great potential of Mm-TPL for industrial application. Keywords Tyrosine phenol-lyase . The reverse reaction of β-elimination . Morganella morganii . L-tyrosine and its derivatives . Phenolic compounds
* Ren-Chao Zheng [email protected] Hang-Qin Zhu [email protected] Xiao-Ling Tang [email protected] Yu-Guo Zheng [email protected]
1
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People’s Republic of China
2
Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, People’s Republic of China
Applied Biochemistry and Biotechnology
Introduction Tyrosine phenol-lyase (TPL, EC 4.1.99.2), a pyridoxal 5′-phosphate (PLP)-dependent enzyme, catalyzes the reversible β-elimination of L-tyrosine to produce phenol, pyruvate, and ammonium (Scheme 1) [1]. Apart from the physiological reaction, TPL catalyzes a variety of reactions including the reversible β-elimination and β-substitution reactions of other αamino acids [2, 3]. Some of these TPL-mediated reactions have attracted much attention due to the potential applications in synthesis of L-tyrosine and its ring-substituted analogues. L-tyrosine and its derivatives are important compounds in the pharmaceutical and nutrient fields. L-tyrosine is a valuable compound used as a dietary supplement, and the precursor for several neurotransmitters including dopamine, epinephrine, and norepinephrine, which were important drugs for neurological and cancer disease [4, 5]. 3,4-Dihydroxyphenyl-L-alanine (LDOPA) is the first line for Parkinson’s disease treatmen
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