caa a22 4500 605499470 CHVBK 20210128100551.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6757-0 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6757-0 Production of phenylpyruvic acid from l -phenylalanine using an l -amino acid deaminase from Proteus mirabilis [Elektronische Daten] comparison of enzymatic and whole-cell biotransformation approaches [Ying Hou, Gazi Hossain, Jianghua Li, Hyun-dong Shin, Long Liu, Guocheng Du] Phenylpyruvic acid (PPA) is an important organic acid that has a wide range of applications. In this study, the membrane-bound l-amino acid deaminase (l-AAD) gene from Proteus mirabilis KCTC 2566 was expressed in Escherichia coli BL21(DE3) and then the l-AAD was purified. After that, we used the purified enzyme and the recombinant E. coli whole-cell biocatalyst to produce PPA via a one-step biotransformation from l-phenylalanine. l-AAD was solubilized from the membrane and purified 52-fold with an overall yield of 13%, which corresponded to a specific activity of 0.94 ± 0.01μmol PPA min−1·mg−1. Then, the biotransformation conditions for the pure enzyme and the whole-cell biocatalyst were optimized. The maximal production was 2.6 ± 0.1g·L−1 (specific activity of 1.02 ± 0.02μmol PPA min−1·mg−1 protein, 86.7 ± 5% mass conversion rate, and 1.04g·L−1·h−1 productivity) and 3.3 ± 0.2gL−1 (specific activity of 0.013 ± 0.003μmol PPA min−1·mg−1 protein, 82.5 ± 4% mass conversion rate, and 0.55g·L−1·h−1 productivity) for the pure enzyme and whole-cell biocatalyst, respectively. Comparative studies of the enzymatic and whole-cell biotransformation were performed in terms of specific activity, production, conversion, productivity, stability, need of external cofactors, and recycling. We have developed two eco-friendly and efficient approaches for PPA production. The strategy described herein may aid the biotransformational synthesis of other α-keto acids from their corresponding amino acids. Springer-Verlag Berlin Heidelberg, 2015 l -Amino acid deaminase nationallicence Phenylpyruvic acid nationallicence Proteus mirabilis nationallicence Whole-cell biotransformation nationallicence Hou Ying Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut Hossain Gazi Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut Li Jianghua Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut Shin Hyun-dong School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 30332, Atlanta, USA aut Liu Long Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut Du Guocheng Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8391-8402 0175-7598 99:20<8391 2015 99 253 https://doi.org/10.1007/s00253-015-6757-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6757-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hou Ying Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut NATIONALLICENCE 700 1- Hossain Gazi Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut NATIONALLICENCE 700 1- Li Jianghua Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut NATIONALLICENCE 700 1- Shin Hyun-dong School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 30332, Atlanta, USA aut NATIONALLICENCE 700 1- Liu Long Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut NATIONALLICENCE 700 1- Du Guocheng Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8391-8402 0175-7598 99:20<8391 2015 99 253