Yibo Zhu1, Fagen Hu, Yingyue Zhu, Limei Wang, Bin Qi. 1. School of Biotechnology and Food Engineering, Changshu Institute of Technology, Changshu, 215500, Jiangsu, People's Republic of China, centuryrain@126.com.
Abstract
OBJECTIVES: The Tyr52 residue of D-lactate dehydrogenase (D-LDH) from Lactobacillus pentosus was replaced with small hydrophobic residues and overexpressed in E. coli BL21 (DE3) to enhance 3-phenyllactic acid (PLA) synthesis by whole-cell catalyst. RESULTS: Escherichia coli pET-28a-d-ldh produced 12.2 g PLA l(-1) in 3 h, with a molar conversion rate of 61 %, while E. coli pET-28a-d-ldh (Y52V) produced 15.6 g PLA l(-1), with a molar conversion rate of 77 %. This study demonstrates the feasibility of using engineered E. coli for PLA production from phenylpyruvate (PPA) and showed that site-directed mutagenesis of d-ldh markedly improved PLA yield and substrate conversion rate. CONCLUSION: This biocatalytic system is a promising platform for PLA production from PPA.
OBJECTIVES: The Tyr52 residue of D-lactate dehydrogenase (D-LDH) from Lactobacillus pentosus was replaced with small hydrophobic residues and overexpressed in E. coli BL21 (DE3) to enhance 3-phenyllactic acid (PLA) synthesis by whole-cell catalyst. RESULTS:Escherichia coli pET-28a-d-ldh produced 12.2 g PLA l(-1) in 3 h, with a molar conversion rate of 61 %, while E. coli pET-28a-d-ldh (Y52V) produced 15.6 g PLA l(-1), with a molar conversion rate of 77 %. This study demonstrates the feasibility of using engineered E. coli for PLA production from phenylpyruvate (PPA) and showed that site-directed mutagenesis of d-ldh markedly improved PLA yield and substrate conversion rate. CONCLUSION: This biocatalytic system is a promising platform for PLA production from PPA.
Authors: Sophie Klöppel; René Richarz; Daniel A Wirtz; Natalia Vasenda; Gabriele M König; Max Crüsemann Journal: Chembiochem Date: 2021-12-09 Impact factor: 3.461