Reconstruction of lactate utilization system in Pseudomonas putida KT2440: a novel biocatalyst for l-2-hydroxy-carboxylate production.
As an important method for building blocks synthesis, whole cell biocatalysis is hindered by some shortcomings such as unpredictability of reactions, utilization of opportunistic pathogen, and side reactions. Due to its biological and extensively studied genetic background, Pseudomonas putida KT2440 is viewed as a promising host for construction of efficient biocatalysts. After analysis and reconstruction of the lactate utilization system in the P. putida strain, a novel biocatalyst that only exhibited NAD-independent D-lactate dehydrogenase activity was prepared and used in L-2-hydroxy-carboxylates production. Since the side reaction catalyzed by the NAD-independent L-lactate dehydrogenase was eliminated in whole cells of recombinant P. putida KT2440, two important L-2-hydroxy-carboxylates (L-lactate and L-2-hydroxybutyrate) were produced in high yield and high optical purity by kinetic resolution of racemic 2-hydroxy carboxylic acids. The results highlight the promise in biocatalysis by the biotechnologically important organism P. putida KT2440 through genomic analysis and recombination.
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Yujiao
Y
Wang
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Min
M
Lv
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Yingxin
Y
Zhang
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Xieyue
X
Xiao
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Tianyi
T
Jiang
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Wen
W
Zhang
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Chunhui
C
Hu
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Chao
C
Gao
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
Cuiqing
C
Ma
1] State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China [2] State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
Ping
P
Xu
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D055162:Biocatalysis; D015964:Gene Expression Regulation, Bacterial; D006885:Hydroxybutyrates / Q000378:metabolism*; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D045022:Lactate Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D007773:Lactates / Q000378:metabolism*; D060847:Metabolic Engineering; D009876:Operon; D016958:Pseudomonas putida / Q000235:genetics / Q000378:metabolism*; D013237:Stereoisomerism
D001426:Bacterial Proteins; D006885:Hydroxybutyrates; D007773:Lactates; D045022:Lactate Dehydrogenases; D007770:L-Lactate Dehydrogenase; C071195:D-lactate dehydrogenase; C031570:2-hydroxybutyric acid