One-Pot Biosynthesis of 3-Aminopropionic Acid from Fumaric Acid Using Recombinant Bacillus megaterium Containing a Linear Dual-Enzyme Cascade

Author(s):  
Subbi Rami Reddy Tadi ◽  
Ganesh Nehru ◽  
Senthilkumar Sivaprakasam
2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Yuanyuan Jiang ◽  
Zhong Li ◽  
Shanmin Zheng ◽  
Huifang Xu ◽  
Yongjin J. Zhou ◽  
...  

2011 ◽  
Vol 13 (10) ◽  
pp. 2895 ◽  
Author(s):  
Lara Babich ◽  
Lieke J. C. van Hemert ◽  
Aleksandra Bury ◽  
Aloysius F. Hartog ◽  
Pierpaolo Falcicchio ◽  
...  

2017 ◽  
Vol 115 (1) ◽  
pp. 192-205 ◽  
Author(s):  
Thomas F.T. Rexer ◽  
Anna Schildbach ◽  
Jan Klapproth ◽  
Angelika Schierhorn ◽  
Reza Mahour ◽  
...  

2020 ◽  
Vol 68 (34) ◽  
pp. 9188-9194
Author(s):  
Zonglin Li ◽  
Xiao Ning ◽  
Yiran Zhao ◽  
Xiaodan Zhang ◽  
Chun Xiao ◽  
...  

Catalysts ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1470
Author(s):  
Fei Liu ◽  
Junping Zhou ◽  
Meijuan Xu ◽  
Taowei Yang ◽  
Minglong Shao ◽  
...  

Unusual α-amino acids (UAAs) are important fundamental building blocks and play a key role in medicinal chemistry. Here, we constructed a hydrogen-borrowing dual-enzyme cascade for efficient synthesis of UAAs from α-hydroxy acids (α-HAs). D-mandelate dehydrogenase from Lactobacillus brevis (LbMDH) was screened for the catalysis of α-HAs to α-keto acids but with low activity towards aliphatic α-HAs. Therefore, we rational engineered LbMDH to improve its activity towards aliphatic α-HAs. The substitution of residue Leu243 located in the substrate entrance channel with nonpolar amino acids like Met, Trp, and Ile significantly influenced the enzyme activity towards different α-HAs. Compared with wild type (WT), variant L243W showed 103 U/mg activity towards D-α-hydroxybutyric acid, 1.7 times of the WT’s 60.2 U/mg, while its activity towards D-mandelic acid decreased. Variant L243M showed 2.3 times activity towards D-mandelic acid compared to WT, and its half-life at 40 °C increased to 150.2 h comparing with 98.5 h of WT. By combining LbMDH with L-leucine dehydrogenase from Bacillus cereus, the synthesis of structurally diverse range of UAAs from α-HAs was constructed. We achieved 90.7% conversion for L-phenylglycine production and 66.7% conversion for L-α-aminobutyric acid production. This redox self-sufficient cascade provided high catalytic efficiency and generated pure products.


Processes ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 932
Author(s):  
Zhongkui Li ◽  
Xiangsong Chen ◽  
Zhijian Ni ◽  
Lixia Yuan ◽  
Lijie Sun ◽  
...  

Sialyllactose (SL) is one of the most important acidic oligosaccharides in human milk, which plays an important role in the health of infants. In this work, an efficient multi-enzyme cascade was developed in a single whole cell to produce 3′-SL. We constructed two compatible plasmids with double cloning sites to co-express four genes. Different combinations were assessed to verify the optimal catalytic ability. Then, the conversion temperature, pH, and stability under the optimal temperature and pH were investigated. Moreover, the optimal conversion conditions and surfactant concentration were determined. By using the optimal conditions (35 °C, pH 7.0, 20 mM polyphosphate, 10 mM cytidine monophosphate (CMP), 20 mM MgCl2), 25 mL and 4 L conversion systems were carried out to produce 3′-SL. Similar results were obtained between different volume conversion reactions, which led the maximum production of 3′-SL to reach 53 mM from 54.2 mM of sialic acid (SA) in the 25 mL system and 52.8 mM of 3′-SL from 53.8 mM of SA in the 4 L system. These encouraging results demonstrate that the developed single whole-cell multi-enzyme system exhibits great potential and economic competitiveness for the manufacture of 3′-SL.


Membranes ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 28
Author(s):  
Fatin Nasreen Ahmad Rizal Lim ◽  
Fauziah Marpani ◽  
Victoria Eliz Anak Dilol ◽  
Syazana Mohamad Pauzi ◽  
Nur Hidayati Othman ◽  
...  

Multi-enzyme cascade catalysis involved three types of dehydrogenase enzymes, namely, formate dehydrogenase (FDH), formaldehyde dehydrogenase (FaldDH), alcohol dehydrogenase (ADH), and an equimolar electron donor, nicotinamide adenine dinucleotide (NADH), assisting the reaction is an interesting pathway to reduce thermodynamically stable molecules of CO2 from the atmosphere. The biocatalytic sequence is interesting because it operates under mild reaction conditions (low temperature and pressure) and all the enzymes are highly selective, which allows the reaction to produce three basic chemicals (formic acid, formaldehyde, and methanol) in just one pot. There are various challenges, however, in applying the enzymatic conversion of CO2, namely, to obtain high productivity, increase reusability of the enzymes and cofactors, and to design a simple, facile, and efficient reactor setup that will sustain the multi-enzymatic cascade catalysis. This review reports on enzyme-aided reactor systems that support the reduction of CO2 to methanol. Such systems include enzyme membrane reactors, electrochemical cells, and photocatalytic reactor systems. Existing reactor setups are described, product yields and biocatalytic productivities are evaluated, and effective enzyme immobilization methods are discussed.


2019 ◽  
Vol 7 (1) ◽  
pp. 43-52 ◽  
Author(s):  
Yan Liu ◽  
Yuling Qin ◽  
Yuanlin Zheng ◽  
Yong Qin ◽  
Mengjun Cheng ◽  
...  

A one-pot and modular self-assembly strategy for high-performance enzyme cascade bioplatform based on dual-functionalized protein/inorganic hybrid.


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