Bioproduction of Benzylamine from Renewable Feedstocks via a Nine-Step Artificial Enzyme Cascade and Engineered Metabolic Pathways

ChemSusChem ◽  
2018 ◽  
Vol 11 (13) ◽  
pp. 2221-2228 ◽  
Author(s):  
Yi Zhou ◽  
Shuke Wu ◽  
Jiwei Mao ◽  
Zhi Li
2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Benedict Ryan Lukito ◽  
Zilong Wang ◽  
Balaji Sundara Sekar ◽  
Zhi Li

Abstract(R)-mandelic acid is an industrially important chemical, especially used for producing antibiotics. Its chemical synthesis often uses highly toxic cyanide to produce its racemic form, followed by kinetic resolution with 50% maximum yield. Here we report a green and sustainable biocatalytic method for producing (R)-mandelic acid from easily available styrene, biobased L-phenylalanine, and renewable feedstocks such as glycerol and glucose, respectively. An epoxidation-hydrolysis-double oxidation artificial enzyme cascade was developed to produce (R)-mandelic acid at 1.52 g/L from styrene with > 99% ee. Incorporation of deamination and decarboxylation into the above cascade enables direct conversion of L-phenylalanine to (R)-mandelic acid at 913 mg/L and > 99% ee. Expressing the five-enzyme cascade in an L-phenylalanine-overproducing E. coli NST74 strain led to the direct synthesis of (R)-mandelic acid from glycerol or glucose, affording 228 or 152 mg/L product via fermentation. Moreover, coupling of E. coli cells expressing L-phenylalanine biosynthesis pathway with E. coli cells expressing the artificial enzyme cascade enabled the production of 760 or 455 mg/L (R)-mandelic acid from glycerol or glucose. These simple, safe, and green methods show great potential in producing (R)-mandelic acid from renewable feedstocks.


2021 ◽  
Author(s):  
Boyu Yang ◽  
Shubin Li ◽  
Wei Mu ◽  
Zhao Wang ◽  
Xiaojun Han

AbstractThe bottom-up constructed artificial cells help to understand the cell working mechanism and provide the evolution clues for organisms. Cyanobacteria are believed to be the ancestors of chloroplasts according to endosymbiosis theory. Herein we demonstrate an artificial cell containing cyanobacteria to mimic endosymbiosis phenomenon. The cyanobacteria sustainably produce glucose molecules by converting light energy into chemical energy. Two downstream “metabolic” pathways starting from glucose molecules are investigated. One involves enzyme cascade reaction to produce H2O2 (assisted by glucose oxidase) first, followed by converting Amplex red to resorufin (assisted by horseradish peroxidase). The more biological one involves nicotinamide adenine dinucleotide (NADH) production in the presence of NAD+ and glucose dehydrogenase. Further, NADH molecules are oxidized into NAD+ by pyruvate catalyzed by lactate dehydrogenase, meanwhile, lactate is obtained. Therefore, the sustainable cascade cycling of NADH/NAD+ is built. The artificial cells built here simulate the endosymbiosis phenomenon, meanwhile pave the way for investigating more complicated sustainable energy supplied metabolism inside artificial cells.


2019 ◽  
Vol 41 (4-5) ◽  
pp. 605-611 ◽  
Author(s):  
Takenori Satomura ◽  
Kousaku Horinaga ◽  
Shino Tanaka ◽  
Eiichiro Takamura ◽  
Hiroaki Sakamoto ◽  
...  

2015 ◽  
Vol 71 (12) ◽  
pp. 1475-1480
Author(s):  
Iuliia Iermak ◽  
Oksana Degtjarik ◽  
Fabian Steffler ◽  
Volker Sieber ◽  
Ivana Kuta Smatanova

The glyceraldehyde dehydrogenase fromThermoplasma acidophilum(TaAlDH) is a microbial enzyme that catalyzes the oxidation of D-glyceraldehyde to D-glycerate in the artificial enzyme cascade designed for the conversion of glucose to the organic solvents isobutanol and ethanol. Various mutants ofTaAlDH were constructed by a random approach followed by site-directed and saturation mutagenesis in order to improve the properties of the enzyme that are essential for its functioning within the cascade. Two enzyme variants, wild-typeTaAlDH (TaAlDHwt) and an F34M+S405N variant (TaAlDH F34M+S405N), were successfully crystallized. Crystals ofTaAlDHwt belonged to the monoclinic space groupP1211 with eight molecules per asymmetric unit and diffracted to a resolution of 1.95 Å.TaAlDH F34M+S405N crystallized in two different space groups: triclinicP1 with 16 molecules per asymmetric unit and monoclinicC121 with four molecules per asymmetric unit. These crystals diffracted to resolutions of 2.14 and 2.10 Å for theP1 andC121 crystals, respectively.


Author(s):  
Marlene Pröschel ◽  
Rainer Detsch ◽  
Aldo R. Boccaccini ◽  
Uwe Sonnewald

Talanta ◽  
2020 ◽  
Vol 220 ◽  
pp. 121374 ◽  
Author(s):  
Min Liu ◽  
Junsong Mou ◽  
Xiaohan Xu ◽  
Feifei Zhang ◽  
Jianfei Xia ◽  
...  

2014 ◽  
Vol 31 ◽  
pp. S75
Author(s):  
Wolfgang Kroutil ◽  
Johann Sattler ◽  
Michael Fuchs ◽  
Verena Resch ◽  
Joerg Schrittwieser

ChemCatChem ◽  
2015 ◽  
Vol 7 (23) ◽  
pp. 3951-3955 ◽  
Author(s):  
Sandy Schmidt ◽  
Hanna C. Büchsenschütz ◽  
Christian Scherkus ◽  
Andreas Liese ◽  
Harald Gröger ◽  
...  

The Analyst ◽  
2021 ◽  
Author(s):  
Huiying Xu ◽  
Lu Zheng ◽  
Yu Zhou ◽  
Bang-Ce Ye

Tumor-related exosomes, which are heterogeneous membrane-enclosed nanovesicles shed from cancer cells, have been widely recognized as potential noninvasive biomarkers for early cancer diagnosis. Herein, an artificial enzyme cascade amplification strategy...


Author(s):  
Yilan Liu ◽  
Mauricio Garcia Benitez ◽  
Jinjin Chen ◽  
Emma Harrison ◽  
Anna N. Khusnutdinova ◽  
...  

Global warming and uneven distribution of fossil fuels worldwide concerns have spurred the development of alternative, renewable, sustainable, and environmentally friendly resources. From an engineering perspective, biosynthesis of fatty acid-derived chemicals (FACs) is an attractive and promising solution to produce chemicals from abundant renewable feedstocks and carbon dioxide in microbial chassis. However, several factors limit the viability of this process. This review first summarizes the types of FACs and their widely applications. Next, we take a deep look into the microbial platform to produce FACs, give an outlook for the platform development. Then we discuss the bottlenecks in metabolic pathways and supply possible solutions correspondingly. Finally, we highlight the most recent advances in the fast-growing model-based strain design for FACs biosynthesis.


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