scholarly journals One-pot Synthesis of (R)- and (S)-phenylglycinol From Bio-based L-phenylalanine by an Artificial Biocatalytic Cascade

2021 ◽  
Author(s):  
Jiandong Zhang ◽  
Ning Qi ◽  
Lili Gao ◽  
Jing Li ◽  
Chaofeng Zhang ◽  
...  
Keyword(s):  
Glucose Dehydrogenase ◽  
Value Added ◽  
Microbial Consortia ◽  
Acid Decarboxylase ◽  
Grand Challenge ◽  
Preparative Scale ◽  
E Coli ◽  
Ferulic Acid Decarboxylase ◽  
Cascade Biocatalysis ◽  
Cell Module

Abstract Chiral phenylglycinol is a very important chemical in the pharmaceutical manufacturing. Current methods for synthesis of chiral phenylglycinol often suffered from unsatisfied selectivity, low product yield and using the non-renewable resourced substrates, then the synthesis of chiral phenylglycinol remain a grand challenge. Design and construction of synthetic microbial consortia is a promising strategy to convert bio-based materials to high value-added chiral compounds. In this study, we reported a six-step artificial cascade biocatalysis system for conversion of biobased L-phenylalanine to yield chiral phenylglycinol. The cascade biocatalysis system was conducted by a microbial consortium composed of two engineered recombinant Escherichia coli cells modules, one recombinant E. coli cell module co-expression of six different enzymes (phenylalanine ammonia lyase/ferulic acid decarboxylase/phenylacrylic acid decarboxylase/styrene monooxygenase/epoxide hydrolase/alcohol dehydrogenase) for efficient conversion of L-phenylalanine into 2-hydroxyacetophenone. The second recombinant E. coli cell module expression of an (R)-ω-transaminase or co-expression of the (S)-ω-transaminase, alanine dehydrogenase and glucose dehydrogenase for conversion of 2-hydroxyacetophenone to (S)- or (R)-phenylglycinol, respectively. Combining the two engineered E. coli cell modules, after the optimization of bioconversion conditions (including pH, temperature, glucose concentration, amine donor concentration and cell ratio), L-phenylalanine could be easily converted to (R)-phenylglycinol and (S)-phenylglycinol with up to 99% conversion and >99% ee. Preparative scale biotransformation was also conducted on 100 mL scale, (S)-phenylglycinol and (R)-phenylglycinol were obtained in 71.0% and 80.5% yield, >99% ee, and 5.19 g/L.d and 4.42 g/L.d productivity, respectively. The salient features of this biocatalytic cascade system are good yields, excellent ee, mild reaction conditions and no need for additional cofactor (NADH/NAD+), provide a practical biocatalytic method for sustainable synthesis of (S)-phenylglycinol and (R)-phenylglycinol from biobased L-phenylalanine.

scholarly journals One-pot synthesis of (R)- and (S)-phenylglycinol from bio-based l-phenylalanine by an artificial biocatalytic cascade

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Jiandong Zhang ◽  
Ning Qi ◽  
Lili Gao ◽  
Jing Li ◽  
Chaofeng Zhang ◽  
...  
Keyword(s):  
Glucose Dehydrogenase ◽  
Value Added ◽  
Microbial Consortia ◽  
Acid Decarboxylase ◽  
Grand Challenge ◽  
Preparative Scale ◽  
E Coli ◽  
Ferulic Acid Decarboxylase ◽  
Cell Modules ◽  
Cell Module

AbstractChiral phenylglycinol is a very important chemical in the pharmaceutical manufacturing. Current methods for synthesis of chiral phenylglycinol often suffered from unsatisfied selectivity, low product yield and using the non-renewable resourced substrates, then the synthesis of chiral phenylglycinol remain a grand challenge. Design and construction of synthetic microbial consortia is a promising strategy to convert bio-based materials into high value-added chiral compounds. In this study, we reported a six-step artificial cascade biocatalysis system for conversion of bio-based l-phenylalanine into chiral phenylglycinol. This system was designed using a microbial consortium including two engineered recombinant Escherichia coli cell modules, one recombinant E. coli cell module co-expressed six different enzymes (phenylalanine ammonia lyase/ferulic acid decarboxylase/phenylacrylic acid decarboxylase/styrene monooxygenase/epoxide hydrolase/alcohol dehydrogenase) for efficient conversion of l-phenylalanine into 2-hydroxyacetophenone. The second recombinant E. coli cell module expressed an (R)-ω-transaminase or co-expressed the (S)-ω-transaminase, alanine dehydrogenase and glucose dehydrogenase for conversion of 2-hydroxyacetophenone into (S)- or (R)-phenylglycinol, respectively. Combining the two engineered E. coli cell modules, after the optimization of bioconversion conditions (including pH, temperature, glucose concentration, amine donor concentration and cell ratio), l-phenylalanine could be easily converted into (R)-phenylglycinol and (S)-phenylglycinol with up to 99% conversion and > 99% ee. Preparative scale biotransformation was also conducted on 100-mL scale, (S)-phenylglycinol and (R)-phenylglycinol could be obtained in 71.0% and 80.5% yields, > 99% ee, and 5.19 g/L d and 4.42 g/L d productivity, respectively. The salient features of this biocatalytic cascade system are good yields, excellent ee, mild reaction condition and no need for additional cofactor (NADH/NAD+), provide a practical biocatalytic method for sustainable synthesis of (S)-phenylglycinol and (R)-phenylglycinol from bio-based L-phenylalanine.

scholarly journals Refactoring of a synthetic raspberry ketone pathway with EcoFlex

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Simon J. Moore ◽  
Yonek B. Hleba ◽  
Sarah Bischoff ◽  
David Bell ◽  
Karen M. Polizzi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Synthetic Biology ◽  
Combinatorial Libraries ◽  
Value Added ◽  
Microtiter Plate ◽  
Fine Tuning ◽  
Golden Gate ◽  
E Coli ◽  
Fine Chemical ◽  
Raspberry Ketone

Abstract Background  A key focus of synthetic biology is to develop microbial or cell-free based biobased routes to value-added chemicals such as fragrances. Originally, we developed the EcoFlex system, a Golden Gate toolkit, to study genes/pathways flexibly using Escherichia coli heterologous expression. In this current work, we sought to use EcoFlex to optimise a synthetic raspberry ketone biosynthetic pathway. Raspberry ketone is a high-value (~ £20,000 kg−1) fine chemical farmed from raspberry (Rubeus rubrum) fruit. Results  By applying a synthetic biology led design-build-test-learn cycle approach, we refactor the raspberry ketone pathway from a low level of productivity (0.2 mg/L), to achieve a 65-fold (12.9 mg/L) improvement in production. We perform this optimisation at the prototype level (using microtiter plate cultures) with E. coli DH10β, as a routine cloning host. The use of E. coli DH10β facilitates the Golden Gate cloning process for the screening of combinatorial libraries. In addition, we also newly establish a novel colour-based phenotypic screen to identify productive clones quickly from solid/liquid culture. Conclusions  Our findings provide a stable raspberry ketone pathway that relies upon a natural feedstock (L-tyrosine) and uses only constitutive promoters to control gene expression. In conclusion we demonstrate the capability of EcoFlex for fine-tuning a model fine chemical pathway and provide a range of newly characterised promoter tools gene expression in E. coli.

Chemoenzymatic cascade for stilbene production from cinnamic acid catalyzed by ferulic acid decarboxylase and an artificial metathease

2019 ◽  
Vol 9 (20) ◽  
pp. 5572-5576 ◽  
Author(s):  
M. A. Stephanie Mertens ◽  
Daniel F. Sauer ◽  
Ulrich Markel ◽  
Johannes Schiffels ◽  
Jun Okuda ◽  
...  
Keyword(s):  
Ferulic Acid ◽  
Cinnamic Acid ◽  
Olefin Metathesis ◽  
Metal Contamination ◽  
Cascade Reaction ◽  
Acid Decarboxylase ◽  
Efficient Removal ◽  
Ferulic Acid Decarboxylase ◽  
Acid Catalyzed ◽  
Removal Of Metal

We report a chemoenzymatic cascade reaction for stilbene production combining decarboxylation and olefin metathesis with efficient removal of metal contamination.

Mechanistic insights into the catalytic reaction of ferulic acid decarboxylase from Aspergillus niger: a QM/MM study

2017 ◽  
Vol 19 (11) ◽  
pp. 7733-7742 ◽  
Author(s):  
Ge Tian ◽  
Yongjun Liu
Keyword(s):  
Aspergillus Niger ◽  
Ferulic Acid ◽  
Catalytic Reaction ◽  
Acid Decarboxylase ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Ferulic Acid Decarboxylase ◽  
Rate Limiting

QM/MM calculations reveal the cofactor prFMNiminiumto be the catalytically relevant species compared with prFMNketamine. The protonation of the intermediate is the rate-limiting step, and the prolonged leaving of the generated CO2can facilitate this process.

scholarly journals Biological Parts for Plant Biodesign to Enhance Land-Based Carbon Dioxide Removal

2021 ◽  
Vol 2021 ◽  
pp. 1-22
Author(s):  
Xiaohan Yang ◽  
Degao Liu ◽  
Haiwei Lu ◽  
David J. Weston ◽  
Jin-Gui Chen ◽  
...  
Keyword(s):  
Climate Change ◽  
Carbon Dioxide ◽  
Carbon Allocation ◽  
Value Added ◽  
Carbon Dioxide Removal ◽  
Grand Challenge ◽  
Terrestrial Plants ◽  
Long Term Storage ◽  
Term Storage

A grand challenge facing society is climate change caused mainly by rising CO2 concentration in Earth’s atmosphere. Terrestrial plants are linchpins in global carbon cycling, with a unique capability of capturing CO2 via photosynthesis and translocating captured carbon to stems, roots, and soils for long-term storage. However, many researchers postulate that existing land plants cannot meet the ambitious requirement for CO2 removal to mitigate climate change in the future due to low photosynthetic efficiency, limited carbon allocation for long-term storage, and low suitability for the bioeconomy. To address these limitations, there is an urgent need for genetic improvement of existing plants or construction of novel plant systems through biosystems design (or biodesign). Here, we summarize validated biological parts (e.g., protein-encoding genes and noncoding RNAs) for biological engineering of carbon dioxide removal (CDR) traits in terrestrial plants to accelerate land-based decarbonization in bioenergy plantations and agricultural settings and promote a vibrant bioeconomy. Specifically, we first summarize the framework of plant-based CDR (e.g., CO2 capture, translocation, storage, and conversion to value-added products). Then, we highlight some representative biological parts, with experimental evidence, in this framework. Finally, we discuss challenges and strategies for the identification and curation of biological parts for CDR engineering in plants.

scholarly journals Efficient production of (S)-1-phenyl-1, 2-ethanediol using xylan as co-substrate by a coupled multi-enzyme Escherichia coli system

2020 ◽  
Author(s):  
Junchao Rao ◽  
Rongzhen Zhang ◽  
Guanyu Xu ◽  
Lihong Li ◽  
Yan Xu
Keyword(s):  
Escherichia Coli ◽  
Glucose Dehydrogenase ◽  
Optical Purity ◽  
Carbonyl Reductase ◽  
Cofactor Regeneration ◽  
Efficient Production ◽  
Chiral Synthesis ◽  
E Coli ◽  
Concomitant Reduction ◽  
Different Strains

Abstract Background: ( S )-1-phenyl-1,2-ethanediol is an important chiral intermediate in the synthesis of liquid crystals and chiral biphosphines.(S)-carbonyl reductase II from Candida parapsilosis catalyzes the conversion of 2-hydroxyacetophenone to ( S )-1-phenyl-1,2-ethanediol with NADPH as a cofactor. Glucose dehydrogenase with a Ala258Phe mutation is able to catalyze the oxidation of xylose with concomitant reduction of NADP + to NADPH, while endo-β-1,4-xylanase 2 catalyzes the conversion of xylan to xylose. In the present work, the Ala258Phe glucose dehydrogenase mutant and endo-β-1,4-xylanase 2 were introduced into the ( S )-carbonyl reductase II-mediated chiral pathway to strengthen cofactor regeneration by using xylan as a naturally abundant co-substrate. Results: We constructed several coupled multi-enzyme systems by introducing ( S )-carbonyl reductase II, the A258F glucose dehydrogenase mutant and endo-β-1,4-xylanase 2 into Escherichia coli . Different strains were produced by altering the location of the encoding genes on the plasmid. Only recombinant E. coli /pET-G-S-2 expressed all three enzymes, and this strain produced ( S )-1-phenyl-1,2-ethanediol from 2-hydroxyacetophenone as a substrate and xylan as a co-substrate. The optical purity was 100% and the yield was 98.3% (6 g/L 2-HAP) under optimal conditions of 35°C, pH 6.5 and a 2:1 substrate-co-substrate ratio. The introduction of A258F glucose dehydrogenase and endo-β-1,4-xylanase 2 into the ( S )-carbonyl reductase II-mediated chiral pathway caused a 54.6% increase in yield, and simultaneously reduced the reaction time from 48 h to 28 h. Conclusions: This study demonstrates efficient chiral synthesis using a pentose as a co-substrate to enhance cofactor regeneration. This provides a new approach for enantiomeric catalysis through the inclusion of naturally abundant materials.

Cascaded Biocatalysis and Bioelectrocatalysis: Overview and Recent Advances

2021 ◽  
Vol 72 (1) ◽  
Author(s):  
Yoo Seok Lee ◽  
Koun Lim ◽  
Shelley D. Minteer
Keyword(s):  
Value Added ◽  
Cascade Reactions ◽  
Product Formation ◽  
Deep Oxidation ◽  
Annual Review ◽  
Publication Date ◽  
Oxidation Reduction ◽  
Cascade Biocatalysis ◽  
Analyte Detection ◽  
Catalyst Systems

Enzyme cascades are plentiful in nature, but they also have potential in artificial applications due to the possibility of using the target substrate in biofuel cells, electrosynthesis, and biosensors. Cascade reactions from enzymes or hybrid bioorganic catalyst systems exhibit extended substrate range, reaction depth, and increased overall performance. This review addresses the strategies of cascade biocatalysis and bioelectrocatalysis for ( a) CO2 fixation, ( b) high value-added product formation, ( c) sustainable energy sources via deep oxidation, and ( d) cascaded electrochemical enzymatic biosensors. These recent updates in the field provide fundamental concepts, designs of artificial electrocatalytic oxidation-reduction pathways (using a flexible setup involving organic catalysts and engineered enzymes), and advances in hybrid cascaded sensors for sensitive analyte detection. Expected final online publication date for the Annual Review of Physical Chemistry, Volume 72 is April 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

scholarly journals Structural Basis of Enzymatic Activity for the Ferulic Acid Decarboxylase (FADase) from Enterobacter sp. Px6-4

PLoS ONE ◽  
2011 ◽  
Vol 6 (1) ◽  
pp. e16262 ◽  
Author(s):  
Wen Gu ◽  
Jinkui Yang ◽  
Zhiyong Lou ◽  
Lianming Liang ◽  
Yuna Sun ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Ferulic Acid ◽  
Structural Basis ◽  
Acid Decarboxylase ◽  
Ferulic Acid Decarboxylase

scholarly journals Upcycling chitin-containing waste into organonitrogen chemicals via an integrated process

2020 ◽  
Vol 117 (14) ◽  
pp. 7719-7728 ◽  
Author(s):  
Xiaoqiang Ma ◽  
Gökalp Gözaydın ◽  
Huiying Yang ◽  
Wenbo Ning ◽  
Xi Han ◽  
...  
Keyword(s):  
Wide Spectrum ◽  
Value Added ◽  
Direct Conversion ◽  
Water Soluble ◽  
Integrated Biorefinery ◽  
E Coli ◽  
Shrimp Shell ◽  
Shell Waste ◽  
Shrimp Shell Waste ◽  
Value Added Products

Chitin is the most abundant renewable nitrogenous material on earth and is accessible to humans in the form of crustacean shell waste. Such waste has been severely underutilized, resulting in both resource wastage and disposal issues. Upcycling chitin-containing waste into value-added products is an attractive solution. However, the direct conversion of crustacean shell waste-derived chitin into a wide spectrum of nitrogen-containing chemicals (NCCs) is challenging via conventional catalytic processes. To address this challenge, in this study, we developed an integrated biorefinery process to upgrade shell waste-derived chitin into two aromatic NCCs that currently cannot be synthesized from chitin via any chemical process (tyrosine andl-DOPA). The process involves a pretreatment of chitin-containing shell waste and an enzymatic/fermentative bioprocess using metabolically engineeredEscherichia coli. The pretreatment step achieved an almost 100% recovery and partial depolymerization of chitin from shrimp shell waste (SSW), thereby offering water-soluble chitin hydrolysates for the downstream microbial process under mild conditions. The engineeredE. colistrains produced 0.91 g/L tyrosine or 0.41 g/Ll-DOPA from 22.5 g/L unpurified SSW-derived chitin hydrolysates, demonstrating the feasibility of upcycling renewable chitin-containing waste into value-added NCCs via this integrated biorefinery, which bypassed the Haber–Bosch process in providing a nitrogen source.

Review on biological wastewater treatment and resources recovery: attached and suspended growth systems

2019 ◽  
Vol 80 (11) ◽  
pp. 2013-2026 ◽  
Author(s):  
Lakshmi Machineni
Keyword(s):  
Wastewater Treatment ◽  
Biological Treatment ◽  
Film Growth ◽  
Value Added ◽  
Microbial Consortia ◽  
Biological Wastewater Treatment ◽  
Suspended Growth ◽  
Growth System ◽  
Pure Cultures ◽  
Biological Methods

Abstract The treatment of wastewater for reuse is a potential solution to meet ever increasing urban, industrial, agricultural, and environmental demands across the world, where clean water availability is scarce. There are several traditional wastewater treatment processes that offer varying degrees of effectiveness in addition to presenting environmental, economic, and social disadvantages. Development of promising and inexpensive technologies to provide the reusable water in needful amounts using wastewaters as a cheap source of key nutrients and organic matter is required. Wastewater treatment by biological methods is becoming more important in the light of recovering value-added plant nutrients, heavy metals, biosolids, and bioenergy resources. Different types of solid contaminants in effluents can be removed simultaneously by pure cultures or mixed microbial consortia. Based on the structural organization of microbial biomass, biological treatment systems are classified into two types: dispersed growth system and attached growth system. Biological treatment methods associated with fixed-film growth have been recognized as highly effective and more energy efficient than suspended growth systems. This review discusses the recent breakthroughs in advanced biological wastewater treatment using both the systems, and also focuses on key energetic resources recovery driven by biological technologies.

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