Choice of biocatalyst form for scalable processes

2006 ◽  
Vol 34 (2) ◽  
pp. 301-303 ◽  
Author(s):  
J.M. Woodley
Keyword(s):  
Protein Engineering ◽  
Process Engineering ◽  
Synthetic Chemistry ◽  
Whole Cell ◽  
Recent Developments

The design of biocatalytic processes for industrial synthetic chemistry is determined in large part by the choice of isolated enzyme or whole-cell catalyst form. In the present paper, the considerations for choice are identified and some important classes of bioconversion are discussed in relation to the choice to be made. Recent developments in cell and protein engineering as well as reactor and process engineering are discussed in addition.

Recent developments in the synthetic chemistry of technetium disulfide

2013 ◽  
Vol 42 (44) ◽  
pp. 15540 ◽  
Author(s):  
Maryline Ferrier ◽  
William M. Kerlin ◽  
Frederic Poineau ◽  
Alfred P. Sattelberger ◽  
Kenneth R. Czerwinski
Keyword(s):  
Synthetic Chemistry ◽  
Recent Developments

scholarly journals Rational Protein Engineering of Bacterial N-Demethylases to Create Biocatalysts for the Production of Methylxanthines

2021 ◽  
Author(s):  
Shelby Brooks Mills ◽  
Meredith B Mock ◽  
Ryan M Summers
Keyword(s):  
Protein Engineering ◽  
Pseudomonas Putida ◽  
Point Mutations ◽  
Attractive Alternative ◽  
Methyl Groups ◽  
Specific Point ◽  
Whole Cell ◽  
Methyl Group ◽  
E Coli ◽  
Rich History

Methylxanthines have a rich history as therapeutics and pharmaceuticals. However, natural dimethyl- and monomethylxanthines are difficult to produce synthetically, which has limited further exploration of these compounds in medicinal applications. A biosynthetic method for production of methylxanthines from whole cell biocatalysts is an attractive alternative. The bacterium Pseudomonas putida CBB5 contains a set of five enzymes, NdmABCDE, which are responsible for methylxanthine metabolism via N-demethylation to xanthine. The recent elucidation of the crystal structures of NdmA and NdmB, which remove the N1- and N3- methyl groups of caffeine, respectively, has opened new avenues to create biocatalysts for methylxanthine production. We have created a set of fifteen N-demethylase mutants and expressed them in E. coli BL21(DE3) as whole cell biocatalysts. The activity of each mutant was characterized for their affinity towards caffeine, theobromine, and theophylline. Two mutant enzymes in particular, labeled NdmA3 and NdmA4, both exhibited selectivity towards the N3-methyl group instead of the N1-methyl group. We also discovered that specific point mutations in NdmD resulted in the ability to tune the rate of the N-demethylase reaction. These new enzymes provide the capability of producing high-value methylxanthines, such as paraxanthine and 1-methylxanthine, through a biocatalytic route.

scholarly journals Enhanced Production of (S)-2-arylpropionic Acids by Protein Engineering and Whole-Cell Catalysis

2021 ◽  
Vol 9 ◽  
Author(s):  
Xiaolong Liu ◽  
Meng Zhao ◽  
Xinjiong Fan ◽  
Yao Fu
Keyword(s):  
Protein Engineering ◽  
Ethyl Ester ◽  
Rational Design ◽  
Kinetic Resolution ◽  
Structural Information ◽  
Careful Analysis ◽  
Whole Cell ◽  
Enhanced Production ◽  
Thaw Cycle ◽  
Freeze Thaw Cycle

Esterases are important biocatalysts for chemical synthesis. Several bHSL family esterases have been used to prepare (S)-2-arylpropionic acids with stronger anti-inflammatory effects via kinetic resolution. Here, we presented the discovery of key residues that controlled the enantioselectivity of bHSL family esterases to ethyl 2-arylpropionates, through careful analysis of the structural information and molecular docking. A new bHSL family esterase, Est924, was identified as a promising catalyst for kinetic resolution of racemic ethyl 2-arylpropionates with slight (R)-stereopreference. Using Est924 as the starting enzyme, protein engineering was conducted at hotspots, and the substitution of A203 was proved to enhance the enantioselectivity. The stereopreference of the mutant M1 (A203W) was inverted to ethyl (S)-2-arylpropionates, and this stereopreference was further improved in variant M3 (I202F/A203W/G208F). In addition, the optimal variant, M3, was also suitable for the resolution of ibuprofen ethyl ester and ketoprofen ethyl ester, and their efficient (S)-isomers were synthesized. Next, the whole-cell catalyst harboring M3 was used to prepare (S)-ketoprofen. (S)-ketoprofen with 86%ee was produced by whole-cell catalyst with a single freeze-thaw cycle, and the cells could be reused for at least five cycles. Our results suggested that Est924 variants could kinetically resolve economically important racemates for industrial production and further offer the opportunity for the rational design of enzyme enantioselectivity. Moreover, it is an economical process to prepare optically pure (S)-ketoprofen and (S)-naproxen by using an engineered strain harboring M3 as the catalyst.

scholarly journals Coding Synthetic Chemistry Strategies into Bacterial Designer Cells

2021 ◽  
Author(s):  
Yu-Chang Liu ◽  
Zhong-Liu Wu ◽  
Jan Deska
Keyword(s):  
Building Blocks ◽  
Cell System ◽  
Synthetic Chemistry ◽  
Bacterial Host ◽  
Whole Cell ◽  
Isolation And Purification ◽  
Platform Chemicals ◽  
Natural Enzyme ◽  
High Degree

<p>Following a synthetic chemistry blueprint for the valorization of lignocellulosic platform chemicals, this study showcases a so far unprecedented approach to implement non-natural enzyme modules in vivo. For the design of a novel functional whole cell tool, two purely abiotic transformations were incorporated into a recombinant bacterial host that allows production of complex lactone building blocks. This whole cell system streamlines the synthetic cascade, eliminates isolation and purification steps, and provides a high degree stereoselectivity that has so far been elusive in the chemical methodology.</p>

scholarly journals Coding Synthetic Chemistry Strategies into Bacterial Designer Cells

2021 ◽  
Author(s):  
Yu-Chang Liu ◽  
Zhong-Liu Wu ◽  
Jan Deska
Keyword(s):  
Building Blocks ◽  
Cell System ◽  
Synthetic Chemistry ◽  
Bacterial Host ◽  
Whole Cell ◽  
Isolation And Purification ◽  
Platform Chemicals ◽  
Natural Enzyme ◽  
High Degree

<p>Following a synthetic chemistry blueprint for the valorization of lignocellulosic platform chemicals, this study showcases a so far unprecedented approach to implement non-natural enzyme modules in vivo. For the design of a novel functional whole cell tool, two purely abiotic transformations were incorporated into a recombinant bacterial host that allows production of complex lactone building blocks. This whole cell system streamlines the synthetic cascade, eliminates isolation and purification steps, and provides a high degree stereoselectivity that has so far been elusive in the chemical methodology.</p>

Positron imaging techniques for process engineering: recent developments at Birmingham

2008 ◽  
Vol 19 (9) ◽  
pp. 094004 ◽  
Author(s):  
D J Parker ◽  
T W Leadbeater ◽  
X Fan ◽  
M N Hausard ◽  
A Ingram ◽  
...  
Keyword(s):  
Imaging Techniques ◽  
Process Engineering ◽  
Recent Developments

scholarly journals Biological Insights into Therapeutic Protein Modifications throughout Trafficking and Their Biopharmaceutical Applications

2013 ◽  
Vol 2013 ◽  
pp. 1-19 ◽  
Author(s):  
Xiaotian Zhong ◽  
Jill F. Wright
Keyword(s):  
Protein Engineering ◽  
Cell Line ◽  
Molecular Mechanisms ◽  
Functional Relationship ◽  
Process Engineering ◽  
Cellular Processes ◽  
Therapeutic Properties ◽  
Underlying Mechanisms ◽  
Line Engineering ◽  
Multiple Categories

Over the lifespan of therapeutic proteins, from the point of biosynthesis to the complete clearance from tested subjects, they undergo various biological modifications. Therapeutic influences and molecular mechanisms of these modifications have been well appreciated for some while remained less understood for many. This paper has classified these modifications into multiple categories, according to their processing locations and enzymatic involvement during the trafficking events. It also focuses on the underlying mechanisms and structural-functional relationship between modifications and therapeutic properties. In addition, recent advances in protein engineering, cell line engineering, and process engineering, by exploring these complex cellular processes, are discussed and summarized, for improving functional characteristics and attributes of protein-based biopharmaceutical products.

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