Production of Optically Pure Aryl Epoxides by Recombinant E. coli Carrying Styrene Monooxygenase

2001 ◽  
pp. 141-149
Author(s):  
A. Colmegna ◽  
E. Galli ◽  
G. Sello ◽  
G. Bestetti
Keyword(s):  
E Coli ◽  
Styrene Monooxygenase ◽  
Optically Pure

scholarly journals Production of Enantiopure Chiral Epoxides with E. coli Expressing Styrene Monooxygenase

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1514
Author(s):  
Dominika Gyuranová ◽  
Radka Štadániová ◽  
Zuzana Hegyi ◽  
Róbert Fischer ◽  
Martin Rebroš
Keyword(s):  
High Cell Density ◽  
Enantiomeric Excess ◽  
Building Blocks ◽  
E Coli ◽  
Styrene Monooxygenase ◽  
Chiral Compounds ◽  
Chiral Epoxides ◽  
Optically Pure ◽  
Epoxidation Of Alkenes ◽  
Excellent Stability

Styrene monooxygenases are a group of highly selective enzymes able to catalyse the epoxidation of alkenes to corresponding chiral epoxides in excellent enantiopurity. Chiral compounds containing oxirane ring or products of their hydrolysis represent key building blocks and precursors in organic synthesis in the pharmaceutical industry, and many of them are produced on an industrial scale. Two-component recombinant styrene monooxygenase (SMO) from Marinobacterium litorale was expressed as a fused protein (StyAL2StyB) in Escherichia coli BL21(DE3). By high cell density fermentation, 35 gDCW/L of biomass with overexpressed SMO was produced. SMO exhibited excellent stability, broad substrate specificity, and enantioselectivity, as it remained active for months and converted a group of alkenes to corresponding chiral epoxides in high enantiomeric excess (˃95–99% ee). Optically pure (S)-4-chlorostyrene oxide, (S)-allylbenzene oxide, (2R,5R)-1,2:5,6-diepoxyhexane, 2-(3-bromopropyl)oxirane, and (S)-4-(oxiran-2-yl)butan-1-ol were prepared by whole-cell SMO.

Scalable biocatalytic synthesis of optically pure ethyl (R)-2-hydroxy-4-phenylbutyrate using a recombinant E. coli with high catalyst yield

2013 ◽  
Vol 168 (4) ◽  
pp. 493-498 ◽  
Author(s):  
Ye Ni ◽  
Yuning Su ◽  
Haidong Li ◽  
Jieyu Zhou ◽  
Zhihao Sun
Keyword(s):  
E Coli ◽  
Optically Pure

A New Biocatalyst for Production of Optically Pure Aryl Epoxides by Styrene Monooxygenase from Pseudomonas fluorescensST

1999 ◽  
Vol 65 (6) ◽  
pp. 2794-2797 ◽  
Author(s):  
Patrizia Di Gennaro ◽  
Andrea Colmegna ◽  
Enrica Galli ◽  
Guido Sello ◽  
Francesca Pelizzoni ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Fluorescens ◽  
Substrate Preference ◽  
Fine Chemicals ◽  
Styrene Monooxygenase ◽  
Optically Pure

ABSTRACT We developed a biocatalyst by cloning the styrene monooxygenase genes (styA and styB) from Pseudomonas fluorescens ST responsible for the oxidation of styrene to its corresponding epoxide. Recombinant Escherichia coli was able to oxidize different aryl vinyl and aryl ethenyl compounds to their corresponding optically pure epoxides. The results of bioconversions indicate the broad substrate preference of styrene monooxygenase and its potential for the production of several fine chemicals.

A synthetic biology approach for the transformation of l-α-amino acids to the corresponding enantiopure (R)- or (S)-α-hydroxy acids

2015 ◽  
Vol 51 (14) ◽  
pp. 2828-2831 ◽  
Author(s):  
Geoffrey Gourinchas ◽  
Eduardo Busto ◽  
Manuela Killinger ◽  
Nina Richter ◽  
Birgit Wiltschi ◽  
...  
Keyword(s):  
Amino Acids ◽  
Synthetic Biology ◽  
Hydroxy Acids ◽  
E Coli ◽  
Optically Pure ◽  
Synthetic Biology Approach

A synthetic biology approach enabled the design of a single E. coli cell catalyst co-expressing three enzymes (l-AAD, l- or d-HIC and FDH) for the quantitatively transformation of l-amino acids to the corresponding optically pure (R)- and (S)-α-hydroxy acids.

Homofermentative Production of d- orl-Lactate in Metabolically Engineered Escherichia coli RR1

1999 ◽  
Vol 65 (4) ◽  
pp. 1384-1389 ◽  
Author(s):  
Dong-Eun Chang ◽  
Heung-Chae Jung ◽  
Joon-Shick Rhee ◽  
Jae-Gu Pan
Keyword(s):  
Escherichia Coli ◽  
Lactate Dehydrogenase ◽  
Lactobacillus Casei ◽  
Lactate Production ◽  
Anaerobic Conditions ◽  
E Coli ◽  
Fermentation Product ◽  
Dehydrogenase Gene ◽  
Mutant Strains ◽  
Optically Pure

ABSTRACT We investigated metabolic engineering of fermentation pathways inEscherichia coli for production of optically pured- or l-lactate. Several pta mutant strains were examined, and a pta mutant of E. coli RR1 which was deficient in the phosphotransacetylase of the Pta-AckA pathway was found to metabolize glucose tod-lactate and to produce a small amount of succinate by-product under anaerobic conditions. An additional mutation inppc made the mutant produce d-lactate like a homofermentative lactic acid bacterium. When the pta ppcdouble mutant was grown to higher biomass concentrations under aerobic conditions before it shifted to the anaerobic phase ofd-lactate production, more than 62.2 g ofd-lactate per liter was produced in 60 h, and the volumetric productivity was 1.04 g/liter/h. To examine whether the blocked acetate flux could be reoriented to a nonindigenousl-lactate pathway, an l-lactate dehydrogenase gene from Lactobacillus casei was introduced into apta ldhA strain which lacked phosphotransacetylase andd-lactate dehydrogenase. This recombinant strain was able to metabolize glucose to l-lactate as the major fermentation product, and up to 45 g of l-lactate per liter was produced in 67 h. These results demonstrate that the central fermentation metabolism of E. coli can be reoriented to the production of d-lactate, an indigenous fermentation product, or to the production of l-lactate, a nonindigenous fermentation product.

Baeyer-Villiger oxidations catalyzed by engineered microorganisms: Enantioselective synthesis of δ-valerolactones with functionalized chains

2002 ◽  
Vol 80 (6) ◽  
pp. 613-621 ◽  
Author(s):  
Shaozhao Wang ◽  
Gang Chen ◽  
Margaret M Kayser ◽  
Hiroaki Iwaki ◽  
Peter C.K Lau ◽  
...  
Keyword(s):  
Baker’S Yeast ◽  
Enantioselective Synthesis ◽  
Baker's Yeast ◽  
Pseudomonas Sp ◽  
Cyclohexanone Monooxygenase ◽  
E Coli ◽  
Acinetobacter Sp ◽  
Optically Pure

Cyclohexanone monooxygenase (CHMO) from Acinetobacter sp NCIMB 9871 expressed in baker's yeast and in E. coli and cyclopentanone monooxygenase (CPMO) from Comamonas (previously Pseudomonas) sp. NCIMB 9872 expressed in E. coli are new bioreagents for Baeyer-Villiger oxidations. These engineered microorganisms, requiring neither biochemical expertise nor equipment beyond that found in chemical laboratories, were evaluated as reagents for Baeyer-Villiger oxidations of cyclopentanones substituted at the 2-position with polar and nonpolar chains suitable for further modifications. Two such functionalized substrates that can be transformed into highly enantiopure lactones were identified. The performance and the potential of these bioreagents are discussed.Key words: enantioselective Baeyer-Villiger oxidations, biotransformations, cyclohexanone monooxygenase, cyclo pentanone monooxygenase, engineered baker's yeast, recombinant E. coli, optically pure 2-substituted cyclopentanones, optically pure lactones.

scholarly journals Purification and Gene Cloning of α-Methylserine Aldolase from Ralstonia sp. Strain AJ110405 and Application of the Enzyme in the Synthesis of α-Methyl-l-Serine

2008 ◽  
Vol 74 (24) ◽  
pp. 7596-7599 ◽  
Author(s):  
Hiroyuki Nozaki ◽  
Shinji Kuroda ◽  
Kunihiko Watanabe ◽  
Kenzo Yokozeki
Keyword(s):  
Escherichia Coli ◽  
Enzymatic Activity ◽  
High Yield ◽  
Synthesis Reaction ◽  
Gene Encoding ◽  
E Coli ◽  
Variovorax Paradoxus ◽  
Excess Amount ◽  
Synthesis Activity ◽  
Optically Pure

ABSTRACT By screening microorganisms that are capable of assimilating α-methyl-dl-serine, we detected α-methylserine aldolase in Ralstonia sp. strain AJ110405, Variovorax paradoxus AJ110406, and Bosea sp. strain AJ110407. A homogeneous form of this enzyme was purified from Ralstonia sp. strain AJ110405, and the gene encoding the enzyme was cloned and expressed in Escherichia coli. The enzyme appeared to be a homodimer consisting of identical subunits, and its molecular mass was found to be 47 kDa. It contained 0.7 to 0.8 mol of pyridoxal 5′-phosphate per mol of subunit and could catalyze the interconversion of α-methyl-l-serine to l-alanine and formaldehyde in the absence of tetrahydrofolate. Formaldehyde was generated from α-methyl-l-serine but not from α-methyl-d-serine, l-serine, or d-serine. α-Methyl-l-serine synthesis activity was detected when l-alanine was used as the substrate. In contrast, no activity was detected when d-alanine was used as the substrate. In the α-methyl-l-serine synthesis reaction, the enzymatic activity was inhibited by an excess amount of formaldehyde, which was one of the substrates. We used cells of E. coli as a whole-cell catalyst to express the gene encoding α-methylserine aldolase and effectively obtained a high yield of optically pure α-methyl-l-serine using l-alanine and formaldehyde.

Endotoxin Alteration of the Mucopolysaccharide Blood Vessel Coating in Rats

1974 ◽  
Vol 32 ◽  
pp. 148-149
Author(s):  
D. E. Philpott ◽  
A. Takahashi
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Salivary Gland ◽  
Carotid Artery ◽  
Basement Membrane ◽  
Coronary Vessels ◽  
Ruthenium Red ◽  
E Coli ◽  
Old Rats ◽  
The Brain

Two month, eight month and two year old rats were treated with 10 or 20 mg/kg of E. Coli endotoxin I. P. The eight month old rats proved most resistant to the endotoxin. During fixation the aorta, carotid artery, basil arartery of the brain, coronary vessels of the heart, inner surfaces of the heart chambers, heart and skeletal muscle, lung, liver, kidney, spleen, brain, retina, trachae, intestine, salivary gland, adrenal gland and gingiva were treated with ruthenium red or alcian blue to preserve the mucopolysaccharide (MPS) coating. Five, 8 and 24 hrs of endotoxin treatment produced increasingly marked capillary damage, disappearance of the MPS coating, edema, destruction of endothelial cells and damage to the basement membrane in the liver, kidney and lung.

Ribosome Structural Organization

1974 ◽  
Vol 32 ◽  
pp. 332-333
Author(s):  
James A. Lake
Keyword(s):  
Ribosomal Proteins ◽  
Structural Organization ◽  
Three Dimensional ◽  
Small Subunit ◽  
Structural Studies ◽  
X Ray Diffraction ◽  
Specific Antibodies ◽  
X Ray ◽  
E Coli ◽  
Complete Sequences

The understanding of ribosome structure has advanced considerably in the last several years. Biochemists have characterized the constituent proteins and rRNA's of ribosomes. Complete sequences have been determined for some ribosomal proteins and specific antibodies have been prepared against all E. coli small subunit proteins. In addition, a number of naturally occuring systems of three dimensional ribosome crystals which are suitable for structural studies have been observed in eukaryotes. Although the crystals are, in general, too small for X-ray diffraction, their size is ideal for electron microscopy.

Sites of Adsorption of the Bacteriophages T3 and T5 on the Wall of Escherichia Coli B.

1967 ◽  
Vol 25 ◽  
pp. 96-97
Author(s):  
Manfred E. Bayer
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Electron Microscope ◽  
Uranyl Acetate ◽  
E Coli ◽  
Receptor Sites ◽  
Rigid Cell Wall ◽  
Host Cell Surface ◽  
Bacterial Viruses ◽  
Escherichia Coli B

Bacterial viruses adsorb specifically to receptors on the host cell surface. Although the chemical composition of some of the cell wall receptors for bacteriophages of the T-series has been described and the number of receptor sites has been estimated to be 150 to 300 per E. coli cell, the localization of the sites on the bacterial wall has been unknown.When logarithmically growing cells of E. coli are transferred into a medium containing 20% sucrose, the cells plasmolize: the protoplast shrinks and becomes separated from the somewhat rigid cell wall. When these cells are fixed in 8% Formaldehyde, post-fixed in OsO4/uranyl acetate, embedded in Vestopal W, then cut in an ultramicrotome and observed with the electron microscope, the separation of protoplast and wall becomes clearly visible, (Fig. 1, 2). At a number of locations however, the protoplasmic membrane adheres to the wall even under the considerable pull of the shrinking protoplast. Thus numerous connecting bridges are maintained between protoplast and cell wall. Estimations of the total number of such wall/membrane associations yield a number of about 300 per cell.

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