Biotransformation of ferulic acid to protocatechuic acid by Corynebacterium glutamicum ATCC 21420 engineered to express vanillate O-demethylase.

Ferulic acid (4-hydroxy-3-methoxycinnamic acid, FA) is a lignin-derived phenolic compound abundant in plant biomass. The utilization of FA and its conversion to valuable compounds is desired. Protocatechuic acid (3,4-dihydroxybenzoic acid, PCA) is a precursor of polymers and plastics and a constituent of food. A microbial conversion system to produce PCA from FA was developed in this study using a PCA-producing strain of Corynebacterium glutamicum F (ATCC 21420). C. glutamicum strain F grown at 30 °C for 48 h utilized 2 mM each of FA and vanillic acid (4-hydroxy-3-methoxybenzoic acid, VA) to produce PCA, which was secreted into the medium. FA may be catabolized by C. glutamicum through proposed (I) non-ß-oxidative, CoA-dependent or (II) ß-oxidative, CoA-dependent phenylpropanoid pathways. The conversion of VA to PCA is the last step in each pathway. Therefore, the vanillate O-demethylase gene (vanAB) from Corynebacterium efficiens NBRC 100395 was expressed in C. glutamicum F (designated strain FVan) cultured at 30 °C in AF medium containing FA. Strain C. glutamicum FVan converted 4.57 ± 0.07 mM of FA into 2.87 ± 0.01 mM PCA after 48 h with yields of 62.8% (mol/mol), and 6.91 mM (1064 mg/L) of PCA was produced from 16.0 mM of FA after 12 h of fed-batch biotransformation. Genomic analysis of C. glutamicum ATCC 21420 revealed that the PCA-utilization genes (pca cluster) were conserved in strain ATCC 21420 and that mutations were present in the PCA importer gene pcaK.

Selective Intermolecular Oxidative Cross-Coupling of Enolates.

Selective intermolecular oxidative cross-coupling of enolates, which is a bond-forming reaction between carbanion equivalents, remains as an unsolved issue despite its potential utility for the direct synthesis of unsymmetrical 1,4-diones. The main difficulty derives from the unavoidable homo-coupling. Our strategy depends on the selective one-electron oxidation of one enolate to afford an electrophilic carbonyl α-radical species, followed by trapping with another enolate. The present study demonstrates the selective oxovanadium(V)-induced cross-coupling between boron and silyl enolates.

Production of P-aminobenzoic acid by metabolically engineered escherichia coli.

The production of chemical compounds from renewable resources is an important issue in building a sustainable society. In this study, Escherichia coli was metabolically engineered by introducing T7lac promoter-controlled aroF(fbr), pabA, pabB, and pabC genes into the chromosome to overproduce para-aminobenzoic acid (PABA) from glucose. Elevating the copy number of chromosomal PT7lac-pabA-pabB distinctly increased the PABA titer, indicating that elevation of 4-amino-4-deoxychorismic acid synthesis is a significant factor in PABA production. The introduction of a counterpart derived from Corynebacterium efficiens, pabAB (ce), encoding a fused PabA and PabB protein, resulted in a considerable increase in the PABA titer. The introduction of more than two copies of PT7lac-pabAB (ce-mod), a codon-optimized pabAB (ce), into the chromosome of a strain that simultaneously overexpressed aroF(fbr) and pabC resulted in 5.1 mM PABA from 55.6 mM glucose (yield 9.2%). The generated strain produced 35 mM (4.8 g L(-1)) PABA from 167 mM glucose (yield 21.0%) in fed-batch culture.

Oxovanadium(V)-induced diastereoselective oxidative homocoupling of boron enolates.

Oxovanadium(v)-induced dl-selective oxidative coupling of (Z)-boron enolate was demonstrated to give the corresponding 2,3-disubstituted 1,4-diketone in a good yield. High selectivity (up to 94 : 6) was attained when the reaction was performed with VO(OPr-i)2Cl at -30 °C.

Quantification of the secondary flow in a radial coupled centrifugal blood pump based on particle tracking velocimetry.

Secondary flow in the centrifugal blood pump helps to enhance the washout effect and to minimize thrombus formation. On the other hand, it has an adverse effect on pump efficiency. Excessive secondary flow may induce hemolytic effects. Understanding the secondary flow is thus important to the design of a compact, efficient, biocompatible blood pump. This study examined the secondary flow in a radial coupled centrifugal blood pump based on a simple particle tracking velocimetry (PTV) technique. A radial magnetically coupled centrifugal blood pump has a bell-shaped narrow clearance between the impeller inner radius and the pump casing. In order to vary the flow levels through the clearance area, clearance widths of 0.25 mm and 0.50 mm and impeller washout holes with diameters of 0 mm, 2.5 mm, and 4 mm were prepared. A high-speed video camera (2000 frames per second) was used to capture the particle images from which radial flow components were derived. The flow in the space behind the impeller was assumed to be laminar and Couette type. The larger the inner clearance or diameter of washout hole, the greater was the secondary flow rate. Without washout holes, the flow behind the impeller resulted in convection. The radial flow through the washout holes of the impeller was conserved in the radial as well as in the axial direction behind the impeller. The increase in the secondary flow reduced the net pump efficiency. Simple PTV was successful in quantifying the flow in the space behind the impeller. The results verified the hypothesis that the flow behind the impeller was theoretically Couette along the circumferential direction. The convection flow observed behind the impeller agreed with the reports of other researchers. Simple PTV was effective in understanding the fluid dynamics to help improve the compact, efficient, and biocompatible centrifugal blood pump for safe clinical applications.