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Malligarjunan Rajavel, Kumar Perinbam, B Gopal
The synthesis of the dipeptide antibiotic bacilysin involves the sequential action of multiple enzymes in the bac operon. YwfH (also referred to as BacG) catalyzes the stereoselective reduction of dihydro-hydroxyphenylpyruvate (H2HPP) to tetrahydro-hydroxyphenylpyruvate (H4HPP) in this biosynthetic pathway. YwfH is an NADPH-dependent reductase that facilitates the conjugate addition of a hydride at the C4 olefin terminus of H2HPP. Here, the structure of YwfH is described at three conformational steps: the apo form, an apo-like conformation and the NADPH complex...
March 2013: Acta Crystallographica. Section D, Biological Crystallography
Jared B Parker, Christopher T Walsh
The first four enzymes of the bacilysin antibiotic pathway, BacABGF, convert prephenate to a tetrahydrotyrosine (H(4)Tyr) diastereomer on the way to the anticapsin warhead of the dipeptide antibiotic. BacB takes the BacA product endocyclic-Δ(4),Δ(8)-7R-dihydrohydroxyphenylpyruvate (en-H(2)HPP) and generates a mixture of 3E- and 3Z-olefins of the exocyclic-Δ(3),Δ(5)-dihydrohydroxyphenylpyruvate (ex-H(2)HPP). The NADH-utilizing BacG then catalyzes a conjugate reduction, adding a pro-S hydride equivalent to C(4) to yield tetrahydrohydroxyphenylpyruvate (H(4)HPP), a transamination away (via BacF) from 2S-H(4)Tyr...
July 17, 2012: Biochemistry
P D Constable, W Grünberg, L Carstensen
The effect of adding an oral rehydration therapy (ORT) solution containing bicarbonate, citrate, and acetate to cow's milk on milk clotting, abomasal luminal pH, and abomasal emptying rate was determined in suckling calves. Six male Holstein-Friesian calves with abomasal cannulae, 5 to 13 d of age, were used in a crossover design. Calves were fed 2 L of cow's milk alone, milk with an ORT solution containing a low bicarbonate concentration (25 mmol/L), acetate (12 mmol/L), citrate (12 mmol/L), and glycine (7 mmol/L; group BACG), or milk with an ORT solution containing formate (58 mmol/L) and acetate (15 mmol/L) in randomized order...
January 2009: Journal of Dairy Science
S W Park, H K Lee, T G Kim, S K Yoon, S Y Paik
We have developed the recombinant baculovirus pseudotyped with vesicular stomatitis virus (VSV) G protein. The VSV-G gene was under the control of the polyhedrin promoter so that it was expressed at high levels in infected insect cells but not in mammalian cells. The presence of VSV-G protein in purified baculovirus preparations was confirmed by Western analysis. This recombinant baculovirus also carried human AFP (alpha-fetoprotein) promoter for hepatocyte-specific gene expression. After an in vitro infection by a recombinant baculovirus carrying the luciferase gene under the control of human AFP promoter/enhancer (BacG-AFP-Luc(+)), the luciferase gene was expressed in AFP-producing Huh7, Hep3B, and HepG2 cell lines, but not in AFP-nonproducing cell lines...
November 30, 2001: Biochemical and Biophysical Research Communications
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