Genes for the biosynthesis of spinosyns: applications for yield improvement in Saccharopolyspora spinosa

Spinosyns A and D are the active ingredients in an insect control agent produced by fermentation of Saccharopolyspora spinosa. Spinosyns are macrolides with a 21-carbon, tetracyclic lactone backbone to which the deoxysugars forosamine and tri-O-methylrhamnose are attached. The spinosyn biosynthesis genes, except for the rhamnose genes, are located in a cluster that spans 74 kb of the S. spinosa genome. DNA sequence analysis, targeted gene disruptions and bioconversion studies identified five large genes encoding type I polyketide synthase subunits, and 14 genes involved in sugar biosynthesis, sugar attachment to the polyketide or cross-bridging of the polyketide. Four rhamnose biosynthetic genes, two of which are also necessary for forosamine biosynthesis, are located outside the spinosyn gene cluster. Duplication of the spinosyn genes linked to the polyketide synthase genes stimulated the final step in the biosynthesis — the conversion of the forosamine-less pseudoaglycones to endproducts. Duplication of genes involved in the early steps of deoxysugar biosynthesis increased spinosyn yield significantly. Journal of Industrial Microbiology & Biotechnology (2001) 27, 399–402. Received 31 May 2001/ Accepted in revised form 09 July 2001     

Molecular cloning and DNA sequence analysis of the human guanine nucleotide-binding protein Go alpha

The nucleotide sequence of human Go alpha was determined from a partial human brain cDNA clone and the sequence of the first two 5' coding exons of a human genomic Go alpha clone. Comparison of this sequence with bovine and rat Go alpha shows greater than 90% homology at the nucleotide and deduced amino acid level. There is 100% identity at the amino acid level for the cholera and pertussis toxin-catalyzed ADP ribosylation sites, the putative guanine nucleotide binding, and the GTP hydrolysis sites.     

A comment on the evaluation of equilibrium constants for alpha-tocopherol interactions with fatty acids by absorbance in the ultraviolet region

This paper comments on the evaluation of Erin and co-workers (Biochim. Biophys. Acta 774 (1984) 96-102) of equilibrium constants for alpha-tocopherol interactions with fatty acids on the basis of the changes of absorbance in a 200 nm ultraviolet region. It is concluded that the ultraviolet method is inadequate because it is affected by absorption in that region of the solvent, ethanol and fatty acids which they used.     

An attempt to determine lipid peroxides with polyethylene glycol-modified hemin

Hemin, having two carboxyl groups, was coupled with alpha-(3-aminopropyl)-omega-methoxypoly(oxyethylene) through the acid-amide bond formed with carbodiimide. The modified hemin catalyzed the peroxidase reaction in 1,1,1-trichloroethane using benzoyl peroxide or peroxides in unsaturated fatty acids as the hydrogen acceptor and leuco crystal violet as the hydrogen donor. A basic study on quantitative microanalysis of the lipid peroxides was attempted.