ABSTRACT
Stability and radical scavenging activity of betalains, biosynthesized by Beta vulgaris hairy root culture were investigated at different pH values, bile salts concentration, as well as at the in vitro conditions of gastrointestinal tract. It was established that pH below 3 and concentrations of the bile salts up to 4% had no great influence on betalains stability. At the in vitro conditions of gastrointestinal tract betalains are relatively stable, as their radical scavenging activity decrease from 75% inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH*) to about 38%.
Subject(s)
Beta vulgaris/chemistry , Free Radical Scavengers/metabolism , Gastrointestinal Contents/chemistry , Quaternary Ammonium Compounds/metabolism , Betalains , Cells, Cultured , Humans , Hydrogen-Ion Concentration , Models, Biological , Oxidation-Reduction , Plant Roots/chemistryABSTRACT
A mutant of the methylotrophic yeast Hansenula polymorpha with constitutive alcohol oxidase (AOX) and peroxisome biosynthesis was obtained after UV treatment followed by cell plating on a medium containing methanol and 2-deoxy-D-glucose (DOG). DOG-resistant colonies of mutants were insensitive to catabolic repression by glucose and methanol. A selection procedure is described that allows the isolation of a mutant exhibiting a constitutive phenotype of AOX involved in methanol utilization. Furthermore, additional features of the constitutive presence of peroxisomes are demonstrated. 562 DOG-resistant colonies were tested, 24 of them demonstrating constitutive AOX formation. Based on quantitative analysis, one of the strains--DOG-13 was selected and its growth, biochemical and ultrastructural characteristics were examined. Its specific enzyme activity when cultivated on a yeast nitrogen base + 1% glucose (YNB + 1% Glucose) was found to reach 145 nmol x min(-1) x mg(-1) protein (compared to zero of the parent strain) after he 20th hour of cultivation. This was confirmed by fine-structure analysis, showing typical peroxisomes, which number and size increased with the enzyme activity. This study demonstrates a constitutive AOX and peroxisome biosynthesis by the mutant strain H. polymorpha DOG-13 obtained.
