Biogenic amines as an indicator of rye leaven quality during production and storage.

Rye leaven, the basic constituent of sour rye soup ('zurek' or white borsch), was obtained through three methods of initiating lacto-fermentation of rye flour. Optimal concentrations of NaCl (1.5%) and garlic (0.5%) were selected by utilizing the response surface methodology. During the production and storage of leaven at 10 °C and 20 °C, the secalin proteins of rye flour degraded significantly and the concentration of free amino acids increased, making the rye leaven an environment potentially conducive to the formation of biogenic amines. Putrescine (max. conc: 116.7 mg kg-1) and tyramine (max. conc: 63.4 mg kg-1) were the amines that occurred in the largest amounts in the leavens. The final concentration of histamine (after 150 days of storage) did not exceed 22 mg kg-1. Regardless of the method of initiation of fermentation, the products that contained fewer biogenic amines better retained their sensory characteristics (r ≤ -0.89, p < 0.05) and had a higher number of lactic acid bacteria (r ≤ -0.66, p < 0.05).

Incorporation of reporter genes into mitochondrial DNA in budding yeast.

Many aspects of mitochondrial gene expression are still unknown, which can be attributed to limitations in molecular tools. Here, we present a protocol to introduce reporter genes into the mitochondrial genome of budding yeast, Saccharomyces cerevisiae. Mitochondrially encoded reporter constructs can be used to interrogate various aspects of mitochondrial gene expression. The power of this technique is exemplified by a mitochondrially encoded nanoluciferase, which allows to monitor levels of mitochondrial translation under a variety of growth conditions.

Nucleotide degradation, biogenic amine level and microbial contamination as quality indicators of cold-stored rainbow trout (Oncorhynchus mykiss) gravad.

The extent to which the transformation of nucleotides, biogenic amines, and microbiological changes affect the quality and shelf life of vacuum packaged low processed rainbow trout (Oncorhynchus mykiss) gravad during storage at 7 ± 1 °C for 42 days was investigated. Although total viable counts increased slowly up to 6 log CFU g-1 at the end of storage, coliform bacteria disappeared. The histamine concentration and the biogenic amine index increased up to 45.2 ± 1.62 mg kg-1 and 100 mg kg-1 respectively. The highest concentration of inosine monophosphate was achieved in freshly prepared gravad, whereas the hypoxanthine level increased with storage time up to 28 days. Among nucleotide ratios, the G value is more suitable for the determination of gravad quality than K, Ki and H values. Once the gravad obtained the limit of acceptability by the panelists (35 days) the G value rose to 470%.

The Analysis of Yeast Mitochondrial Translation.

The mitochondrial genome encodes only a handful of proteins, but methods to track their synthesis are highly limited. Saccharomyces cerevisiae is a model organism that offers possibilities to expand the classical systems to analyze mitochondrial translation. In this chapter, we present two approaches of monitoring mitochondrial protein synthesis. Labeling of mitochondrially translated products with radioactive amino acids can be performed either in intact cells or in isolated mitochondria. However, these classical methods have disadvantages that can affect cell physiology and hence are not suitable for all types of research questions. Some of these limitations can be overcome by the use of reporter genes that are inserted into yeast genetic screens mitochondrial DNA via biolistic transformation. These reporter genes can be used for yeast genetic screen and to monitor regulation and efficiency of mitochondrial translation with a variety of methods.

A novel system to monitor mitochondrial translation in yeast.

The mitochondrial genome is responsible for the production of a handful of polypeptides that are core subunits of the membrane-bound oxidative phosphorylation system. Until now the mechanistic studies of mitochondrial protein synthesis inside cells have been conducted with inhibition of cytoplasmic protein synthesis to reduce the background of nuclear gene expression with the undesired consequence of major disturbances of cellular signaling cascades. Here we have generated a system that allows direct monitoring of mitochondrial translation in unperturbed cells. A recoded gene for superfolder GFP was inserted into the yeast (Saccharomyces cerevisiae) mitochondrial genome and enabled the detection of translation through fluorescence microscopy and flow cytometry in functional mitochondria. This novel tool allows the investigation of the function and regulation of mitochondrial translation during stress signaling, aging and mitochondrial biogenesis.