Pullulan degrading enzymes belong to a group of glycosylhydrolases that are widely distributed in nature and are produced by an extremely wide variety of species. Among them the thermophilic and mesophilic bacteria are a rich source of these enzymes. There are many biotechnological applications for these enzymes and a rapidly growing amount of information about their diversity, genetic as well as biochemical and biophysical characteristics. The properties of these enzymes vary and are somewhat linked to the natural environment inhabited by the producing organisms. Genes for these enzymes have been cloned from several strains and their amino acid sequences show highly conserved regions common to the enzymes of the amylase family. Molecular studies have greatly extended our knowledge on pullulan degrading enzymes and their biosynthesis. However, enzyme production levels have usually not been as high as had been assumed possible, and the properties of some enzymes are less than optimal for their industrial applications. Some of these problems can be overcome with the use of good producer organisms, optimized expression/secretion vectors, and site-directed mutagenesis. The molecular biology of pullulan degrading enzymes has been and continues to be a valuable system for studying basic questions of cell biology, such as mechanisms of gene regulation and secretion, and the structure–function relationships of proteins.

Probiotics improve consumers' health and additionally may positively influence the microbiological and organoleptic quality of food. In the study, legume sprouts were inoculated with Lactobacilllus plantarum 299V to produce a new functional product ensuring the growth and survival of the probiotic and high microbiological quality of the final product. Legume sprouts, which are an excellent source of nutrients, were proposed as alternative carriers for the probiotic. The key factors influencing the production of probiotic-rich sprouts include the temperature (25°C) of sprouting and methods of inoculation (soaking seeds in a suspension of probiotics). Compared to the control sprouts, the sprouts enriched with the probiotic were characterized by lower mesophilic bacterial counts. In the case of fresh and stored probiotic-rich sprouts, lactic acid bacteria (LAB) accounted for a majority of total microorganisms. The Lb. plantarum population was also stable during the cold storage. The high count of LAB observed in the digest confirmed the fact that the studied sprouts are effective carriers for probiotics and ensure their survival in the harmful conditions of the digestive tract in an in vitro model. Enrichment of legume sprouts with probiotics is a successful attempt and yields products for a new branch of functional foods.

The genus Lactobacillus includes, among others, Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus, species that are collectively referred to as the Lactobacillus casei group. Many studies have shown that strains belonging to this group may decrease lactose intolerance, the effects of inflammatory bowel disease, diarrhea, constipation, food allergies and even colon cancer. Moreover, evidences exists of positive effects of these bacteria on mucosal immunity and blood cholesterol level. Because of their beneficial influence on human health, many of them are used as food additives and probiotic pharmaceuticals. It should be stressed that health-promoting properties are not attributed at the species level, but to specific strains. Therefore, procedures are necessary to allow specific identification at each phylogenetic level—genus, species and strain. In this paper we present a practical overview of molecular methods for the identification and differentiation of L. casei bacteria. The research included 30 bacterial strains belonging to three species: L.casei, L. paracasei and L. rhamnosus. Among the tested procedures were genus- and species-specific PCR, multiplex-PCR, Real-Time HRM analysis, RFLP-PCR, rep-PCR, RAPD-PCR, AFLP-PCR, and proteomic methods such as MALDI-TOF MS typing and SDS-PAGE fingerprinting. The obtained results showed that multiplex-PCR and MALDI-TOF MS turned out to be the most useful methods to identify the tested bacteria at the species level. At the strain level, the AFLP-PCR method showed the highest discriminatory power. We hope that the presented results will allow for the easy selection of an appropriate procedure, depending on the experiment conducted and the equipment capabilities of any given laboratory.

Lactococcus lactis bacteria are used as starter cultures in cottage cheese and fermented milk production due to their acidification and contribution in the creation of the characteristic sensory features. The aim of the research was to carry out isolation, genetic identification, and verification of the probiotic properties of selected Lactococcus lactis isolates obtained from Polish fermented cow milk and apply the best strains to produce cottage cheese with good rheological and sensory characteristics. The isolates obtained were identified morphologically, biochemically, and with the use of 16S rRNA gene sequencing. After pre-screening two of the tested Lactococcus lactis strains, A13 and A14 were observed to be most tolerant to high NaCl concentrations and bile salts and to acidify milk the most. We confirmed the activity of A13 and A14 against such pathogenic strains as Escherichia coli, Salmonella enterica ssp. enterica, Salmonella enterica ssp. enterica sv. anatum, Staphylococcus aureus, and Enterococcus faecalis. As a potential industrial starter culture with probiotic potential, the selected Lactococcus lactis A13 and A14 strains produced cottage cheese quickly with good sensory (colour, smell, taste, texture) and rheological (viscosity, elasticity) properties.