D. van Sinderen

B-group vitamin production by lactic acid bacteria--current knowledge and potential applications.

Although most vitamins are present in a variety of foods, human vitamin deficiencies still occur in many countries, mainly because of malnutrition not only as a result of insufficient food intake but also because of unbalanced diets. Even though most lactic acid bacteria (LAB) are auxotrophic for several vitamins, it is now known that certain strains have the capability to synthesize water‐soluble vitamins such as those included in the B‐group (folates, riboflavin and vitamin B12 amongst others). This review article will show the current knowledge of vitamin biosynthesis by LAB and show how the proper selection of starter cultures and probiotic strains could be useful in preventing clinical and subclinical vitamin deficiencies. Here, several examples will be presented where vitamin‐producing LAB led to the elaboration of novel fermented foods with increased and bioavailable vitamins. In addition, the use of genetic engineering strategies to increase vitamin production or to create novel vitamin‐producing strains will also be discussed. This review will show that the use of vitamin‐producing LAB could be a cost‐effective alternative to current vitamin fortification programmes and be useful in the elaboration of novel vitamin‐enriched products.

Lactic acid bacteria with potential to eliminate fungal spoilage in foods

Aims:  To investigate antifungal activity produced by lactic acid bacteria (LAB) isolated from malted cereals and to determine if such LAB have the capacity to prevent fungal growth in a particular food model system.

Characterization of bacteriocin-like inhibitory substances (BLIS) from sourdough lactic acid bacteria and evaluation of their in vitro and in situ activity

Aims:  To identify and characterize bacteriocion‐producing lactic acid bacteria (LAB) in sourdoughs and to compare in vitro and in situ bacteriocin activity of sourdough‐ and nonsourdough LAB.

An analysis of bacteriocins produced by lactic acid bacteria isolated from malted barley

Aims: The aim of this study was to perform a detailed characterization of bacteriocins produced by lactic acid bacteria (LAB) isolated from malted barley.

Selective carbohydrate utilization by lactobacilli and bifidobacteria

To evaluate the ability of specific carbohydrates, including commercially available products, to support the growth of representatives of two well‐known groups of gut commensals, namely lactobacilli and bifidobacteria.

Six putative two-component regulatory systems isolated from Lactococcus lactis subsp. cremoris MG1363

The genetic elements specifying six putative two-component regulatory systems (2CSs) were identified on the chromosome of Lactococcus lactis MG1363. These 2CSs appear to represent distinct loci, each containing a histidine kinase and response-regulator-encoding gene pair. Transcriptional analysis of the six 2CSs was performed either by generating transcriptional fusions to a reporter gene or by primer extension. Two of the systems appeared to be expressed constitutively at a high level, whilst the remaining four exhibited growth-phase-dependent expression. Insertional mutagenesis established that the two constitutively expressed 2CSs are necessary for normal cell growth and/or survival. Mutational analysis of the remaining four systems revealed that they are implicated in susceptibility to extreme pH, osmotic or oxidative conditions, or the regulation of phosphatase activity in L. lactis.

Molecular genetics of bacteriophage and natural phage defence systems in the genus Lactococcus

Abstract Bacteriophage infection of starter cultures used in a range of milk fermentation processes, particularly those involving Lactococcus lactis, poses a significant problem in industrial practice. The application of genetic and molecular technologies to the study of lactococcal bacteriophages has proven to be very rewarding in terms of understanding the nature of phage with respect to their physical and genetic organisation. The availability of the full genomic sequence of a number of phages provides an unambiguous basis for determining the relationship between them, for elucidating their evolutionary progression and will also yield strategies for obstructing successful phage proliferation on previously sensitive hosts. The genetic analysis of phage/host interactions has also highlighted the presence of natural defence systems (e.g. adsorption blocking, inhibition of phage DNA entry, restriction modification and abortive infection) in lactococci. A number of restriction modification systems and abortive infection mechanisms have been characterized at a molecular level and the genes involved have been cloned and sequenced. Plasmid-encoded phage resistance mechanisms can be exploited to generate strains which can successfully counter phage proliferation and will provide a basis for understanding the complex interactions between phages and their target hosts at a molecular level.

In situ activity of a bacteriocin-producing Lactococcus lactis strain. Influence on the interactions between lactic acid bacteria during sourdough fermentation

Aims:  To biochemically characterize the bacteriocin produced by Lactococcus lactis ssp. lactis M30 and demonstrate its effect on lactic acid bacteria (LAB) during sourdough propagation.

The Streptococcus thermophilus Autolytic Phenotype Results from a Leaky Prophage

ABSTRACT Streptococcus thermophilus autolytic strains are characterized by a typical bell-shaped growth curve when grown under appropriate conditions. The cellular mechanisms involved in the triggering of lysis and the bacteriolytic activities of these strains were investigated in this study. Lactose depletion and organic solvents (ethanol, methanol, and chloroform) were shown to trigger a premature and immediate lysis of M17 exponentially growing cells. These factors and compounds are suspected to act by altering the cell envelope properties, causing either the permeabilization (organic solvents) or the depolarization (lactose depletion) of the cytoplasmic membrane. The autolytic character was shown to be associated with lysogeny. Phage particles, most of which were defective, were observed in the culture supernatants after both mitomycin C-induced and spontaneous lysis. By renaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a bacteriolytic activity was detected at 31 kDa exclusively in the autolytic strains. This enzyme was detected during both growth and spontaneous lysis with the same intensity. We have shown that it was prophage encoded and homologous to the endolysin Lyt51 of the streptococcal temperate bacteriophage φ01205 (M. Sheehan, E. Stanley, G. F. Fitzgerald, and D. van Sinderen, Appl. Environ. Microbiol. 65:569–577, 1999). It appears from our results that the autolytic properties are conferred to the S. thermophilus strains by a leaky prophage but do not result from massive prophage induction. More specifically, we propose that phagic genes are constitutively expressed in almost all the cells at a low and nonlethal level and that lysis is controlled and achieved by the prophage-encoded lysis proteins.

Molecular characterization of the lactococcal plasmid pCIS3: natural stacking of specificity subunits of a type I restriction/modification system in a single lactococcal strain.

A 6.1 kb plasmid from the Lactococcus lactis subsp. cremoris strain UC509.9, named pCIS3, was found to mediate a restriction/modification (R/M) phenotype. Nucleotide sequence analysis of pCIS3 revealed the presence of an hsdS gene, typical of type I R/M systems. The presence of this plasmid resulted in a 10(4)-fold reduction in the efficiency of plating (e.o.p.) of unmodified phage. In addition to the hsdS gene of pCIS3, two more hsdS genes were identified in strain UC509.9, one located on the chromosome downstream of a gene highly homologous to hsdM genes and a third on the smallest (4 kb) plasmid, named pCIS1. The replication region of pCIS3 was highly similar to that of a large family of lactococcal theta replicons. In addition, pCIS3 was found to encode a member of the CorA family of magnesium transporters.