Meat fermentations with immobilized lactic acid bacteria

Summary Two meat starter cultures, one ofLactobacillus plantarum and the otherPediococcus pentosaceus, were immobilized in calcium alginate beads and then lyophilized. Upon inoculation into meat, the immobilized cultures were found to ferment more rapidly than comparable free cell cultures.     

Comparison of isotopic fractionation in lactic acid and ethanol fermentations

Pure D(-) and L(+) enantiomers of lactic acid were prepared by fermentation reactions with specific bacteria. In addition, naturally deuterated ethanol was prepared and converted into diastereoisomers using mandelic acid. Various sugars and nutrients were fermented into lactic acid in water having different deuterium contents and ethanol samples were obtained from yeast fermentation of sugars from different botanical origins. The methine and methylene groups in lactic acid and ethanol respectively show similar deuterium contents which are related to that found in the fermentation water. However, the methyl groups of both molecules are significantly different whatever the botanical origin of the carbon source in the fermentation medium.     

On-line estimation of biological variables during pH controlled lactic acid fermentations

Indirect measurement of lactose, galactose, lactic acid, and biomass concentration from on-line sodium hydroxide weight measurements have been obtained for pure and mixed batch cultures of Streptococcus salivarius ssp. thermophilus 404 and Lactobacillus delbrueckii subsp. bulgaricus 398 conducted at controlled pH and temperature. Linear correlations were established between the equivalent sodium hydroxide concentration and the lactose (substrate), galactose and lactic acid (products) concentrations while nonlinear relationships were developed between biomass and lactic acid concentrations. These nonlinear relationships took into account the inhibitory effect of lactic acid on growth and acidification. The indirect measurements of biomass concentration were introduced into a nonlinear estimator of the state variables and of the specific growth and lactic acid production rates. Good agreement was found between estimated and measured biomass concentrations (error index ranging from 10.8% to 12.6%). The results showed the feasibility of on-line estimation of biomass concentration and of the specific kinetics from NaOH addition weight measurements and its applicability for monitoring lactic acid fermentations. Using off-line measurements of L(+) and D(-) lactic acid concentrations, the evolution of the concentration of each strain in mixed cultures was obtained from the relationships proposed for the mixed cultures. (c) 1994 John Wiley & Sons, Inc.     

Antagonistic activities of lactic acid bacteria in food and feed fermentations

Abstract Many factors contribute to a successful natural fermentation of carbohydrate-rich food and feed products. Metabolic activities of lactic acid bacteria (LAB) play a leading role. Their ability to rapidly produce copious amounts of acidic end products with a concomitant pH reduction is the major factor in these fermentations. Although their specific effects are difficult to quantitate, other LAB metabolic products such as hydrogen peroxide and diacetyl can also contribute to the overall antibiosis and preservative potential of these products. The contribution of bacteriocins is also difficult to evaluate. It is suggested that they may play a role in selecting the microflora which initiates the fermentation. Bacteriocins are believed to be important in the ability of LAB to compete in non-fermentative ecosystems such as the gastro-intestinal tract. During the past few decades interest has arisen in the use of the varied antagonistic activities of LAB to extent the shelf-life of protein-rich products such as meats and fish. Recent findings indicate that the newly discovered Lactobacillus reuteri reuterin system may be used for this purpose.     

Bacteriophages of lactic acid bacteria and their impact on milk fermentations

Every biotechnology process that relies on the use of bacteria to make a product or to overproduce a molecule may, at some time, struggle with the presence of virulent phages. For example, phages are the primary cause of fermentation failure in the milk transformation industry. This review focuses on the recent scientific advances in the field of lactic acid bacteria phage research. Three specific topics, namely, the sources of contamination, the detection methods and the control procedures will be discussed.     

Antagonistic activities of lactic acid bacteria in food and feed fermentations

Many factors contribute to a successful natural fermentation of carbohydrate-rich food and feed products. Metabolic activities of lactic acid bacteria (LAB) play a leading role. Their ability to rapidly produce copious amounts of acidic end products with a concomitant pH reduction is the major factor in these fermentations. Although their specific effects are difficult to quantitate, other LAB metabolic products such as hydrogen peroxide and diacetyl can also contribute to the overall antibiosis and preservative potential of these products. The contribution of bacteriocins is also difficult to evaluate. It is suggested that they may play a role in selecting the microflora which initiates the fermentation. Bacteriocins are believed to be important in the ability of LAB to compete in non-fermentative ecosystems such as the gastrointestinal tract. During the past few decades interest has arisen in the use of the varied antagonistic activities of LAB to extend the shelf-life of protein-rich products such as meats and fish. Recent findings indicate that the newly discovered Lactobacillus reuteri reuterin system may be used for this purpose.     

Physiology of spermatozoa at high dilution rates: the influence of seminal plasma

Extensive dilution of spermatozoa, as occurs during flow-cytometric sperm sorting, can reduce their motility and viability. These effects may be minimized by the use of appropriate dilution and collection media, containing balanced salts, energy sources, egg yolk and some protein. Dilution and flow-cytometric sorting of spermatozoa, which involves the removal of seminal plasma, also destabilizes sperm membranes leading to functional capacitation. This membrane destabilization renders the spermatozoa immediately capable of fertilization in vitro, or in vivo after deposition close to the site of fertilization, but shortens their lifespan, resulting in premature death if the cells are deposited in the female tract distant from the site of fertilization or are held in vitro at standard storage temperatures. This functional capacitation can be reversed in boar spermatozoa by inclusion of seminal plasma in the medium used to collect the cells from the cell sorter and, consequently, reduces their in vitro fertility. It has yet to be determined whether seminal plasma would have similar effects on flow cytometrically sorted spermatozoa of other species, and what its effects might be on the in vivo fertility of flow sorted boar.     

Genome analysis of lactic acid bacteria in food fermentations and biotechnological applications

Lactic acid bacteria are an important group of microorganisms, several of which are used in fermented food processes. Lactococcus lactis is a non-pathogenic, non-invasive and non-colonising gram-positive lactic acid bacterium, the genome sequence of which has been established. A great deal is known about the genetics, vectors, gene expression systems and protein secretion apparatus of this bacterium. Recently, recombinant strains of L. lactis have been developed that might provide in vivo delivery of cytokines and specific antigens across mucosal surfaces to the immune system of animals.     

Influence of dilution rate and induction of cloned gene expression in continuous fermentations of recombinant yeast

The effects of growth rate on cloned gene product synthesis in recombinant Saccharomyces cerevisiae have been studied in continuous culture. The plasmid employed contains a yeast GAL10-CYC1 hybrid promoter directing expression of the E. coli lacZ gene. beta-Galactosidase production was therefore controlled by the yeast galactose regulatory circuit, and the induction process and its effects were studied at the various dilution rates. At all dilution rates plasmid stability decreased with induction of lacZ gene expression. In some instances, two induced "steady states" were observed, the first 10-15 residence times after induction and the second after 40-50 residence times. The second induced steady state was characterized by greater biomass concentration and lower beta-galactosidase specific activity relative to the first induced "steady-state." beta-Galactosidase specific activity and biomass concentration increased as dilution rate was reduced, and despite lower flow rate and plasmid stability, overall productivity (activity/L/hr) was substantially higher at low dilution rate. Important factors influencing all of the trends were the glucose and galactose (inducer) concentrations in the vessel and inducer metabolism.     

Evaluation of technologically important traits in lactic acid bacteria isolated from spontaneous fermentations

A set of 120 isolates of Lactobacillus (75), Lactococcus (25) and Leuconostoc (20), mainly isolated from natural starter-free fermentation processes, has been screened for some technologically important traits, including lactose hydrolysis, extracellular polysaccharide production, fermentation of carbohydrates, antimicrobial agent susceptibility and bacteriocin production. Some of the strains exhibited important features for particular fermentations. Furthermore, the sensitivity resistance phenotype to antimicrobials showed genus-specific patterns.     

The effect of equipment scale and degree of mixing on continuous fermentation yield at low dilution rates

Previously, the degree of mixing was not felt to be an important consideration in fermentor design. In this study on the continuous propagation of Baker's yeast, it was found that at low dilution rates, i.e., 0.02hr^?1, the degree of mixing achieved does effect the cell yield. At low dilution rates, appreciable quantities of sugar can be utilized for endogenous respiration in comparison to that utilized for making cell mass. Poor distribution of the sugar aggravates the balance of sugar utilized for each process. Yields at these low dilution rates can be improved to a limited extent by using a multiple feed-distribution system and better mixing.     

Simultaneous and sequential fermentations with yeast and lactic acid bacteria in apple juice

A complex substrate, reconstituted concentrated apple juice, was used for testing the principal processes during yeast and malolactic bacteria fermentations. Interactions between microorganisms were studied based on two controlled inoculation procedures, and at different fermentation temperatures. Temperature had a more important effect on yeast growth than the presence of malolactic bacteria in the medium. Acceleration of the death phase of the bacterial population was detected at increased temperatures. In all cases, malic acid degradation was affected by the fermentation temperature. When experiments were carried out with simultaneous inoculation, acidification of the medium took place at both temperatures tested (15°C and 22°C), that was not observed when the malolactic bacteria were inoculated after completion of alcoholic fermentation by yeasts. Received 4 August 1998/ Accepted in revised form 9 December 1998     

Production of acetic acid by Clostridium thermoaceticum in batch and continuous fermentations

Batch and continuous fermentations with Clostridium thermoaceticum (ATCC 39073) using automatic pH control were conducted. The value of mu(max) obtained from batch fermentation was about 0.14 h(-1); acetate yield, which was both growth and non-growth associated, was about 2 mole of acetic acid/mole of glucose, compared with a theoretical maximum value of 3. This low yield, compared with literature data, may be explained by glucose loss through a combination of degradation routes. Continuous fermentation could be sustained for 1600 h or more without contamination problems. Continuous fermentation at high dilution rates indicates that mu(max) may be well above 0.17 h(-1) when fresh feed medium is used. Acetate yields in continuous fermentation were about 77% of theoretical or 2.3 mole of acetic acid/mole of glucose.     

Evaluation of productive biofilms for continuous lactic acid production

In white biotechnology research, the putative superiority of productive biofilms to conventional biotransformation processes based on planktonic cultures has been increasingly discussed in recent years. In the present study, we chose lactic acid production as a model application to evaluate biofilm potential. A pure culture of Lactobacillus bacteria was grown in a tubular biofilm reactor. The biofilm system was cultivated monoseptically in a continuous mode for more than 3 weeks. The higher cell densities that could be obtained in the continuous biofilm system compared with the planktonic culture led to a significantly increased space-time yield. The productivity reached 80% of the maximum value 10 days after start-up and no subsequent decline was observed, confirming the suitability of the system for long-term fermentation. The analysis of biofilm performance revealed that productivity increases with the flow velocity. This is explained by the reduced retention time of the liquid phase in the reactor, and, thus, a minor pH drop caused by the released lactic acid. At low flow velocities, the pH drops to a value where growth and production are significantly inhibited. The biofilm was visualized by magnetic resonance imaging to analyze biofilm thickness. To deepen the understanding of the biofilm system, we used a simple model for cell growth and lactic acid production.     

Batch fermentations on synthetic mixed sugar and starch medium with amylolytic lactic acid bacteria

The green crop drying industry in Denmark uses Italian rye grass, clover, and alfalfa as raw materials for the production of green pellets. The green crop drying industry solves its energy economical problems by heating and pressing of the green crop before drying. The produced sidestream is called brown juice. Brown juice was shown to be an excellent medium for lactic acid fermentation. The aim of this study was to investigate the utilisation of brown juice in the production of polylactic acid, where wheat starch would be added to increase the lactic acid yield and, thus, the feasibility of the process. A number of amylolytic lactic acid bacteria have been identified, and in this work, six different strains were tested for their ability to produce α-amylase and to utilise all sugars with high lactic acid yield in a medium with a complex composition of free sugars (brown juice) and starch. Lactobacillus plantarum A6 was the only strain that showed both a good lactic acid production and utilisation of starch in this medium. The growth rate of this strain was approximately 0.4 h⁻¹ and the lactic acid yield was 0.7.     

Stable production of hyaluronic acid in Streptococcus zooepidemicus chemostats operated at high dilution rate

Hyaluronic acid is routinely produced through fermentation of both Group A and C streptococci. Despite significant production costs associated with short fermentations and removal of contaminating proteins released during entry into stationary phase, hyaluronic acid is typically produced in batch rather than continuous culture. The main reason is that hyaluronic acid synthesis has been found to be unstable in continuous culture except at very low dilution rates. Here, we investigated the mechanisms underlying this instability and developed a stable, high dilution rate (0.4 h-1) chemostat process for both chemically defined and complex media operating for more than 150 h of production. In chemically defined medium, the product yield was 25% higher in chemostat cultures than in conventional batch culture when arginine or glucose was the limiting substrate. In contrast, glutamine limitation resulted in higher ATP requirements and a yield similar to that observed in batch culture. In complex, glucose-limited medium, ATP requirements were greatly reduced but biomass synthesis was favored over hyaluronic acid and no improvement in hyaluronic acid yield was observed. The successful establishment of continuous culture at high dilution rate enables both commercial production at reduced cost and a more rational characterization and optimization of hyaluronic acid production in streptococci.     

Genetic aspects of aromatic amino acid biosynthesis in Lactococcus lactic

Polymerase chain reaction (PCR) primers designed from a multiple alignment of predicted amino acid sequences from bacterial aroA genes were used to amplify a fragment of Lactococcus lactis DNA. An 8 kb fragment was then cloned from a lambda library and the DNA sequence of a 4.4 kb region determined. This region was found to contain the genes tyrA, aroA, aroK, and pheA, which are involved in aromatic amino acid biosynthesis and folate metabolism. TyrA has been shown to be secreted and AroK also has a signal sequence, suggesting that these proteins have a secondary function, possibly in the transport of amino acids. The aroA gene from L. lactis has been shown to complement an E. coli mutant strain deficient in this gene. The arrangement of genes involved in aromatic amino acid biosynthesis in L. lactis appears to differ from that in other organisms.