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Taxonomy and important features of probiotic microorganisms in food and nutrition.

Lactic acid bacteria are among the most important probiotic microorganisms typically associated with the human gastrointestinal tract. Traditionally, lactic acid bacteria have been classified on the basis of phenotypic properties, eg, morphology, mode of glucose fermentation, growth at different temperatures, lactic acid configuration, and fermentation of various carbohydrates. Studies based on comparative 16S ribosomal RNA sequencing analysis, however, showed that some taxa generated on the basis of phenotypic features do not correspond with the suggested phylogenetic relations. Thus, some species are not readily distinguishable by phenotypic characteristics. This is especially true for the so-called Lactobacillus acidophilus group, the Lactobacillus casei and Lactobacillus paracasei group, and some bifidobacteria, strains of which have been introduced in many probiotic foods, eg, the novel yogurt-like commodities. Consequently, modern molecular techniques, including polymerase chain reaction-based and other genotyping methods, have become increasingly important for species identification or for the differentiation of probiotic strains. Probiotic strains are selected for potential application on the basis of particular physiologic and functional properties, some of which may be determined in vitro. The classification and identification of a probiotic strain may give a strong indication of its typical habitat and origin. The species, or even genus name, may also indicate the strain's safety and technical applicability for use in probiotic products. Molecular typing methods such as pulsed-field gel electrophoresis, repetitive polymerase chain reaction, and restriction fragment length polymorphism are extremely valuable for specific characterization and detection of such strains selected for application as probiotics.

Bifidobacterium↗

Influence of a probiotic yoghurt on the status of vitamins B(1), B(2) and B(6) in the healthy adult human.

BACKGROUND/AIMS: The main reason for this study was to determine whether yoghurt bacteria, being rich in some water-soluble vitamins, release them or utilize vitamins from their surroundings. Our study was trying to determine for the first time, if the viable bacteria of probiotic yoghurt are able to influence the parameters of the B-vitamin (B(1), B(2), B(6)) status of the healthy adult human. METHODS: The test yoghurt was commercially available probiotic yoghurt prepared with Streptococcus thermophilus and Lactobacillus acidophilus, enriched with Lactobacillus casei GG. Different chemical forms of all investigated B-vitamins were determined by HPLC methods. In order to determine the influence of the yoghurt flora, each of 12 subjects consumed four yoghurt portions 125 g each ( = 500 g) a day, containing thermally inactivated cultures during the first 2-week period and yoghurt without heat treatment during the second 2-week period. RESULTS: The heat treatment of the probiotic yoghurt caused negligible changes in vitamin contents. The plasma levels of thiamin decreased significantly (p < 0.01) after the first 2-week period and kept on decreasing during the second 2-week period. A similar trend was found in the urinary excretion. The plasma levels of the B(2)-vitamers were different. The flavin adenine dinucleotide concentrations increased significantly (p < 0.01) after the consumption of heat-treated yoghurt and decreased significantly (p < 0.05) after the following 2 weeks, in which the subjects received the untreated yoghurt. In contrast, the flavin mononucleotide plasma levels decreased during the first 2-week period and increased during the second part of the study, but the change was not statistically significant. The free riboflavin concentrations in plasma and urine showed a continuous but not significant increase. The concentrations of pyridoxal-5-phosphate in plasma increased after the consumption of yoghurt with the inactivated bacteria and decreased in the second part of the study. However, the differences were not significant. The excretion of thiamin, B(2)- and B(6)-vitamers in the faeces did not significantly change throughout the study period (p>0.05). CONCLUSIONS: Our observations show that the bacterial flora of the examined yoghurt does not influence the vitamin B(1), B(2) and B(6) status of man. It seems likely that even lactobacilli of the 'probiotic' type which are vitamin B consumers can decrease the bioavailability of these vitamins for man. Obviously a thermal death of the cells did not induce a release of physiologically active vitamins.

Adult↗

Characterization of the lactic acid bacteria in ewe's milk and cheese from northwest Argentina.

Indigenous lactic acid bacteria in ewe's milk and artisanal cheese were studied in four samples of fresh raw milk and four 1-month-old cheeses from the provinces of northwest Argentina. Mean growth counts on M17, MRS, and MSE agar media did not show significant differences (P < 0.05) in raw milk and cheeses. Isolates of lactic acid bacteria from milk were identified as Enterococcus (48%), lactococci (14%), leuconostocs (8%), and lactobacilli (30%). All lactococci were identified as Lactococcus lactis (subsp. lactis and subsp. cremoris). Lactobacilli were identified as Lactobacillus plantarum (92%) and Lactobacillus acidophilus (8%). Enterococci (59%) and lactobacilli (41%) were isolated from cheeses. L. plantarum (93%), L. acidophilus (5%), and Lactobacillus casei (2%) were most frequently isolated. L. lactis subsp. lactis biovar diacetylactis strains were considered as fast acid producers. L. lactis subsp. cremoris strains were slow acid producers. L. plantarum and L. casei strains identified from the cheeses showed slow acid production. The majority of the lactobacilli and Lactococcus lactis strains utilized citrate and produced diacetyl and acetoin in milk. Enzyme activities (API-ZYM tests) of lactococci were low, but activities of L. plantarum strains were considerably higher. The predominance of L. plantarum in artisanal cheese is probably important in the ripening of these cheeses due to their physiological and biochemical characteristics.

Animals↗

[Prevention of dysbacteriosis in the early neonatal period using a pure culture of acidophilic bacteria].

In order to prevent dysbacterioses occurring in the early neonatal period, 60 neonates classified with the risk group were administered, when they were still at the puerperal room, pure cultures of specially selected strains of Lactobacillus acidophilus possessing marked anti-Klebsiella activity and antagonistic activity against Staphylococcus aureus. The cultures were instilled into the mouth and nasal passages. Forty neonates administered bifidum-bacterin and 54 neonates not given biopreparations were used as control. Application of the pure culture of Lactobacillus acidophilus noticeably raised the efficacy of the preventive measures. Over 60 percent of the neonates were discharged from the maternity home with a normally formed intestinal microflora, 60 percent had normal microflora in the fauces, 80 percent in the nose, and 70 percent of the neonates demonstrated normal skin microflora. The carriage of opportunistic microorganisms in the fauces and nose of the neonates had decreased substantially by the time of the discharge from the maternity home, while the incidence of pyo-inflammatory diseases had also reduced among them.

Administration, Intranasal↗

Effect of probiotic supplementation on bacterial translocation in thermal injury.

PURPOSE: To examine the effects of probiotic supplementation and enteral solutions containing glutamine and arginine on bacterial translocation (BT) and intestinal villous atrophy in thermal injury. METHODS: Forty male Sprague-Dawley rats weighing 200-250 g were divided into four groups of ten. Group 1 served as control group without thermal injury and was fed standard chow. Thermal injury was inflicted as a 30% scald burn in the other three groups. Group 2 was fed standard chow and group 3 was fed standard chow supplemented with a probiotic (Acidophilus plus) containing Bifidobacterium bifidum, Lactobacillus acidophilus, and Lactobacillus bulgaricus (2 x 10(9) CFU/day) via an orogastric tube. Group 4 was fed only an enteral diet (Stresson multifiber) containing glutamine, arginine, and medium chain triglyceride, at 1 g/kg per day amino acid and 230 kcal/kg, for 7 days before thermal injury. All the animals were killed 24 h after thermal injury, and ileal segments were resected and examined histopathologically. To evaluate BT, samples from blood, mesenteric lymph nodes, and cecal content were cultured under aerobic and anaerobic conditions. Terminal ileum specimens were histologically examined to evaluate mucosal integrity. RESULTS: Significantly less BT was seen in groups 3 and 4 than in group 2 (P < 0.001). No significant difference was found between groups 3 and 4. Histological evaluation showed significant reduction in villous atrophy in groups 3 and 4. CONCLUSION: Probiotic supplementation seems to reduce bacterial translocation and decrease intestinal mucosal atrophy in rats with thermal injury, as do enteral solutions with arginine and glutamine.

Analysis of Variance↗

Cloning, phenotypic expression, and DNA sequence of the gene for lactacin F, an antimicrobial peptide produced by Lactobacillus spp.

Lactacin F is a heat-stable bacteriocin produced by Lactobacillus acidophilus 11088. A 63-mer oligonucleotide probe deduced from the N-terminal lactacin F amino acid sequence was used to clone the putative laf structural gene from plasmid DNA of a lactacin F-producing transconjugant, L. acidophilus T143. One clone, NCK360, harbored a recombinant plasmid, pTRK160, which contained a 2.2-kb EcoRI fragment of the size expected from hybridization experiments. An Escherichia coli-L. acidophilus shuttle vector was constructed, and a subclone (pTRK162) containing the 2.2-kb EcoRI fragment was introduced by electroporation into two lactacin F-negative strains, L. acidophilus 89 and 88-C. Lactobacillus transformants containing pTRK162 expressed lactacin F activity and immunity. Bacteriocin produced by the transformants exhibited an inhibitory spectrum and heat stability identical to those of the wild-type bacteriocin. An 873-bp region of the 2.2-kb fragment was sequenced by using a 20-mer degenerate lactacin F-specific primer to initiate sequencing from within the lactacin F structural gene. Analysis of the resulting sequence identified an open reading frame which could encode a protein of 75 amino acids. The 25 N-terminal amino acids for lactacin F were identified within the open reading frame along with an N-terminal extension, possibly a signal sequence. The lactacin F N-terminal sequence, through the remainder of the open reading frame (57 amino acids; 6.3 kDa), correlated extremely well with composition analyses of purified lactacin F which also predicted a size of 51 to 56 amino acid residues. Molecular characterization of lactacin F identified a small hydrophobic peptide that may be representative of a common bacteriocin class in lactic acid bacteria.

Amino Acid Sequence↗

Mutations within the putative active site of heterodimeric deoxyguanosine kinase block the allosteric activation of the deoxyadenosine kinase subunit.

Replacement of the Asp-84 residue of the deoxyguanosine kinase subunit of the tandem deoxyadenosine kinase/ deoxyguanosine kinase (dAK/dGK) from Lactobacillus acidophilus R-26 by Ala, Asn, or Glu produced increased Km values for deoxyguanosine on dGK. However, it did not seem to affect the binding of Mg-ATP. The Asp-84 dGK replacements had no apparent effect on the binding of deoxyadenosine by dAK. However, the mutant dGKs were no longer inhibited by dGTP, normally a potent distal endproduct inhibitor of dGK. Moreover, the allosteric activation of dAK activity by dGTP or dGuo was lost in the modified heterodimeric dAK/dGK enzyme. Therefore, it seems very likely that Asp-84 participates in dGuo binding at the active site of the dGK subunit of dAK/dGK from Lactobacillus acidophilus R-26.

Allosteric Regulation↗

Development of probiotic cheese manufactured from goat milk: response surface analysis via technological manipulation.

Production of caprine milk has been rising steadily, partially because of its good nutritional value; the possibility of improving nutritional benefits by adding probiotic species such as Bifidobacterium lactis and Lactobacillus acidophilus was assessed. The manufacturing process of a traditional semi-hard goat cheese was technologically modified to optimize the process. The amount of starter inoculum, the concentration of salt, the addition of a protein hydrolysate, and the ripening time were varied to improve the microbiological, biochemical, and sensory properties of the cheese. Bifidobacterium lactis was able to grow slightly (up to 3 x 10(8) cfu/g), but growth was dependent on the physicochemical characteristics of the cheese. Lactobacillus acidophilus did not grow substantially in any of the experimental cheeses, and maximum numbers did not exceed 6 x 10(7) cfu/g. Concentrations of lactic acid and acetic acid increased throughout cheese manufacture, indicating that production of these acids was uncoupled from growth. Viability of the probiotic strains during ripening was sufficient to yield numbers that were above the accepted threshold (10(6) cfu/g) for a probiotic effect. Both strains contributed significantly to ripening, especially in the formation of low molecular mass peptides and amino acids, but lipolysis was not greatly affected. Statistical analyses using response surface methodology indicated that the manufacture of goat cheese could be optimized by the addition of 0.30% (vol/wt) milk hydrolysate, 3 x 10(7) of viable B. lactis and 7 x 10(6) of viable L. acidophilus cells/ml of milk, respectively, 3.50% (wt/wt) salt, and ripening for 70 d.

Acetic Acid↗

Evaluation of an avian-specific probiotic and Salmonella typhimurium-Specific antibodies on the colonization of Salmonella typhimurium in broilers.

Salmonella typhimurium colonizes the intestinal tract of poultry and causes food-borne illness in humans. Reduction of S. typhimurium colonization in the intestinal tract of poultry reduces potential carcass contamination during slaughter. The purpose of this study was to determine the effect of an avian-specific probiotic and S. typhimurium-specific antibodies on the colonization of S. typhimurium in broilers and on body weights. Broiler chicks were spray-vaccinated at the hatchery with the commercial product. Avian Pac Plus, which contains Lactobacillus acidophilus, Streptococcus faecium, ad S. typhimurium-specific antibodies. At placement, these chicks were administered Avian Pac plus in the water. Six hours postplacement, chicks were orally challenged with 1.8 x 10 (7) CFU of S. typhimurium. Chicks were administered Avian Pac Plus for two additional days postchallenge. Chicks were evaluated for S. typhimurium colonization and shedding every 3 to 4 days for the first 2 weeks and every 7 days for 6 weeks. The mean cecal and colonic concentration of S. typhimurium from the Avian Pac Plus-treated group was significantly lower at day 31 (P = 0.0001), day 38 (P = 0.0005), and day 43 (P = 0.0001) than the nontreated control group. These results indicated that a combination of Lactobacillus acidophilus, Streptococcus faecium, and S typhimurium-specific antibodies have a beneficial effect in reducing the colonization of S. typhimurium in market-aged broilers.

Abattoirs↗

Ingredient supplementation effects on viability of probiotic bacteria in yogurt.

The present investigation studied the effects of cysteine, whey powder, whey protein concentrate, acid casein hydrolysates, or tryptone on the viability of Streptococcus thermophilus, Lactobacillus acidophilus, and bifidobacteria. Changes in pH, titratable acidity, redox potential, and viability of bacteria were monitored during 24 h of fermentation and refrigerated storage (4 degrees C) of yogurt for 35 d. The incubation time that was needed to reach pH 4.5 was considerably affected by the added ingredients. Also, the drop in pH or the increase in acidity and redox potential was dependent on the added ingredients. The addition of cysteine, whey protein concentrate, acid casein hydrolysates, or tryptone improved the viability of bifidobacteria to a variable extent, but whey powder failed to improve their viability. The morphology of S. thermophilus, as shown by electron microscopy, was affected by cysteine at 500 mg/L, possibly as a result of reduced redox potential. Sodium dodecyl sulfate-PAGE and amino acid analyses suggested that the nitrogen source in the form of peptides and amino acids improved the viability of bifidobacteria in yogurt made with a commercial ABT (Lactobacillus acidophilus, bifidobacteria, and Streptococcus thermophilus) starter culture, which showed a dramatic decline in the counts of this organism in previous studies.

Bacteria↗

Evaluation of a diet containing probiotics and zinc for the treatment of mild diarrheal illness in children younger than one year of age.

OBJECTIVES: Supplementation of probiotics and supplementation of zinc during acute gastroenteritis in children have been shown to exert positive effects on diarrhea duration and severity. Our aim was to evaluate a new diet enriched with zinc and probiotic bacteria in the treatment of acute gastroenteritis in young children. METHODS: In a double blind prospective study, 65 children aged 6-12 months were randomized to receive 6 x 10(9) colony forming units of Streptococcus thermophilus, Bifidobacterium lactis, Lactobacillus acidophilus (2 x 10(9) of each strain), 10 mg of zinc/day, and 0.3 grams of fructo-oligosaccharides in the supplemented group (n = 33) or placebo (n = 32), given in a soy protein based rice cereal. For each child, age, sex, weight, degree of dehydration, the presence of fever or vomiting, stool frequency and consistency were recorded daily until diarrhea resolution. RESULTS: Diarrhea resolution occurred after 1.43 +/- 0.71 days in the supplemented group vs. 1.96 +/- 1.24 in the control group (p = 0.017). In the subset of children who presented with vomiting, time to vomiting resolution was 0.27 +/- 0.59 vs. 0.81 +/- 0.91 days in the supplemented and control groups, respectively (p = 0.06). On day 3, there was only 1 child with watery stools in the supplemented group versus 10 children in the control group (p = 0.02). CONCLUSIONS: In our series, the feeding of a cereal containing Streptococcus thermophilus, Bifidobacterium lactis, Lactobacillus acidophilus and zinc, reduced the severity and duration of acute gastroenteritis in young children. However, whether this combination is better than either the addition of probiotics or zinc alone is yet to be determined.

Acute Disease↗

Probiotic lactobacilli and bacteraemia in Stockholm.

The objective of the present study was to determine the incidence of lactobacilli-induced bacteraemia in Stockholm, Sweden, between January 1998 and March 2004, and to identify the possible presence of probiotic strains. Isolated strains were checked for growth on selective lactobacilli-agar and the biochemical profiles were determined. The lactobacilli strains were further analysed with molecular methodologies to ascertain eventual similarities with the probiotic strains Lactobacillus paracasei subsp. paracasei F19 (LMG P 17806), Lactobacillus acidophilus NCFB 1748 and Lactobacillus rhamnosus GG (ATCC 53103). The minimum inhibitory concentrations of selected antimicrobial agents were determined by the agar dilution method. The incidence of bacteraemia cases caused by lactobacilli remained at the same level during the study period and constituted to <1% of the total number of bacteraemia cases each y. L. paracasei subsp. paracasei F19, L. acidophilus NCFB 1748 and L. rhamnosus GG were not identified in any of the samples.

Anti-Bacterial Agents↗

Antimicrobial effect of acidified nitrate and nitrite on six common oral pathogens in vitro.

BACKGROUND: Salivary nitrate is positively correlated with plasma nitrate and its level is 9 times the plasma level after nitrate loading. Nitrate in saliva is known to be reduced to nitrite by oral bacteria. Nitrate and nitrite levels in saliva are 3 - 5 times those in serum in physiological conditions respectively in our previous study. The biological functions of high salivary nitrate and nitrite are still not well understood. The aim of this in vitro study was to investigate the antimicrobial effects of nitrate and nitrite on main oral pathogens under acidic conditions. METHODS: Six common oral pathogens including Streptococcus mutans NCTC 10449, Lactobacillus acidophilus ATCC 4646, Porphyromonas gingivalis ATCC 33277, Capnocytophaga gingivalis ATCC 33624, Fusobacterium nucleatum ATCC 10953, and Candida albicans ATCC 10231 were cultured in liquid medium. Sodium nitrate or sodium nitrite was added to the medium to final concentrations of 0, 0.5, 1, 2, and 10 mmol/L. All of the microorganisms were incubated for 24 to 48 hours. The optical densities (OD) of cell suspensions were determined and the cultures were transferred to solid nutrient broth medium to observe the minimum inhibitory concentration and minimum bactericidal/fungicidal concentration for the six tested pathogens. RESULTS: Nitrite at concentrations of 0.5 to 10 mmol/L had an inhibitory effect on all tested organisms at low pH values. The antimicrobial effect of nitrite increased with the acidity of the medium. Streptococcus mutans NCTC 10449 was highly sensitive to nitrite at low pH values. Lactobacillus acidophilus ATCC 4646 and Candida albicans ATCC 10231 were relatively resistant to acidified nitrite. Nitrate at the given concentrations and under acidic conditions had no inhibitory effect on the growth of any of the tested pathogens. CONCLUSION: Nitrite, at a concentration equal to that in human saliva, is both cytocidal and cytostatic to six principal oral pathogens in vitro, whereas nitrate at a similar concentration has no antimicrobial effect on these organisms.

Anti-Infective Agents↗

Effect of Lactobacillus isolates on the adhesion of pathogens to chicken intestinal mucus in vitro.

AIMS: The aims of this study were to investigate in vitro the effects of Lactobacillus isolates from a chicken on adhesion of pathogenic Salmonella and Escherichia coli to chicken intestinal mucus obtained from different intestinal regions. METHODS AND RESULTS: Bacteria were labelled by using methyl-1,2-[(3)H]-thymidine. The bacterial adhesion was assessed by measuring the radioactivity of bacteria adhered to the mucus. The results showed that the abilities of Lactobacillus spp. to bind to the same intestinal mucus were higher than those of pathogenic Salmonella and E. coli. Pretreatment of intestinal mucus with Lactobacillus fermentum and Lactobacillus acidophilus, alone or in combination, reduced the adhesion of the tested pathogens, but the reductive extent of pathogenic adhesion by Lactobacillus spp. in combination was relatively high. CONCLUSIONS: The tested bacteria had different adhesions to mucus glycoproteins isolated from different intestinal regions of chicken. Lactobacillus acidophilus and Lact. fermentum in combination revealed a better ability to inhibit attachments of Salmonella and E. coli to chicken intestinal mucus than Lactobacillus sp. alone. SIGNIFICANCE AND IMPACT OF THE STUDY: A mixture of intestinal Lactobacillus spp. from a chicken may play a protective role in excluding pathogenic Salmonella and E. coli from the intestine of chicken.

Animals↗

Factors involved in adherence of lactobacilli to human Caco-2 cells.

A quantitative assay performed with bacterial cells labelled with [3H]thymidine was used to investigate factors involved in the adherence of human isolates Lactobacillus acidophilus BG2FO4 and NCFM/N2 and Lactobacillus gasseri ADH to human Caco-2 intestinal cells. For all three strains, adherence was concentration dependent, greater at acidic pH values, and significantly greater than adherence of a control dairy isolate, Lactobacillus delbrueckii subsp. bulgaricus 1489. Adherence of L. acidophilus BG2FO4 and NCFM/N2 was decreased by protease treatment of the bacterial cells, whereas adherence of L. gasseri ADH either was not affected or was enhanced by protease treatment. Putative surface layer proteins were identified on L. acidophilus BG2FO4 and NCFM/N2 cells but were not involved in adherence. Periodate oxidation of bacterial cell surface carbohydrates significantly reduced adherence of L. gasseri ADH, moderately reduced adherence of L. acidophilus BG2FO4, and had no effect on adherence of L. acidophilus NCFM/N2. These results indicate that Lactobacillus species adhere to human intestinal cells via mechanisms which involve different combinations of carbohydrate and protein factors on the bacterial cell surface. The involvement of a secreted bridging protein, which has been proposed as the primary mediator of adherence of L. acidophilus BG2FO4 in spent culture supernatant (M.-H. Coconnier, T. R. Klaenhammer, S. Kernéis, M.-F. Bernet, and A. L. Servin, Appl. Environ. Microbiol. 58:2034-2039, 1992), was not confirmed in this study. Rather, a pH effect on Caco-2 cells contributed significantly to the adherence of this strain in spent culture supernatant.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Adhesion↗

Adherence of Lactobacillus species to human fetal intestinal cells.

Thirty-two lactobacilli were tested for ability to adhere to a human fetal intestinal epithelial cell line. By an in vitro system, two adherence mechanisms were found. One mechanism, requiring calcium in the adherence reaction, was nonspecific and allowed all lactobacilli tested to adhere. The other system, not requiring calcium, was found in four strains, all human Lactobacillus acidophilus isolates. Colonial morphology, serial broth passage, and exposure of cell crops to freezing or lyophilization did not affect adherence of Lactobacillus acidophilus. In vitro adherence, combined with subsequent in vivo studies, may provide a basis for screening candidate organisms for use in microbiotic supplements.

Adhesiveness↗

Does lactobacillus vaccine for trichomoniasis, Solco Trichovac, induce antibody reactive with Trichomonas vaginalis?

Solco Trichovac is a vaccine of Lactobacillus acidophilus developed for treating trichomoniasis. The efficacy of the vaccine is reportedly due to cross reacting antibody being produced in people immunised with the lactobacillus bacteria. Several techniques, including enzyme linked immunosorbent, indirect immunofluorescence, immunoblot, and radioimmunoprecipitation assays were used to assess the extent of antigenic cross reactivity between Lactobacillus acidophilus and Trichomonas vaginalis. Data show a lack of antigenic relatedness between the Solco Trichovac lactobacilli and several strains of T vaginalis. Furthermore, antiserum to L acidophilus failed to inhibit trichomonad cytadherence or host cell killing, as is suggested by the producers of the Solco Trichovac vaccine.

Animals↗

Biocatalysis of linoleic acid to conjugated linoleic acid.

CLA refers to a group of geometrical and positional isomers of linoleic acid (LA) with conjugated double bonds. CLA has been reported to have diverse health benefits and biological properties. Traditional organic synthesis is highly capital-intensive and results in an isomeric mixture of CLA isomers. Biotechnology presents new alternatives to traditional lipid manufacturing methods. The objective of this study was to examine the effect of protein isolation procedures on linoleate isomerase (LAI) recovery from microbial cells and biocatalysis of LA to CLA. Protein isolation experiments were carried out using Lactobacillus acidophilus L1 and two strains of Lactobacillus reuteri (ATCC 23272 and ATCC 55739). Under the same assay conditions, ATCC 55739 had the highest LAI activity among the microbial cultures examined in this study. Efficiency of cell lysis methods, which included various combinations of lysozyme and mutanolysin treatments in combination with sonication and osmotic rupture of cells with liquid nitrogen, was very low. Although treatment of cell material with a detergent (octylthioglucoyranoside) freed a significant amount of LAI activity into the solution, it was not sufficient to recover all the LAI activity from the residual cells. Crude LAI preparations produced mainly the cis-9,trans-11 CLA isomer. Time and substrate/protein ratio had a significant effect on biocatalysis of LA to CLA. It appears that the mechanism and kinetics of enzymatic conversion of LA to CLA are quite complex and requires further research using pure LAI preparations.

Lactobacillus acidophilus↗