Assay for biotin in the presence of dethiobiotin with Lactobacillus plantarum

At concentrations greater than approximately 0.5 microM, dethiobiotin can cause the bioassay for biotin, which employs Lactobacillus plantarum, to over value the actual biotin level. This can be as much as 30-fold at 10 microM DL-dethiobiotin and 5 pM biotin. Dethiobiotin does this by exerting a sparing effect on the biotin response by the assay organism. We demonstrate one way to determine the actual biotin concentration in the presence of interfering levels of dethiobiotin.     

Oxygen utilization by Lactobacillus plantarum

Cell-free extracts of Lactobacillus plantarum contain non-proteinaceous compounds which mimic superoxide dismutase activity. Using the test system in which O ₂ ^– is generated by xanthine oxidase, superoxide dismutase activity is found in cell-free extracts, where proteins are removed by precipitation. This activity is strongly decreased after dialysis of cell-free extracts. Superoxide dismutase activity was also investigated by means of pulse radiolysis. Cell-free extracts of Escherichia coli were also investigated as a comparison, which were known to contain superoxide dismutase. With cell-free extracts of both L. plantarum and E. coli the decay of O ₂ ^– was markedly increased. However, the type of reaction of the O ₂ ^– decay was of first order in the presence of E. coli extracts due to superoxide dismutase(s), and of second order in the presence of L. plantarum extracts, indicating that O ₂ ^– elimination is not an enzymic reaction. Mn²⁺ phosphate(s) might be responsible for the observed elimination of O ₂ ^– . The production of O ₂ ^– is not detectable during NADH-, lactate- or pyruvate oxidase reactions in L. plantarum extracts.     

Increased Sensitivity of Immunofluorescent Assay for Salmonella in Nonfat Dry Milk

The necessity of developing a quick, sensitive, and reliable test for Salmonella in nonfat dry milk (NDM) is evident from the recent tracing of Salmonella outbreaks to this product. Normally, coagulation of casein occurs when assaying NDM under regular cultural conditions, raising the possibility of trapped bacteria. After 20 hr of incubation of NDM in preenrichment lactose broth, enrichment was achieved by using Selenite-Cystine Broth. Smears from the enrichment broth were examined by the fluorescent-antibody technique (FAT) with a commercially available polyvalent O globulin conjugated with fluorescein. Standard cultural methods (SCM) were performed for comparison with FAT. Sensitivity of FAT was definitely improved by the use of trypsin. Casein coagulation of NDM can be avoided by addition of trypsin to samples during initial preenrichment in lactose broth. Samples containing approximately one Salmonella per 10 g were easily detected by FAT with the use of trypsin-treated samples. The method required only 42 hr to complete. Additionally, the use of trypsin enhanced recovery of Salmonella by use of SCM, as evidenced by alteration in the observed coliform to Salmonella ratios.     

Anaerobic l-lactate degradation by Lactobacillus plantarum

Abstract Lactobacillus plantarum strains used as silage inoculants were investigated for their ability to metabolize lactic acid anaerobically after prolonged incubation (7–30 days) when glucose was absent from the medium. When citrate was present in the medium together with glucose during the initial fermentation, the lactic acid produced was degraded. Citrate was concomitantly degraded, resulting in accumulation of formic, acetic and succinic acids along with CO₂. The anaerobic degradation was confirmed by the use of l ¹⁴C(U) labelled lactate. The existence of pyruvate formate lyase in L. plantarum was indicated by using ¹⁴C-labelled pyruvate and HPLC identification of end-products. The 1-¹⁴C-carboxylic acid group of pyruvate was converted to formic acid, and the 3-¹⁴C was found in acetic acid. The key enzyme(s) in this metabolic pathway appears to require anaerobic conditions and induction by citrate.     

植物乳杆菌细菌素的研究与应用

植物乳杆菌细菌素不仅种类多,产生菌在发酵过程中还可产生良好的保健功效,因此成为研究的热点。本文对植物乳杆菌细菌素的种类、分子结构、抑菌机制及遗传控制做了较为详尽的介绍,并简要介绍了植物乳杆菌细菌素在食品、医药、饲料中的应用,为进一步研究植物乳杆菌细菌素提供了参考。     

Identification of Antioxidants Produced by Lactobacillus plantarum

We identified two compounds that demonstrated 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity from cultures of Lactobacillus plantarum. Spectroscopic analyses proved these compounds to be L-3-(4-hydroxyphenyl) lactic acid (HPLA) and L-indole-3-lactic acid (ILA). The respective EC₅₀ values for HPLA and ILA were 36.6 ± 4.3 mM and 13.4 ± 1.0 mM.     

植物乳杆菌高活力保存方法的研究及其应用

本研究旨在探讨植物乳杆菌的高活力保存方法,为植物乳杆菌饲料添加剂规模化、工业化生产奠定基础。采用4℃低温保存法、36℃烘干后常温保存法、阳离子活性载体保存法等3种方法对植物乳杆菌进行活力保存比较试验。以保存后活菌数不低于原值50%为参照标准,结果显示,对照组可保存15 d,4℃低温保存法可保存30 d,36℃烘干后常温保存法只能保存一周,而阳离子活性载体保存法则可保存2个月以上。结果表明,阳离子活性载体保存法可应用于植物乳杆菌饲料添加剂的规模化、工业化生产实践中。     

Lactobacillus casei and Lactobacillus plantarum initiate catabolism of methionine by transamination

AIMS: To study the ability of Lactobacillus casei and Lact. plantarum strains to convert methonine to cheese flavour compounds. METHODS AND RESULTS: Strains were assayed for methionine aminotransferase and lyase activities, and amino acid decarboxylase activity. About 25% of the strains assayed showed methionine aminotransferase activity. The presence of glucose in the reaction mixture increased conversion of methionine to 4-methylthio-2-ketobutanoate (KMBA) and 4-methylthio-2-hydroxybutanoate (HMBA) in all strains. The methionine aminotransferase activity in Lact. plantarum and Lact. casei showed variable specificity for the amino group acceptors glyoxylate, ketoglutarate, oxaloacetate and pyruvate. None of the strains showed methionine lyase or glutamate and methionine decarboxylase activities. CONCLUSION: The presence of amino acid converting enzymes in lactobacilli is strain specific. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this work suggest that lactobacilli can be used as adjuncts for flavour formation in cheese manufacture.     

Oxygen dependent lactate utilization by Lactobacillus plantarum

Lactobacillus plantarum P5 grew aerobically in rich media at the expense of lactate; no growth was observed in the absence of aeration. The oxygen-dependent growth was accompanied by the conversion of lactate to acetate which accumulated in the growth medium. Utilization of oxygen with lactate as substrate was observed in buffered suspensions of washed whole cells and in cell-free extracts. A pathway which accounts for the generation of adenosine triphosphate during aerobic metabolism of lactate to acetate via pyruvate and acetyl phosphate is proposed. Each of the enzyme activities involved, nicotinamide adenine dinucleotide independent lactic dehydrogenase, nicotinamide adenine dinucleotide dependent lactic dehydrogenase, pyruvate oxidase, acetate kinase and NADH oxidase were demonstrated in cell-free extracts. The production of pyruvate, acetyl phosphate and acetate was demonstrated using cell-free extracts and cofactors for the enzymes of the proposed pathway.Abbreviations MRS Man, Rogosa and Sharpe (1960) medium modified as in Materials and methods - TY Tryptone Yeast Extract broth - OUL Oxygen uptake with lactate as substrate - DCPIP 2,6-Dichlorophenolindophenol - LDH Lactic dehydrogenase     

Improvement of the Microbiological Assay for Calcium Leucovorin

The calcium leucovorin assay was improved by adding THORAL germicide which resuspended the cells so that an accurate turbidity reading could be obtained.     

Culture conditions of tannase production by Lactobacillus plantarum

Lactobacillus plantarum produced an extracellular tannase after 24 h growth on minimal medium of amino acids containing 2 g tannic acid l^–1. Enzyme production (6 U ml^–1) was optimal at 37 °C and pH 6 with 2 g glucose l^–1 and 7 g tannic acid l^–1 in absence of O₂.     

Liquid Nitrogen Freezing in Microbiological Assay Systems: I. Preservation of Lactobacillus leichmannii for Direct Use in the Vitamin B12 Assay

Suspensions of Lactobacillus leichmannii were stored in liquid nitrogen and were used as direct inocula in vitamin B₁₂ assays. Complete recovery of viable cells was obtained when the suspensions in basal B₁₂ medium were rapidly frozen by direct immersion into liquid nitrogen and rapidly thawed by agitating the suspensions in a water bath at 40 C. Greater than 90% destruction occurred when the suspensions were in saline. However, both suspensions were usable in the B₁₂ assay system. Assay results on a number of test materials indicated good correlation between freshly prepared suspensions and frozen suspensions in basal medium stored 3 months. Suspensions in saline stored for 1 year in liquid nitrogen showed no detectable difference from the first day after freezing. Suspensions frozen slowly at the rate of 1 degree per min from 4 to -40 C and subsequently immersed in liquid nitrogen had a longer lag period of growth and were not usable in the 18-hr assay incubation system. A major advantage of a stored inoculum for direct use in a microbiological assay is the reduced day-to-day variation in the inoculum.     

Rapid Microbiological Assay for 5-Fluorocytosine

A microbiological assay was developed for 5-fluorocytosine based on inhibition by the drug of the fall in pH produced by Candida albicans in a synthetic liquid culture medium. Serum and urine levels of the drug could be measured reproducibly in four hours.     

Energy conservation in malolactic fermentation by Lactobacillus plantarum and Lactobacillus sake

A comparably poor growth medium containing 0.1% yeast extract as sole non-defined constituent was developed which allowed good reproducible growth of lactic acid bacteria. Of seven different strains of lactic acid bacteria tested, only Lactobacillus plantarum and Lactobacillus sake were found to catalyze stoichiometric conversion of l-malate to l-lactate and CO₂ concomitant with growth. The specific growth yield of malate fermentation to lactate at pH 5.0 was 2.0 g and 3.7 g per mol with L. plantarum and L. sake, respectively. Growth in batch cultures depended linearly on the malate concentration provided. Malate was decarboxylated nearly exclusively by the cytoplasmically localized malo-lactic enzyme. No other C₄-dicarboxylic acid-decarboxylating enzyme activity could be detected at significant activity in cell-free extracts. In pH-controlled continuous cultures, L. plantarum grew well with glucose as substrate, but not with malate. Addition of lactate to continuous cultures metabolizing glucose or malate decreased cell yields significantly. These results indicate that malo-lactic fermentation by these bacteria can be coupled with energy conservation, and that membrane energetization and ATP synthesis through this metabolic activity are due to malate uptake and/or lactate excretion rather than to an ion-translocating decarboxylase enzyme.