Production of 3,4-dihydroxybenzoic acid by Azomonas macrocytogenes and Azotobacter paspali

1987 ◽  
Vol 33 (2) ◽  
pp. 169-175 ◽  
Author(s):  
S. Karen Collinson ◽  
James L. Doran ◽  
William J. Page
Keyword(s):  
Phenolic Compound ◽  
Protocatechuic Acid ◽  
Culture Supernatant ◽  
Carbon Sources ◽  
Defined Medium ◽  
Iron Binding ◽  
Free Medium ◽  
Dihydroxybenzoic Acid ◽  
Natural Iron ◽  
Cell Free Culture Supernatant

Four strains of Azomonas macrocytogenes and a strain of Azotobacter paspali were found to produce a common extracellular, iron-binding, phenolic compound. This compound was purified from cell-free culture supernatant fluids and chemically identified as 3,4-dihydroxybenzoic acid (protocatechuic acid). 3,4-Dihydroxybenzoic acid was produced by cells growing in a simple defined medium containing acetate and glucose or sucrose as sole carbon sources. This compound promoted the solubilization of iron from the minerals olivine, glauconite, pyrite, and marcasite. Mineral-free medium prepared by preincubation with olivine or glauconite as the only iron source supported enhanced growth yields of Azomonas macrocytogenes (ATCC 12334) when 3,4-dihydroxybenzoic acid was present during the preincubation period. This suggests that 3,4-dihydroxybenzoic acid may play a role in solubilizing iron from some natural iron sources; however, 3,4-dihydroxybenzoic acid can not be considered a true siderophore since it was produced by Azomonas macrocytogenes and Azotobacter paspali growing in iron-limited and iron-sufficient medium.

Growth and hemolysin production by Haemophilus pleuropneumoniae cultivated in a chemically defined medium

1986 ◽  
Vol 32 (10) ◽  
pp. 801-805 ◽  
Author(s):  
James R. Maudsley ◽  
Solomon Kadis
Keyword(s):  
Growth Rate ◽  
Yeast Extract ◽  
Hemolytic Activity ◽  
Culture Supernatant ◽  
Iron Concentration ◽  
Defined Medium ◽  
Chemically Defined Medium ◽  
Sterile Cell ◽  
Stages Of Growth ◽  
Cell Free Culture Supernatant

A chemically defined medium (CDM) has been developed which supports both growth and hemolysin production by Haemophilus pleuropneumoniae. Although the growth rate in stationary cultures was substantially slower in CDM than in trypticase soy broth plus 0.6% yeast extract (TSBYE) and slightly slower than in heart infusion broth (HIB), extracellular hemolysin activity in CDM was slightly higher than in HIB and 16-fold greater than in TSBYE. Maximum hemolytic activity was produced in CDM in early to mid log phase of growth. Hemolytic activity in sterile, cell-free culture supernatant fluids persisted for over 10 days at 4 °C and 3–5 days at 37 °C, but was completely destroyed at 56 °C after 30 min. Total hemolysin inactivation was also achieved in the presence of trypsin or pronase (10 units/mL), but no decrease in hemolytic activity was noted in the presence of DNase or RNase. Iron had little effect on the hemolytic activity in the early stages of growth. However, in the later stages of growth, iron had a pronounced effect with hemolyic activity decreasing as the iron concentration increased from 1 to 500 μM. None of these iron concentrations had any effect on the hemolytic activity when added directly to prepared cell-free culture supernatant fluids. The extracellur hemolysin produced by H. pleuropneumoniae in CDM appears to be a heat-labile protein the activity of which is influenced by iron at certain phases of growth.

scholarly journals Production of biologically active human interleukin-10 by Bifidobacterium bifidum BGN4

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Nayoun Hong ◽  
Seockmo Ku ◽  
Kyungjin Yuk ◽  
Tony V. Johnston ◽  
Geun Eog Ji ◽  
...  
Keyword(s):  
Culture Supernatant ◽  
Shuttle Vector ◽  
Sodium Dodecyl ◽  
Interleukin 10 ◽  
Biologically Active ◽  
Enzyme Linked Immunosorbent Assay ◽  
Functional Evaluation ◽  
Ht 29 ◽  
Cell Free Culture Supernatant

Abstract Background Bifidobacterium spp. are representative probiotics that play an important role in the health of their hosts. Among various Bifidobacterium spp., B. bifidum BGN4 exhibits relatively high cell adhesion to colonic cells and has been reported to have various in vivo and in vitro bio functionalities (e.g., anti-allergic effect, anti-cancer effect, and modulatory effects on immune cells). Interleukin-10 (IL-10) has emerged as a major suppressor of immune response in macrophages and other antigen presenting cells and plays an essential role in the regulation and resolution of inflammation. In this study, recombinant B. bifidum BGN4 [pBESIL10] was developed to deliver human IL-10 effectively to the intestines. Results The vector pBESIL10 was constructed by cloning the human IL-10 gene under a gap promoter and signal peptide from Bifidobacterium spp. into the E. coli-Bifidobacterium shuttle vector pBES2. The secreted human IL-10 from B. bifidum BGN4 [pBESIL10] was analyzed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), Western Blotting, and enzyme-linked immunosorbent assay (ELISA). More than 1,473 ± 300 ng/mL (n = 4) of human IL-10 was obtained in the cell free culture supernatant of B. bifidum BGN4 [pBESIL10]. This productivity is significantly higher than other previously reported human IL-10 level from food grade bacteria. In vitro functional evaluation of the cell free culture supernatant of B. bifidum BGN4 [pBESIL10] revealed significantly inhibited interleukin-6 (IL-6) production in lipopolysaccharide (LPS)-induced Raw 264.7 cells (n = 6, p < 0.0001) and interleukin-8 (IL-8) production in LPS-induced HT-29 cells (n = 6, p < 0.01) or TNFα-induced HT-29 cells (n = 6, p < 0.001). Conclusion B. bifidum BGN4 [pBESIL10] efficiently produces and secretes significant amounts of biologically active human IL-10. The human IL-10 production level in this study is the highest of all human IL-10 production reported to date. Further research should be pursued to evaluate B. bifidum BGN4 [pBESIL10] producing IL-10 as a treatment for various inflammation-related diseases, including inflammatory bowel disease, rheumatoid arthritis, allergic asthma, and cancer immunotherapy.

Anti-tumor potential of cell free culture supernatant of Lactobacillus rhamnosus strains isolated from human breast milk

2019 ◽  
Vol 123 ◽  
pp. 286-297 ◽  
Author(s):  
Muhammad Shahid Riaz Rajoka ◽  
Haobin Zhao ◽  
Hafiza Mahreen Mehwish ◽  
Na Li ◽  
Yao Lu ◽  
...  
Keyword(s):  
Breast Milk ◽  
Culture Supernatant ◽  
Lactobacillus Rhamnosus ◽  
Human Breast Milk ◽  
Human Breast ◽  
Cell Free Culture Supernatant

scholarly journals The growth of Paracoccus halodenitrificans in a defined medium

1984 ◽  
Vol 30 (6) ◽  
pp. 837-840 ◽  
Author(s):  
Lawrence I. Hochstein ◽  
Geraldine A. Tomlinson
Keyword(s):  
Synthetic Medium ◽  
Carbon Sources ◽  
Defined Medium ◽  
Anaerobic Growth ◽  
Inorganic Salts ◽  
Vitamin B ◽  
Aerobic Growth ◽  
Methionine Biosynthesis ◽  
Dependent Pathway

A synthetic medium, consisting of inorganic salts and any of a number of carbon sources, supported the aerobic growth of Paracoccus halodenitrificans when supplemented with thiamine. The same medium plus an appropriate nitrogenous oxide supported anaerobic growth when additionally supplemented with methionine. The observation that vitamin B12 or betaine replaced methionine suggested that P. halodenitrificans had a defect in the cobalamin-dependent pathway for methionine biosynthesis, as well as the inability to synthesize betaine when growing anaerobically.

Purine metabolism in Neisseria gonorrhoeae: the requirement for hypoxanthine

1980 ◽  
Vol 26 (1) ◽  
pp. 13-20 ◽  
Author(s):  
Stephen A. Morse ◽  
Lynne Bartenstein
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Growth Medium ◽  
De Novo ◽  
Defined Medium ◽  
Purine Metabolism ◽  
Purine Biosynthesis ◽  
Free Medium ◽  
De Novo Purine Biosynthesis ◽  
Reduced Rate

Strains isolated from disseminated gonococcal infections often require hypoxanthine for growth. The biochemical bases for the requirement for hypoxanthine in strains isolated from both disseminated (Ile−Val−Arg−Hyx−Ura−phenotype) and non-disseminated (Hyx−phenotype) infections were compared. The requirement for hypoxanthine was dependent upon the composition of the growth medium. In a complete defined medium, hypoxanthine was replaced by a mixture of adenine and guanine but not by either purine alone. The addition of adenine alone inhibited gonococcal growth. This inhibition was reversed by the addition of guanine and most likely resulted from an inhibition of de novo purine biosynthesis. In a histidine-free medium, adenine replaced the hypoxanthine requirement in Ile−Val−Arg−Hyx−Ura− strains. Adenine did not replace the hypoxanthine requirement in Hyx− strains. The Ile−Val−Arg−Hyx−Ura− strains exhibited a markedly reduced rate of de novo purine biosynthesis while Hyx− strains were blocked in this pathway. In vivo concentrations of purines are important factors which may limit the intracellular or extracellular growth of these strains.

Heat‐killed Lactobacillus reuteri and cell‐free culture supernatant have similar effects to viable probiotics during interaction with Porphyromonas gingivalis

2020 ◽  
Vol 55 (2) ◽  
pp. 215-220 ◽  
Author(s):  
Barbara M. C. Geraldo ◽  
Marianna N. Batalha ◽  
Noala V. M. Milhan ◽  
Rodnei D. Rossoni ◽  
Liliana Scorzoni ◽  
...  
Keyword(s):  
Porphyromonas Gingivalis ◽  
Culture Supernatant ◽  
Lactobacillus Reuteri ◽  
Cell Free Culture Supernatant

Changes in Heat Resistance Resulting from pH and Nutritional Shifts of Acid-Adapted and Non–Acid-Adapted Listeria monocytogenes Scott A†

2004 ◽  
Vol 67 (2) ◽  
pp. 316-321 ◽  
Author(s):  
DARRELL O. BAYLES
Keyword(s):  
Listeria Monocytogenes ◽  
Heat Resistance ◽  
Frequency Distribution ◽  
Culture Supernatant ◽  
Physiological State ◽  
Distribution Data ◽  
Treatment Conditions ◽  
Lower Heat ◽  
Full Factorial ◽  
Cell Free Culture Supernatant

Stationary-phase Listeria monocytogenes cells that were either pH dependent acid adapted or not acid adapted were heat challenged at 60°C in a two-level full factorial design for three variables. The three variables and the levels consisted of tryptic soy broth (TSB) and sterile cell-free culture supernatant (sterile TSB), the presence and absence of 1% added glucose, and pH 4.8 and pH 7. Non–acid-adapted cells were most heat resistant when challenged in TSB (mean decimal reduction times at 60°C: D60 = 1.16 min). In the absence of added glucose, non–acid-adapted cells had similar D60-values for inactivations at pH 4.8 and pH 7; however, the presence of glucose caused non–acid-adapted cells challenged at pH 4.8 to be more heat sensitive (D60 = 0.65 min) than those inactivated at pH 7 (D60 = 1.03 min), indicating an interaction between glucose and pH. Overall, the significantly decreased heat resistance of the acid-adapted cells was due to the presence of glucose (D60 = 0.78 min without glucose, D60 = 0.59 min with glucose). Acid-adapted cells heat challenged in TSB had similar D60-values for inactivations at pH 4.8 and pH 7; however, acid-adapted cells in sterile TSB challenged at pH 4.8 (D60 = 0.52 min) had significantly lower heat resistance than did cells challenged at pH 7 (D60 = 0.76 min), indicating an interaction between the medium and pH. The L. monocytogenes survivor data were modeled to extract information on the frequency distribution of heat resistance within heat-challenged populations, and the frequency distribution characteristics of mean, mode, and variance were compared among treatment conditions. Significant differences in the frequency distribution data were compared with the D60-values. These data indicated that the presence and level of cross-protection is highly dependent on the physiological state of the cells and nutrient availability at the time of heat challenge. Such conditions should be considered to ensure that stressed pathogens in foods are destroyed or inactivated.

scholarly journals Antioxidant Activity of Pine Bark Constituents

2003 ◽  
Vol 58 (5-6) ◽  
pp. 351-354 ◽  
Author(s):  
Ammar Saleem ◽  
Henri Kivelä
Keyword(s):  
Antioxidant Activity ◽  
Plant Material ◽  
Protocatechuic Acid ◽  
Pine Bark ◽  
Bark Extract ◽  
Inhibition Assay ◽  
Dihydroxybenzoic Acid ◽  
Lipid Peroxidation Inhibition

AbstractA modified in vitro lipid peroxidation inhibition assay was used to guide the fractionation and the isolation of antioxidative principles of Finnish pine bark extract. This approach yielded 3,4-dihydroxybenzoic acid (protocatechuic acid) and taxifolin-3-O-β-glucopyranoside as major antioxidative compounds from the plant material. The structural elucidation of these compounds was undertaken with the help of HPLC-DAD and HPLC-ESI-MS analyses. Their IC50 values, in comparison to trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), were: trolox (1.78 ± 0.56 μm) < protocatechuic acid (5.77 ± 1.63 μm) < taxifolin-3- O-β-glucopyranoside (16.30 ± 1.98 μm). The method for the determination of antioxidant activity proved reproducible and quick for routine analyses with 96 well plates.

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