Inhibitory effect of polyphenols from sumac, pomegranate and Indian almond on urease producing bacteria and jack bean urease activity.

Urinary tract infection caused by Klebsiella, Proteus and Streptococcus is a urease dependent process, hence treatment of these infections by antibacterial compounds lies in inhibition of their virulence factors. The crude methanolic extracts derived from sumac fruit, pomegranate peel and Indian almond leaves were separated into anthocyanin and non-anthocyanin fractions using solid phase cartridges. The inhibitory effect of these fractions was determined on the growth of urease producing species and jack bean urease activity. Known compounds in the fractions were also docked with ureases of different biological origins viz. K. pneumoniae (PDB ID: 8HCN), K. aerogenes (PDB ID: 2KAU), Helicobacter pylori (PDB ID:8HC1)and Canavalia ensiformis (jack bean) (PDB ID: 3LA4) to determine their binding affinities and interaction with the enzyme. All the fractions showed significant inhibition growth for P. mirabilis, S. epidermidis and K.pneumoniae. Among the samples, sumac showed greatest inhibition against all (MIC 6-25 mg.mL-1) while among the fractions, anthocyanin was found to be most active (MIC 6-12 mg/mL). Likewise, all fractions inhibited urease with lowest ICs50 shown by sumac fractions (21-116 µg.mL-1). Out of 39 compounds docked, 27 showed interaction with movable flaps and/or active site of ureases which explains their mode of inhibition.

Antibacterial and antioxidant potential of the phenolic extract and its fractions isolated from Allium ascalonicum (onion) peel.

Aqueous methanolic (20:80) crude extract of the outer peel of Allium ascalonicum (onions) was fractionated into anthocyanin (II), acidic (i.e., phenolic acids, Ia), neutral A (i.e., flavanols, Ib) and neutral B (i.e., flavonols, Ic) phenols using C-18 and hydrophilic-lipophilic balanced (HLB) Co-polymer cartridges. The fractions were analysed for total phenolic, total flavonoids, antibacterial activity, antioxidant potential and lipoxygenase (LOX) inhibition. The recovery of total phenols and bio functional activity in all fractions were found to be more in HLB. All fractions showed antimicrobial activity against tested strains with phenolic acids (Ia) being most active and flavanol (Ib) least active, while the highest free radical scavenging, reducing power and LOX inhibition was exhibited by flavonol (Ic) and least by flavanols (Ib). HPLC analysis revealed the presence of vanillic acid and quercetin in non-anthocyanin I and cyanidin in anthocyanin II.

Binding effect of proline-rich-proteins (PRPs) on in vitro antimicrobial activity of the flavonoids.

The interaction of the cyanidin, pelargonidin, catechin, myrecetin and kaempferol with casein and gelatin, as proline rich proteins (PRPs), was studied. The binding constants calculated for both flavonoid-casein and flavonoid-gelatin were fairly large (10 (5) -10 (7) M (-1) ) indicating strong interaction. Due to higher proline content in gelatin, the binding constants of flavonoid-gelatin (2.5 × 10 (5) -6.2 × 10 (7) M (-1) ) were found to be higher than flavonoid-casein (1.2 × 10 (5) -5.0 × 10 (7) M (-1) ). All the flavonoids showed significant antibacterial activity against the tested strains. Significant loss in activity was observed due to the complexation with PRPs confirming that binding effectively reduced the concentration of the free flavonoids to be available for antibacterial activity. The decline in activity was corresponding to the values of the binding constants. Though the activities of free catechin and myrecetin were higher compared to pelargonidin, cyanidin and kaempferol yet the decline in activity of catechin and myrecetin due to complexation with casein and gelatin was more pronounced.

Binding effect of proline-rich-proteins (PRPs) on in vitro antimicrobial activity of the flavonoids

The interaction of the cyanidin, pelargonidin, catechin, myrecetin and kaempferol with casein and gelatin, as proline rich proteins (PRPs), was studied. The binding constants calculated for both flavonoid-casein and flavonoid-gelatin were fairly large (105–107 M−1) indicating strong interaction. Due to higher proline content in gelatin, the binding constants of flavonoid-gelatin (2.5 × 105–6.2 × 107 M−1) were found to be higher than flavonoid-casein (1.2 × 105–5.0 × 107 M−1). All the flavonoids showed significant antibacterial activity against the tested strains. Significant loss in activity was observed due to the complexation with PRPs confirming that binding effectively reduced the concentration of the free flavonoids to be available for antibacterial activity. The decline in activity was corresponding to the values of the binding constants. Though the activities of free catechin and myrecetin were higher compared to pelargonidin, cyanidin and kaempferol yet the decline in activity of catechin and myrecetin due to complexation with casein and gelatin was more pronounced.

Antimicrobial polyphenols from small tropical fruits, tea and spice oilseeds.

The polyphenolic fractions of fruits: Terminalia catappa, Carissa carandas, Ziziphus nummularia; spice oilseeds: thymol, mustard, fenugreek and poppy seeds; and herb: green and black teas were analyzed for their total phenolics, flavonoids and antimicrobial potential. All fractions from fruits, except anthocyanin of C. carandas, displayed substantial antibacterial activity in accordance to their phenolic contents, the difference in activity being quite significant (p < 0.05), highest for T. catappa (minimum inhibitory concentration, MIC: 7.8-1000 microg/mL) and lowest for C. carandas (MIC: 62.5-1000 microg/mL). With few exceptions, both green and black teas' fractions inhibited the tested strains, however, green tea fractions (MIC: 15.63-125 microg/mL) were more active than black (MIC: 31.25-1000 microg/mL) and neutral were more active than their corresponding acidic fractions. Oil fractions of all oilseeds were found to be more active than their polyphenolic fractions, their antibacterial action decreased in the order thymol > mustard > fenugreek > poppy seeds (p < 0.05). Though the fruits used for the study are underutilized and have been emphasized for processed products, they may potentially be important to fight against pathogenic bacteria in view of their MICs. The teas and oilseeds, though a small part of total food intake, are more functional and active against the tested bacterial species and may find potential applications in therapeutics and food preservation.

Antioxidant Potential of the Extracts, Fractions and Oils Derived from Oilseeds.

The polyphenolic extracts and oils were obtained from ajwain, mustard, fenugreek and poppy seeds. The extracts were partitioned into acidic and neutral polyphenolic fractions and following estimation of total phenolics in the crude extract, acidic and neutral fractions and oil, all were analyzed for their DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging potential, ferric reducing ability and chelating power. The highest amount of polyphenols was found in ajwain (8330 ± 107), then in mustard seeds (2844 ± 56.00) and in fenugreek (1130 ± 29.00), and least in poppy seeds (937 ± 18.52). The higher amounts of polyphenols were estimated in neutral fraction compared to acidic (p < 0.05). % Inhibition of DPPH by the crude extract and fractions of all oilseeds was quite significant, being higher for acidic than neutral. The highest % DPPH inhibition was shown by ajwain extract than mustard > fenugreek and least by poppy seed extracts (p < 0.05). The reducing power and the chelating effect of the oilseeds followed the same order as DPPH, but higher % chelation was shown by neutral than acidic fraction (p < 0.05). Though low in polyphenols, the oil fractions were as strong antioxidants as the acidic one. Though oilseeds are used in very small quantity in food, they are potential sources of natural antioxidants and may replace synthetic ones.

Epigallocatechin-3-gallate inhibits lactase but is alleviated by salivary proline-rich proteins.

Lactase phlorizin hydrolase is a small intestinal brush border enzyme that catalyzes the hydrolysis of the milk sugar, lactose, and also many flavonoid glucosides. We demonstrate that epigallocatechin-3-gallate (EGCG), the principal flavonoid from green tea, inhibits in vitro hydrolysis of lactose by intestinal lactase. We then tested the hypothesis that salivary proline-rich proteins (PRPs) could modulate this inhibition and stabilize EGCG. Inhibition by EGCG of digestive enzymes (α-amylase>chymotrypsin>trypsin>lactase≫pepsin) was alleviated ∼2-6-fold by PRPs. Furthermore, PRPs appeared stable to proteolysis and also stabilized EGCG under digestive conditions in vitro. This is the first report on EGCG inhibition of lactase, and it quantifies the protective role of PRPs against EGCG inhibition of digestive enzymes.

Antibacterial activity directed isolation of compounds from Onosma hispidum.

The chemical investigation of the ethanolic extract of the root bark of Onosma hispidum following antibacterial activity directed isolation led to the isolation of 4-hydroxy-3-methoxy cinnamic acid (ferulic acid) and 4-hydroxy-3-methoxy benzoic acid (vanillic acid) which have been reported for the first time in this species. In addition to these compounds, the crude ethanolic extract and methanol fraction exhibited substantial bioactivity against species of corynebacteria, enterococci, staphylococci and streptococci. Ferulic acid was found more bioactive (being relatively more hydrophobic) compared to vanillic acid.