Hydrophobic Fatty Acid Chains Research Articles

Sustainable biosurfactant production from secondary feedstock-recent advances, process optimization and perspectives.

Biosurfactants have garnered increased attention lately due to their superiority of their properties over fossil-derived counterparts. While the cost of production remains a significant hurdle to surpass synthetic surfactants, biosurfactants have been anticipated to gain a larger market share in the coming decades. Among these, glycolipids, a type of low-molecular-weight biosurfactant, stand out for their efficacy in reducing surface and interfacial tension, which made them highly sought-after for various surfactant-related applications. Glycolipids are composed of hydrophilic carbohydrate moieties linked to hydrophobic fatty acid chains through ester bonds that mainly include rhamnolipids, trehalose lipids, sophorolipids, and mannosylerythritol lipids. This review highlights the current landscape of glycolipids and covers specific glycolipid productivity and the diverse range of products found in the global market. Applications such as bioremediation, food processing, petroleum refining, biomedical uses, and increasing agriculture output have been discussed. Additionally, the latest advancements in production cost reduction for glycolipid and the challenges of utilizing second-generation feedstocks for sustainable production are also thoroughly examined. Overall, this review proposes a balance between environmental advantages, economic viability, and societal benefits through the optimized integration of secondary feedstocks in biosurfactant production.

Membranes and Transport Mechanisms

Cellular life depends on the movement of membrane ions. For cells to continue functioning as they go through their life cycle, there must be a tremendous amount of exchange. Cell membranes are active structures rather than merely passive barriers separating cells from their surroundings. Cell membranes are known to be selectively permeable, allowing some chemicals to pass through while preventing others from doing so. Therefore, all cellular exchange activities can have active participation from cell membranes. Through the membrane interface, elements needed for cellular existence, communication among cells, and interaction between cells and their environment are taken in. Additionally, cells have the ability to direct and influence input and export processes, as well as moderate the subsequent signal response. Effective drug transportation across cellular membranes is essential for all treatment procedures. The majority of medications used in cancer therapy require carrier proteins for their transmembrane transport, even though hydrophobic small molecules can enter the cell membrane through straightforward diffusion. As a result, many scientists concentrate on creating novel strategies to improve drug delivery and uptake into desired cells. A live cell's plasma membrane serves as its outer wall. Although it acts as a barrier, it also permits transport between the cell and its surroundings.
 While the hydrophilic head groups are in contact with the surrounding water, the bilayer's hydrophobic interior, which is made up of fatty acid chains, renders the membrane impermeable to water-soluble compounds. To reduce the accessibility of hydrophobic fatty acid chains, phospholipids can spontaneously form stable bilayers in an aqueous environment due to their amphipathic nature. As lipids and proteins positioned in the plasma membrane can rotate or diffuse laterally and are distributed asymmetrically in the two leaflets, membranes are extremely dynamic structures. In addition to glycolipids, the outer leaflet of the plasma membrane mostly consists of the phospholipids phosphatidylcholine and sphingomyelin. Phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol are the three main substances that make up the inner leaflet.
 Additionally, both leaflets contain cholesterol, which affects the fluidity of the membrane. Lipid rafts are particular domains abundant in sphingolipids and cholesterol. The plasma membrane's basic role as a selectively permeable wall dividing the inside from the outside of the cell is determined by the structure of the phospholipid bilayer. Membrane proteins are essential for guaranteeing the selective translocation of molecules across the membrane as well as the regulation of cellular connections. They can function as energy converters, ion channels, pumps, receptors, or enzymes. The maintenance of the osmotic pressure and cellular pH, as well as the regulation of drug uptake mechanisms, account for the regulation of fundamental cellular activities.

Peptide amphiphile inspired self-assembled, ordered gold nanocomposites for improved sensitivity of electrochemical immunosensor: Applications in determining the total aflatoxin amount in food stuffs

In peptide amphiphile, The positively charged amino acid arginine can inspire the ordered self-assembly of gold nanocomposites (AuNPs), transfer positive charge to AuNPs, and weaken the aggregation of AuNPs by electrostatic repulsion, whereas hydrophobic fatty acid chains regulate the self-assembly of AuNPs through hydrophobic interaction, which may be a novel strategy to overcome disordered arrangement and aggregation of AuNPs to obtain an ultra-sensitive electrochemical immunosensor for determining the total aflatoxin amount. In this study, a peptide amphiphile (C14R5), composed of five arginine residues as the hydrophilic chain and myristic acid as the hydrophobic chain, inspired AuNPs to form monodispersed hollow raspberry-like AuNPs (rasAuNPs). rasAuNPs could captured and immobilized large amounts of aflatoxin antigens via the Au–S bonds, resulting in binding to more anti-aflatoxin antibodies. In the absence of aflatoxins, the enriched antigens bound to abundant antibodies, resulting in a low blank signal current. By contrast, in the presence of aflatoxins, enough antibodies could bind to the targets and less antibodies could recognize the antigens, increasing the detection signal intensity. Under the optimal conditions, the developed sensor demonstrated a wide linear range (0.13–29.06 pg mL−1) and a low limit of detection for total aflatoxins (0.05 pg mL−1) using a mixed standard (AFB1: AFB2: AFG1: AFG2 with a weight ratio of 1:1:1:1) in peanut, peanut milk, and maize powder samples. Hence, this novel strategy improves the sensitivity of electrochemical sensors and can be easily applied to detect other small molecule compound for the purpose of food safety.

Hemicellulose-based nanocomposites coating delays lignification of green asparagus by introducing AKD as a hydrophobic modifier

Fresh green asparagus is prone to lignify after harvest due to its high respiration rate and secretion of secondary metabolites regulated by enzymes generated during storage. In this work, a dense hemicellulose-based coating containing hemicellulose, cellulose nanofibers, and montmorillonite (MMT) was prepared and used as a coating to reduce the respiration rate of green asparagus, with which the shelf life of green asparagus was prolonged from 5 days to 7 days. Alkyl ketene dimer (AKD) was used as a hydrophobizer and a crosslinking agent to reduce humidity sensitivity by introducing hydrophobic fatty acid chains to nanoparticles for increased structural stability and respiratory barrier property hemicellulose-based film in high humidity conditions. Our study proved that AKD modified coating with nature-derived nanomaterials could effectively increase respiratory barrier property, slow down the secretion of lignification-involved enzymes, and regulating the conversion of phenolic compounds to the lignin monomers.

Purification of surfactin compounds produced by a Bacillus subtilis strain

Surfactins are lipopeptide-type biosurfactants produced mainly by Bacillus species containing a peptide loop of seven amino acids and a hydrophobic fatty acid chain. These molecules exhibit various biological activities; therefore, their therapeutic and environmental applications are in the focus worldwide. In our work, a multi-step purification and separation process was developed to isolate surfactins from the ferment broth of B. subtilis SZMC 6179J strain. The process incorporates normal phase flash chromatography for pre-purifying the crude extract and two consecutive reverse phase HPLC separations for the isolation of the various surfactin molecules. The determination of the relative amounts of lipopeptides both in the crude extract and in each fraction of every separation step were carried out by HPLC-HESI-MS examinations. The ratio of surfactins in the crude extract was 21.35%, but after the preparative flash chromatographic separation the relative amount of surfactins was observed to be 30.44%. The preparative HPLC purification step resulted 85.39% purity of the surfactins. Nine different surfactin variants were isolated and identified from the fractions of this final semi-preparative HPLC purification, out of which three compounds were completely purified, and three others were detected in relative amounts of more than 95% in some fractions.

Lysophospholipids in Lung Inflammatory Diseases.

The lysophospholipids (LPLs) belong to a group of bioactive lipids that play pivotal roles in several physiological and pathological processes. LPLs are derivatives of phospholipids and consist of a single hydrophobic fatty acid chain, a hydrophilic head, and a phosphate group with or without a large molecule attached. Among the LPLs, lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are the simplest, and have been shown to be involved in lung inflammatory symptoms and diseases such as acute lung injury, asthma, and chronic obstructive pulmonary diseases. G protein-coupled receptors (GPCRs) mediate LPA and S1P signaling. In this chapter, we will discuss on the role of LPA, S1P, their metabolizing enzymes, inhibitors or agonists of their receptors, and their GPCR-mediated signaling in lung inflammatory symptoms and diseases, focusing specially on acute respiratory distress syndrome, asthma, and chronic obstructive pulmonary disease.

Ultrashort Cationic Lipopeptides-Effect of N-Terminal Amino Acid and Fatty Acid Type on Antimicrobial Activity and Hemolysis.

Ultrashort cationic lipopeptides (USCLs) are promising antimicrobial agents that hypothetically may be alternatively used to combat pathogens such as bacteria and fungi. In general, USCLs consist of fatty acid chains and a few basic amino acid residues. The main shortcoming of USCLs is their relatively high cytotoxicity and hemolytic activity. This study focuses on the impact of the hydrophobic fatty acid chain, on both antimicrobial and hemolytic activities. To learn more about this region, a series of USCLs with different straight-chain fatty acids (C8, C10, C12, C14) attached to the tripeptide with two arginine residues were synthesized. The amino acid at the N-terminal position was exchanged for proteinogenic and non-proteinogenic amino acid residues (24 in total). Moreover, the branched fatty acid residues were conjugated to N-terminus of a dipeptide with two arginine residues. All USCLs had C-terminal amides. USCLs were tested against reference bacterial strains (including ESKAPE group) and Candida albicans. The hemolytic potential was tested on human erythrocytes. Hydrophobicity of the compounds was evaluated by RP-HPLC. Shortening of the fatty acid chain and simultaneous addition of amino acid residue at N-terminus were expected to result in more selective and active compounds than those of the reference lipopeptides with similar lipophilicity. Hypothetically, this approach would also be beneficial to other antimicrobial peptides where N-lipidation strategy was used to improve their biological characteristics.

Effects of Different Cultivation Parameters on the Production of Surfactin Variants by a Bacillus subtilis Strain.

Surfactins are lipopeptide-type biosurfactants produced mainly by Bacillus species, consisting of a peptide loop of seven amino acids and a hydrophobic fatty acid chain (C12–C16). These molecules have been proven to exhibit various biological activities; thus, their therapeutic and environmental applications are considered. Within the surfactin lipopeptide family, there is a wide spectrum of different homologues and isomers; to date, more than 30 variants have been described. Since the newest members of these lipopeptides were described recently, there is no information that is available on their characteristic features, e.g., the dependence of their production from different cultivation parameters. This study examined the effects of both the different carbon sources and various metal ions on the surfactin production of a selected B. subtilis strain. Among the applied carbon sources, fructose and xylose had the highest impacts on the ratio of the different variants, regarding both the peptide sequences and the lengths of the fatty acids. Furthermore, the application of metal ions Mn2+, Cu2+ and Ni2+ in the media completely changed the surfactin variant compositions of the fermenting broths leading to the appearance of methyl esterified surfactin forms, and resulted in the appearance of novel surfactin variants with fatty acid chains containing no more than 11 carbon atoms.

Cationic Net Charge and Counter Ion Type as Antimicrobial Activity Determinant Factors of Short Lipopeptides.

To get a better insight into the antimicrobial potency of short cationic lipopeptides, 35 new entities were synthesized using solid phase peptide strategy. All newly obtained lipopeptides were designed to be positively charged from +1 to +4. This was achieved by introducing basic amino acid - lysine - into the lipopeptide structure and had a hydrophobic fatty acid chain attached. Lipopeptides were subjected to microbiological tests using reference strains of Gram-negative bacteria: Escherichia coli, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas aeruginosa, Gram-positive bacteria: Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Enterococcus faecalis, and fungi: Candida albicans, Candida tropicalis, Aspergillus brasiliensis. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimal fungicidal concentration (MFC) were established for each strain. The toxicity toward human cells was determined by hemolysis tests via minimum hemolytic concentration (MHC) determination. The effect of the trifluoroacetic acid (TFA) counter ion on the antimicrobial activity of lipopeptides was also examined by its removing and performing the antimicrobial tests using counter ion-free compounds. The study shows that lipopeptides are more potent against Gram-positive than Gram-negative strains. It was revealed that positive charge equals at least +2 is a necessary condition to observe significant antimicrobial activity, but only when it is balanced with a proper length of hydrophobic fatty acid chain. The hemolytic activity of lipopeptides strongly depends on amino acid composition of the hydrophilic portion of the molecule as well as fatty acid chain length. Compounds endowed with a greater positive charge were more toxic to human erythrocytes. This should be considered during new lipopeptide molecules design. Our studies also revealed the TFA counter ion has no significant effect on the antimicrobial behavior of cationic lipopeptides.

Antifungal Lipopeptides Produced by Bacillus sp. FJAT-14262 Isolated from Rhizosphere Soil of the Medicinal Plant Anoectochilus roxburghii.

This study aimed to develop biocontrol Bacillus and explore bacterial biocontrol substances. According to the blood agar test, strain FJAT-14262 was screened as a biosurfactant-producer. The biosurfactant-producing ability of FJAT-14262 was further confirmed by the oil spreading tests because of its amphipathic character. Furthermore, its fermentation supernatant could decrease the surface tension from 74.1 to 32.7mNm-1. Fourier transform infrared spectroscopy (FT-IR) analysis indicated that the biosurfactant produced by the strain FJAT-14262 was a kind of lipopeptides. Reverse-phase high-performance liquid chromatography (RP-HPLC) and liquid chromatography-mass spectrometry (LC-MS) analysis demonstrated that this lipopeptide contained surfactin with polar amino acids and hydrophobic fatty acid chains. Moreover, bioinformatic analysis revealed that the nonribosomal peptide synthetases genes srfAA, srfAB, and srfAC were structurally conserved in the FJAT-14262 genome. Importantly, the crude surfactant exhibited strong inhibitory activities against Fusarium oxysporum, suggesting that strain FJAT-14262 could be a potential biological control agent against Fusarium wilt.

Kinetics and mechanism of electron transfer reaction of single and double chain surfactant cobalt(III) complex by Fe2+ ions in liposome (dipalmitoylphosphotidylcholine) vesicles: effects of phase transition

In this study, we report the kinetics of reduction reactions of single and double chain surfactant cobalt(III) complexes of octahedral geometry, cis-[Co(en)2(4AMP)(DA)](ClO4)3 and cis-[Co(dmp)2(C12H25NH2)2](ClO4)3 (en = ethylenediamine, dmp = 1,3-diaminopropane, 4AMP = 4-aminopropane, C12H25NH2 = dodecylamine) by Fe2+ ion in dipalmitoylphosphotidylcholine (DPPC) vesicles at different temperatures under pseudo first-order conditions. The kinetics of these reactions is followed by spectrophotometry method. The reactions are found to be second order and the electron transfer is postulated as outer sphere. The remarkable findings in the present investigation are that, below the phase transition temperature of DPPC, the rate decreases with an increase in the concentration of DPPC, while above the phase transition temperature the rate increases with an increase in the concentration of DPPC. The main driving force for this phenomenon is considered to be the intervesicular hydrophobic interaction between vesicles surface and hydrophobic part of the surfactant complexes. Besides, comparing the values of rate constants of these outer-sphere electron transfer reactions in the absence and in the presence of DPPC, the rate constant values in the presence of DPPC are always found to be greater than in the absence of DPPC. This is ascribed to the double hydrophobic fatty acid chain in the DPPC that gives the molecule an overall tubular shape due to the intervesicular hydrophobic interaction between vesicles surface and hydrophobic part of the surfactant complexes more suitable for vesicle aggregation which facilitates lower activation energy, and consequently higher rate is observed in the presence of DPPC. The activation parameters (ΔS# and ΔH#) of the reactions at different temperatures have been calculated which corroborate the kinetics of the reaction.

Para-(benzoyl)-phenylalanine as a potential inhibitor against leptospirosis

AbstractLeptospirosis is a zoonotic disease of global concern caused by Leptospira interrogans. Subtractive genomic approach, metabolic pathway analysis and multi strain genome comparisons of Leptospira interrogans serovars had proposed 88 common drug targets from 5,124 genes of serovar Copenhageni and 4,727 genes of serovar Lai. Three potential drug targets (lpxC, lpxD and lpxB) were identified from Lipid A biosynthesis process of lipopolysaccharide (LPS) biosynthesis pathway. Lipid A is one of the three components of LPS that contains multiple hydrophobic fatty acid chains which anchor the LPS into the bacterial membrane. Designing inhibitory drug molecules targeting Lipid A biosynthesis would dissolve the structural integrity of membrane structure leading to cell lysis and death of Leptospira. LpxC being the first enzyme among the three drug targets of Lipid A biosynthesis pathway; blocking the enzyme with suitable inhibitor would stop synthesizing substrates for lpxD and lpxB. Also, there is no alternative mechanism in Leptospira to replace the catalytic activity rendered by lpxC; hence, the drug target was selected herein for rational drug design. The lpxC tertiary structure was modeled incorporating inhibitor BB-78485 using Modeller9v8. The lpxC 3D structure reliability was assessed through various model validation techniques. Ligand based virtual screening was performed from one million entries of ligand.Info metadatabase. The leads were ranked using computational docking technique of Glidev5.5. Twelve leads with better binding affinity than BB-78485 (XPGscore -7.98 Kcal/mol) were proposed as potential inhibitors of lpxC. Para-(benzoyl)-phenylalanine, the best ranked inhibitor (XPGscore -10.34Kcal/mol), would be intriguing for rational drug design against leptospirosis.

Nonhydrolyzable analogs of phosphatidylinositol as ligands of phospholipases C

Phosphatidylinositol-specific phospholipases C (PI-PLCs) are important enzymes participating in transmembrane signal transduction. The structures of the two major species of these enzymes: bacterial Ca2+-nondependent enzyme from B. cereus and mammalian Ca2+-dependent PLCδ1 from rat brain in the complexes with the polar head groups of their substrates have been previously solved. Although these structures show few differences as compared to free enzymes, there is a compelling evidence that full catalytic activity of PI-PLC necessitates interaction of the enzyme with the entire substrate, including the hydrophobic fatty acid chains. In this work we develop new tightly binding and cleavage-resistant analogs of phosphatidylinositol, using relatively minor modifications of the structure. Two synthesized analogs, 2-amino-2-deoxy-PI (8) and the conformationally constrained analog (10) had binding affinities (Ki) in 10 μM range. 15N-1H HSQC NMR spectra of uniformly 15N-labeled bacterial Ca2+-nondependent and Ca2+-dependent phospholipases C, btPLC and saPLC1, respectively, displayed changes upon ligand binding that suggest an occurrence of a conformational change.

Liposome mediated modulation of macrophage functions.

Liposomes are artificially prepared spheres, consisting of concentric phospholipid bilayers separated by aqueous compartments. They form, when phospholipids e.g. phosphatidylcholine molecules, are dispersed in water. The phospholipid molecules will find a conformation in which their hydrophobic fatty acid chains are prevented from contact with water. For that reason, phospholipid bilayers are formed in which the relatively hydrophilic head groups are making up both of the outer parts of each bilayer, whereas the hydrophobic fatty acid groups are located directly opposed to each other in the inner side of the bilayer. Part of the aqueous solution together with hydrophilic molecules, solved in it, will be encapsulated during the formation of the liposomes. Lipophilic molecules will be associated with the phospholipid bilayers themselves. The hydrophobic parts of amphipathic molecules will be inserted in the bilayers, whereas their hydrophilic parts are extending in the aqueous compartments or are exposed on the outer surfaces of the liposomes (figure 1). Numbers of concentric phospholipid bilayers (unilamellar and multilamellar liposomes), phospholipid composition and charge of the liposomes can be varied. Targeting of the liposomes may be achieved by the insertion of target molecules (e.g. monoclonal antibodies or sugar residues) in their outer (surface) bilayer (Gregoriadis, I988).

Cyclodextrins lessen the membrane damaging effect of nonionic tensides

Nonylphenyl-ethyleneoxide polymers containing 5, 9 and 30 ethyleneoxide groups per molecule build into the hydrophobic fatty acid chains of the cell membrane phospholipid dipalmitoyl-phosphatidylcholine (DPPC) resulting in a decreased main transition temperature, a decreased enthalpy of the main transition and in enhanced potassium permeability of DPPC liposomes. The α-, β- and γ-cyclodextrins form inclusion complexes with the tenzides lowering their free concentration. The complex formation lessens or sometimes totally prevents the membrane damaging effect of tensides. The effectivity order of cyclodextrins is βCD>γCD>αCD.

The phospholipid requirement for RH 0(D) antigen activity: Mode of inactivation by phospholipases and of protection by anti-Rh 0(D) antibody

Previous studies have suggested a membrane phospholipid requirement for Rh 0(D) antigen activity. Isolated erythrocyte membranes incubated with phospholipase A 2 from both bee venom and porcine pancreas undergo loss of Rh antigen activity. The mode of attenuation of this antigen activity as indicated by double-reciprocal binding plots suggests substantial loss of sites accompanied by an apparently increased association constant. In the presence of anti-Rh 0(D), but not anti-A, bound to group A Rh 0(D)-positive membranes prior to hydrolysis, there is marked protection: almost complete preservation of sites at the expense of a decreased association constant. This pattern of protection is not seen with phospholipase C, which cleaves the polar headgroup in contrast to the A 2-enzymes, which hydrolyze the fatty acid in the 2-position. Analysis of the products of digestion shows a trend to protection of bulk phospholipids of all major classes in the presence of bound specific antibody. The hydrophobic fatty acid chain may be the site with which the bound anti-Rh 0(D) antibody is in closest proximity.

13C nuclear magnetic resonance spectroscopic evidence for hydrophobic lipid-protein interactions in human high density lipoproteins.

Phosphatidylcholines, sphingomyelins, cholesterol, and cholesterol esters were enriched with (13)C by chemical synthesis in specific positions of their hydrophilic groups and aliphatic chains. Their spin-lattice relaxation times were determined in organic solvents. The substances were organized as liposomes and recombined with total human high density apolipoproteins and the two separated main components, apolipoprotein A-I (apoLp-Gln-I) and apolipoprotein A-II (apoLp-Gln-II). These (13)C nuclear magnetic resonance data established that in reassembled high density lipoproteins the phospholipid molecules bind to the apoprotein moieties with their hydrophobic fatty acid chains and not with their hydrophilic zwitterionic groups. Apolipoprotein A-I preferentially binds phosphatidylcholine, although its lipid-binding capacity is smaller than that of apolipoprotein A-II. Apolipoprotein A-II avidly reassembles with sphingomyelin by hydrophobic interactions. A model of the molecular organization of the high density lipoportein particle has been derived.

Reversal by phospholipids of the oligomycin induced inhibition of membrane associated adenosintriphosphatases

The inhibitory effect of oligomycin on sodium, potassium-stimulated ATPase and on mitochondrial ATPase was removed by the addition of purified phospholipids to the incubation medium. Ouabain effect was not modified. Phospholipids with isoelectric ionic portion (particularly lecithins) were found more active than phosphatidylethanolamines and bovine brain phosphatidylserine. Differences in the activity of lecithins from various sources were also seen. This indicates that the composition of hydrophobic fatty acid chains influences the reversal of oligomycin effect. The antagonism by the external added phospholipids may reflect the affinity of oligomycin with the phospholipids of the enzyme preparation.

The displacement of calcium ions from phospholipid monolayers by pharmacologically active and other organic bases.

1. The binding of (45)Ca(2+) to a monolayer of phosphatidylinositol at the air-water interface was maximal when the separation of the phospholipid head groups approximated to the diameter of a hydrated Ca(2+) ion. 2. The displacement of Ca(2+) adsorbed on monomolecular films of phosphatidylinositol by a series of drugs (both narcotic and excitatory) and other organic bases was related to the ability of the bases to penetrate into the film. 3. With films of phosphatidylinositol at constant area, and at an initial surface pressure of 10dynes/cm., the displacement of Ca(2+) by increasing concentrations of the local anaesthetic, tetracaine, was linearly related to the change in surface pressure (Deltapi) caused by the penetration of the drug. 4. Deltapi and the displacement of Ca(2+) showed a related fall when the initial surface pressure of the phosphatidylinositol film was increased from 4 to 40dynes/cm. both at a constant bulk tetracaine concentration and when this latter concentration was adjusted to keep it at a constant ratio to the surface density of phosphatidylinositol molecules. 5. The displacement of Ca(2+) from phosphatidylinositol films by cetyltri-methylammonium ions was directly compared with the surface concentration of the base in the film, measured by using labelled base and a surface-radioactivity technique. 6. The ability of a series of straight-chain aliphatic amines to displace Ca(2+) from phosphatidylinositol films increased with the number of carbon atoms up to C(12). However, there was a marked jump in the displacing activity after hexylamine, and this could probably be correlated with the carbon chain's being of sufficient length to just reach the hydrophobic fatty acid chains of the orientated phospholipid molecules with the charges on both substances in juxtaposition.