Current Opinion in ea ELSEVIER Current Opinion in Cold & lnrfae Scene 2001) 144-159 Natural surfactants Krister Holmberg* erent of pli Sue Chemisty, Chane: Unie of Teche, SE-12 9 Gite Sen Apstract “The ever-increasing environmental concern about surfactants triggers an interest in natural surfactant. This review, which ‘hasan emphasis on work published since 1908, covers three categories of natural surfactants: amphiphiles produced by yeast or bacteria, amphiphiles containing a natural polar headgroup and amphiphiles containing a natural hydrophobic tai “Microorganisms produce both high molecular weight and low molecular weight surfactants. Only the low molecular weight ‘compounds are included in the review. Sugars and amino acids are the two most important examples of surfactant polar Fheadgroups of natural origin. The research is particularly intense in the arca of sugar surfactants and the review covers three types: allyglucosides, akyglucamides and sugar esters. Surfactants based on two types of natural hydrophobic tails are inchided: fatty seid monosthanolamides and sterol ethorglates, Routos of preparation ax well ar physico-chemical proportion are discussed forthe surfactants prepared by organic synthesis. © 2001 Elsevier Science Ltd. Al rights reserved, Keywords: Surfactant; Naural Biotechnology: Sugars Pll: Amino ac; Fat aids Steal 1. Introduetion ‘The term ‘natural surfactant’ iS not unambiguous Taken strictly a natural surfactant is a surfactant taken directly from a natural source, The source may be of either plant or animal origin and the product should be obtained by some kind of separation proce- dure such as extraction, precipitation or distillation. No organic synthesis should be involved, not even as fan alter-reatment, There are in fact not many surfac- tants in use today that fulfil these requirements, Lecithin, obtained either from soybean of from egg yolk, is probably the best example of a teuly natural surfactant. "The main reason why natural surfactants in the real sense of the word are so scarce is not a lack of availability. Amphiphiles are abundant in both the Tek: $46.31-7722965 fae 44631-16062 E-mail ads: Kh@surchem chalets se (K. Holbers plant and the animal kingdom where they are often referred to as polar lipids. In biological systems the surface active agents are used in very much the same way as surfactants are employed in technical systems: to overcome solubility problems, as emulsifiers, as dispersants, to modify surfaces, etc. The factor that works against production of truly natural surfactants is the cost of work-up. The products are usually present in small quantities and the separation process tends to be tedious. In most instances the cost of separation /solation will by far ‘exceed the manufacturing cost of equivalent synthetic surfactants. The unfavorable cost situation for truly natural surfactants may change if fermentation processes can be developed that yield biosurfactants in high yields. Both yeast and bacteria can be efficent producers of surface active agents and there is considerable cur- rent interest in biotechnological processes for produc- tion of amphiphilic compounds. These substances can be either low molecular weight, such as acylated 13590294 /01/$ se font matter 2001 Ehevier Seienoe Li. All rights served, Pu $1359°0294(01)00074-7K. Holmbrg / Curent Opinion Colo & Interface Since 6 2000) 148-159 9 oligosaccharides, or high molecular weight, most of- ten lipopolysaccharides. So far most interest has fo- cused on the high molecular weight products, with © Emulsan, a highly charged heterolipopolysaecharide with a molecular weight of approximately 10°, as the prime example [1]. ‘The term ‘natural surfactant’ is often used in a broader sense than that discussed above, however. Surfactants synthesized from natural raw materials are usually referred to as natural surfactants, Proper ‘examples of surfactants belonging to this category are tally acid esters of sugars and Tally acid esters oF amides of amino acids. Its, in fact, common practice to be even mote generous in the use of the term “natural surfactant’, A surfactant with one of the main building blocks, the polar headgroup or the hy- drophobic tail, obtained from a natural source is often referred to as a natural surfactant. For example, alkyl glucosides which are prepared from a ‘natural’ sugar unit and a ‘non-natural’ fatty aleohol are often regarded as natural surfactants. (So-called ‘natural fatty alcohols’ should not be seen as natural starting ‘materials, although this is often done. Fatty alcohols derived from coconut oil and other triglycerides are made by two consecutive chemical reactions: rmethanolysis of the triglyceride followed by reduction of the methyl ester.) In this review the term “natural surfactant’ is used in its broadest sense. The paper covers true natural surfactants prepared by fermentation, synthetic sur- factants with a natural polar head group and synthetic surfactants with @ natural hydrophobic tail. Only low molecular weight surfactants with a well-defined structure are included, The ‘natural surfactants’ used to treat the respiratory distress syndrome, a chapter of its own, are in reality mixtures of amphiphiles with structures not fully elucidated (2] and are, hence, not covered. The review is organized correspondin, 2. Surfactants prepared by fermentation 21. Agtpolvols Acylpolyols prepared by fermentation are usually (Hidy~Gi-d ge (CHa -CO-CH-CHOH-(CH,)q-CHy ge O—CO-CH-CHOH-(CH;)q-CHy, 1 ie hydroxy faty acids connected to disaccharides by es- ter bonds. ‘They are extracellular compounds pro- duced by. actinomyeetes such as Mycobacterium, Corynebacterium and Brevibacterum (3). The acylpo ‘ls are abundant in bacterial cell wall. A typical example of an aeylpolyol isthe trehalose ester shown in Fig 1. 2.2. Glcolipids Giycolipids are usually hydroxy faty acids attached to a sugar via a glycosidic bond. Sophorolipids and ‘hamnolipids are well known examples, produced by Candida and by Pseudomonas, respectively [4,5]. Fig. 2 shows a cyclic and an acyclic sophorolipid. Rham- nolipids are attractive in that they can be efficiently produced during growth on either hydrocarbon of carbohydrates as the sole carbon source. There has been considerable interest in the molecular genet ‘of rhamnolipids and this topic has recently been re- viewed in this journal (6). ‘A sucrose lipid produced by Serratia marcescens has recently been isolated and characterized (7*]. ‘The product was found to be an excellent emubifier for a sae HOA Ae eo O : LN f OH oe iw on Fig.2. A gclic and an acetic sophoroipi150 K Holmberg / Carte Opinion in Coli & nerve Scene 6 (201) 148-159 ct exotic {Gly —U-Le Lev —L-ValL-Asp—Dstes bebe ° Fig. Surctin om Bucs subi Wide range of hydrocarbons including crude oil. 1 has been suggested as a surfactant for cleaning of oil tanks. 23. Aoslpepiides Acylpeptides or lipopeptides, are usually cyclic compounds based on a hydroxy acid and a short peptide chain, By far the most studied compound is *Surfactin, produced from Bacillus subulis [8:9 Acylpeptides, such as Surfactin and Lichenysin_pro- duced by Bacillus licheniformis, are efficient am- phiphiles when it comes to surface tension reduction, tc, Surfactin is one of the few biosurfactants that has found commercial use. It is used in a variety of pharmacological applications [10]. The formula of Surfactin is shown in Fig. 3. Susfactin and rham- rolipids are the only two biosurfactants for which there is any regulatory information and the molecular genetics of Surfactin has recently been reviewed [6] chow ° ide gc ae a a Di gcse | : Glucamide oon on 4 PESO a Ty Fig. 4. Examples of pobol surfactants. From top (© bottom: an ally lucoside, an alglcamie, and the ety! acetal of ty ic vom eter 3. Surfactants based on a natural polar headgroup Sul. Sugar as polar headgroup Various types af sugars or polyols derived from sugar have been used as surfactant polar headgroup for many years. Sorbitane alkanoates and ethoxylated sorbitane alkanoates, well known under the trade names of ® Span and ® Tween, respectively, have been around for along time. In recent years there has been ‘a focus on three classes of surfactants with sugar or a polyol derived from sugar as polar headgroup: alkyl polyglucosides (APGs), alkyl glucamides and sugar esters. Representative structures of the three surfac- tant types are shown in Fig. 4 ‘There is currently a very strong interest in ex- ploring alkyl polyglucosides (APGs) as surfactant for several types of applications. APGs are synthesized by direct reaction of glucose with fatty alcohol, using a large excess of alcohol in order to minimize sugar oligomerization. Alternatively, they are made by transacetalization of a short chain alkyl glucoside, such as ethyl or butyl glucoside, with a long chain alcohol. An acid catalyst is used in both processes. Either glucose or a degraded starch fraction is used as starting material, Fig. 5 illustrates the synthesis. Alkyl ns a veo mame ey ‘ ta eon, ) | wea ae Reena Fig 5. Pathway for APG sates liom von Rybiaski and Hi [18) with permissionK. Holmbrg / Curent Opinion Colo & Interface Since 6 2000) 148-159 as slucosides can also be made by enzymatic synthesis, using B-glucosidase as catalyst which yields only the Branomer (and gives low yield). The corresponding eanomer can more really be obtained by Prgluco- sidase catalyzed hydrolysis of the racemate, There are considerable differences between the af mixture ob- tained by organic synthesis and the pure enantiomers obtained by the bio-organic route. The B-anomer of rn-oetyl glucoside has found use as a surfactant in biochemical work. Alkyl ghucosides are stable at high pH and sensitive to tow pH where they hydrolyze to sugar and tatty aleohol. A sugar unit is more water-soluble and less soluble in hydrocarbons than the corresponding poly- oxyethylene unit; hence, APGs and other poiyol-based surfactants are-more lipophobie than their poly- onyethylene-based surfactant counterparts [11]. This ‘makes the physico-chemical behavior of APG surfac- tants in oil-water systems distinctly different from that of conventional non-ionics. Furthermore, APGs do not show the pronounced inverse solubility vs temperature relationship that normal non-ionies do. This makes an important difference in solution behav- jor between APGs and polyoxyethylene-based surfac- tants, The main attractiveness of APGs lies in their favorable environmental profile: the rate of biodegra- dation is usually high and the aquatic toxicity is low [12,13] In addition, APGs exhibit favorable dermato- logical properties, being very mild to skin and eye {14}. ‘The mildness makes this surfactant class attrac- tive for personal care products but APGs have also found a range of technical applications. APGs have been the topic of several excellent overviews [15-18]. In addition, the physico-chemical properties of APGs, as well as other polyhydroxy based surfactants, have recently been reviewed in this journal [19]. Only a few brief notes on recent develop- ‘ments in the field will therefore be given here ‘The solution behavior of APGs, both anomeric mixtures, prepared by organic synthesis, and pure enantiomers have been studied in detail (20°,21-23} The micellar phase region is usually large ‘and at higher concentrations the normal pattern of liquid crystalline phases appears. It interesting to note, as was described in a previvus review [19} dat some ‘APG surfactants exhibit & miscibility gap in the mice! Jar region. As with polyoxyethylene-based non-ionies two micellar phases appear in this region, one dilute and one concentrated. For a pure decyl glucoside surfactant it was found thatthe dilute phase consisted of micelles of aggregation number in the range 200-400 while the concentrated phase contained larger aggregates, probably branched micelles that formed a network through entanglement [20]. A ehar- acteristic feature of the liguid exystalline region which appears at higher surfactant concentration is that the borders between the different crystalline phases in a temperature vs. surfactant concentration diagram are almost vertical, indicating a temperature-independent ‘behavior. This is very different from the behavior of polyoxyethylene-based non-ionics. Mixed micelle formation of two APG with differ- cent length of the alkyl chain with both anionic and cationic surfactants have been studied [24°]. The ex- perimental data were fitted to Rubingh’s model for rnon-ideal mixed micelle formation [25], value of the interaction parameter, B is obtained and the value of B, positive or negative, indicates whether there is a repukive or an attractive interaction involved in the formation of mixed micelles between the two surfac: tant types. Combination of an APG with the cationic surfactant dodecyltrimethylammonium bromide gave a value of the B-parameter of —4.1 indicating rather strong attractive interaction. The combination of the same APG with the anionic surfactant sodium dode- eylsulfate gave a B-value of —23, indicative of a smaller attraction. These results are interesting and not easily explained. They tend to support the view, proposed by several workers, that APGS carry a nega” tive net charge, the origin of which is unclear [24°]. It is interesting that whereas a combination of an APG and an alcohol ethoxylate gave a value of the B- parameter of zero, the same APG when combined with another alkyl plycoside, an alkyl maltoside of the same alkyl chain length (C10) gave a -value of ~ 2.0, This attractive interaction was tentatively explained as due to favorable packing of the headgroups of the 1wo surfactants [24° ‘The majority of work on glycoside surfactants has been made on alkyl glucosides, ie. surfactants based ‘on glucose or slightly oligomerized glucose as polar headgroup. Recently, alkyl maltosides have been stud- ied with a specific focus on its behavior in. mixed micelles together with anionic and non-ionic surfae- tants [26,27]. A combination of a decyl maltoside with sodium dodecylbenzene sulfonate gave a value of the B-parameter of ~2.1 which ean be compared with the value of ~3.3 for a combination of an alcohol ethoxy- late, octa(ethylene glycol)monodecyl ether (C10E8) with the same anionic surfactant. combination of Ue alkyl mvaltoside with de acolo! elhylave gave at very small B-value (—03). A synergistic effect with regard to surface tension reduction was also found for combinations of cither of the non-ionic surfactants and sodium dodecylbenzene sulfonate. Thus, the re- sults obtained with the maltoside surfactant agree very well with those previously obtained with gluco- side surfactants of the same alkyl chain length. A full explanation of the results still remains, however. Alkylglucamides (see Fig. 4), or more strictly named as Nealkanoyl-N-metylglucamines, are commercially important products. The product sold in large quant192 K Holmberg / Carte Opinion in Coli & nerve Scene 6 (201) 148-159 ties for the detergent sector is N-dodecanoyl-N-meth- vlglucamines, ic. the C12-derivative, The product prepared from glucose, metyl amine, hydrogen and ‘methyl laurate by a two-step reaction ‘Alkylglucamides have been the subject of an exten- sive review recently [28]. Few major investigations on this surfactant class seem to have been made recently. In a paper by Zhu et al, a gluicamide was compared with polyol surfactants of similar structure, a xylamide and a glyceramide, all having a C12 alkyl chain [29°] ‘The glucamide has four hydroxyl groups in the polar head, the xylamide three and the glyeeramide one. ‘This was reflected both in the area per molecule values and the CMC values which were 30, 28 and 26 ¥ and 0347, 0.331 and 0.234 mM, respectively. here is considerable current interest in sugar es- tors (see Fig. 4) and a recent review covers the topic until 1998 [30]. Esters of glucose can be made either by enzymatic synthesis, using a lipase catalyst, or by ‘an orgunic chemical route. Using the right enzyme the bio-organic route can give esterification almost exclusively at the 6-position of the sugar moiety. The ‘organic synthesis requires extensive use of protecting groups to obtain high selectivity, Selective enzymatic synthesis of other sugar esters than glucose esters hhave been difficult t0 achieve without the use of protective groups. Starting from sugar acetals and fatty acids. monoesters of sugars were obtained in good yield, after a deprotection step, from several Starting materials, both mono: and disaccharides (132) ‘The advantage of the organic synthesis over the enzymatic route is that the sugar unit cam be varied at will, The group of Drummond has recently made important contributions in this area with a specific focus on degradation mechanisms [33 5] but also covering physico-chemical behavior [36*]. A se- ties of sugar ester surfactants were synthesized having different headgroup size (glucose, sucrose and rafti- nose, consisting of one, wo and three anhydroglucose rings, respectively) and with C12 and C16 acyl chains. Sulfonated sugar esters were also prepared. It was found that all unsubstituted sugar esters degraded rapidly. Variations in the sugar headgroup size or the acyl group clint Feugtl did ot siguificantly affect Ue biodegradability. The anionic derivatives degraded more slowly, however [33] ‘A detailed study on the mechanism of biodegrada- tion of the unmodified and the sulfonated sugar esters showed that sucrose lnurate biodegradation occurs vi initial ester hydrolysis. The sucrose a-sulfonyl laurate, fon the other hand, degrades by initial alkyl chain oxidation, This indicates that the ester hydrolysis pathway is blocked by the sulfonyl group adjacent to the ester bond so that biodegradation is forced 10 proceed via the slower alkyl chain oxidation pathway [34°*], An ethyl substituent in a-position instead of the sulfonate group also slowed down the biodegrada- tion rate, probably for the same reason, The two degradation routes are shown in Fig. 6, Base catalyzed hydrolysis. was also investigated. Similar to the results from the biodegradation, the unsubstituted sugar esters degraded faster than the derivatives with either a sulfonate or an ethyl group in the a-position, The difference in hydrolysis rate between the three surfactants suerose laurate, sucrose ‘sulfonyl laurate and suerose a-ethyl laurate, varied with temperature, It was concluded that at’ higher temperature, when steric hindrance is the dominant factor, hydrolysis of a-sulfonyl laurate is slowest. By contrast, at lower temperature, when activation en- ergy is more important, hydrolysis is faster in the presence of an electronswithdrawing a-sulfonyl group than in the presence of an electron-attracting a-ethyl group [35], Dimeric and trimeric sugar ester surfactants have been synthesized by a combination of enzymatic and ‘organic chemical methods [37%], An impressive se- ries of products were prepared with different spacer units between the sugar tings. For instance, a dimeric (gemini) glucose ester with C12 acyl chains attached at positions 6 of the two anhydroglucose units and with the two moieties connected via a six carbon spacer was prepared in high yield. The product is shown in Fig 7 3.2. Amino acid as polar headgroup Amino acids and short peptides constitute an alter- native to sugars as a natural polar headgroup for surfactants, Several amino acids have been investi- gated for this purpose with the majority of papers dealing with basic amino acids such as arginine and Iysine from which cationic surfactants can easily be prepared, Most synthetic procedures ate based on ‘organic synthesis but enzymatic processes have also bbeen explored [38,39]. Arginine-based surfactants have recently been reviewed [40 Arginine surfactants, such as N-dodecanoylarginine ‘metyl ester salt are conveniently prepared by reaction between the vil soluble doducanoic acid and the water soluble arginine methyl ester hydrochloride using di cyclohexylearbodiimide as condensing agent. The re- action is normally carried out in an aprotic, polar solvent such as dimethylformamide (DMF). The reac- tion is performed at room temperature and works: well in a laboratory scale. However, DMF is an un- suitable solvent for large scale operations, partly due to cost and toxicity and partly due to difficulties in removing it from the product by simple vacuum evap- oration. An alternative process, using a high internal-phase-ratio emulsion, has been investigatedK. Holmbrg / Curent Opinion Colo & Interface Since 6 2000) 148-159 153 Be ‘eg? (pester he howe ci Sone (och, Ae A 9 e081 2) woxidation Soew, 0 Hoey, ‘ot 19 10) 0% \Veu,on Wolf Fig. 6 Degradation pathways of «sugar fatty acid ester (rom Baker etal [4] with permsion) for the synthesis [41°], Such emulsions can be formu- lated with a large amount, sometimes above 99%, of, internal phase and with very low amount of surfac- tant, often less than 0.5%. The structure of water-in- oil Emulsions of this type is that of closed-packed Q OCtCH Cy S70, tall Ho: on A 0. aN ‘on oH cHyichiego © Fig. 7. A sugar fay aid ge wih permission surfactant om Gao eta. (37) water droplets of a broad size distribution separated by a thin film of the continuous oil phase. The vis- cosity is usually high and such emulsions are sometimes referred to as gel emulsions. It was de- ‘onstrated that the concentrated emulsions were good reaction media with yields equivalent to those ‘obtained in DMF. The yield depended on the water- to-0il ratio. Too high water content at constant sur- factant concentration gave large water droplets with corresponding small interfacial area which was found to be unfavorable. A papain-catalyzed synthesis of arginine-based sur- factants have been reported [42,43]. Papain deposited ‘on a solid support was used as catalyst for the amide and ester bond formation between arginine methyl ester and various long-chain alkyl amines and fatty6 K Holmberg / Carte Opinion in Coli & nerve Scene 6 (201) 148-159 Tohene * HOR -CH-CHyCl+ ROH —_H:S0, ti eee Clon bc ‘goon giron c,d, . won + Abii MeOH 4 NOH oa-c1 a } “He WW éwon HOR eI Fig.§. Rowe of preparation of typtophanthased surf alcohols in organic solvent. High yield of arginine amide was obtained from reaction in acetonitrile. For the synthesis of the arginine fatty acid ester the best yield was obtained in a solvent-free system kept at a temperature above the melting temperature of the fatty alcohol Using a combination of organic and bioorganic synthesis arginine-based gemini surfactants have been synthesized by the group of Infante [44] and the solution properties have been investigated [45], The gemini surfactants consisted of two N-acylarginine ‘moieties connected via the arginine carbonyl groups by @ diamino spacer molecule, such as 13-4 aminopropane or 1,3-diamino-2-hydroxypropane. A Neacyl-Larginine alkyl ester was used as. starting material, This compound was first reacted with the ddiamino spacer to form the amino-amide, ie. the spacer reacted with one of its amino group to form a N-aeyl-t-arginine amide. In a second step, catalyzed by papain, the free amino group reacted with another molecule of N-acyl-targinine alkyl ester to form the diamide product. Overall yields of the order of 65% were reported. precipiation stant (fom Pegiadvo et al [45] with permision, A series of tryptophan-based surfactants have been synthesized [46°]. The surfactants were synthesized by reaction of yptophan with the epichlorohydrine adduet of a fatty alcohol, as shown in Fig. 8. The series of surfactants based on fatty aleohols with from 9 to 16 carbon atoms was evaluated with respect 0 ‘CMC, surface tension at CMC and area per molecule at the air-water interface. The results are summa- rized in Table 1. As can be seen, the CMC values are low, considerably lower than for simple alkanoate surfactants of the same hydrocarbon chain length. For instance, sodium laurate has a CMC of 20 mM to be compared with the value of 0038 mM of the C12 surfactant of Table 1. The surface tension values are generally low, as expected for these hydrophobic sur- factants. The areas per molecule are within the ex- pected range, the increase in area for the shorter chain length surfactants: most probably reflecting @ less ordered packing at the surface. ‘The amino acid-based surfactants are possible can- didates for pharmaceutical applications and the hemolytic action of some Iysine-based anionic surfac- tants has been investigated. The surfactants were saltsK. Holmbrg / Curent Opinion Colo & Interface Since 6 2000) 148-159 Table 1 Prysc-chemical data fr arinie-baed surictants having the structure given in Fig 8 (data 155 kon from Pegiadu etal. [460 Compound eMC ‘Serfae tension at CMC ‘Area por molecule Gann, (N/m) a R-onI 05 = R= CIOHL 136 o R—CuDS is S R=cIIDS ms 51 RCI nz 6 Racin ‘nm 6 ROCISHL nr rs of N,.N.-dialkanoyl lysine. In one series of experi- ments the alkyl chain lengths were vatied [47]. In another work the surfactant was N, .N,-octanoyl ly- sine and the counterion was varied [48]. As expected, the hemolytic activity depended on the alkyl chain Tength and on the choice of counterion, With Iysine as counterion to the lysine-based anionic surfactant a protective effect against hypotonic hemolysis was ob- tained. 4. Surfactants based on a natural hydrophobic tail “4d. Fatty acid as hydrophobic tail Fatty amide cthoxylates are easily prepared by cthoxylation of the fatty amide monoethanolamide. ‘The ethanolamide, in turn, is prepared by aminolysis, of the fatty acid metyl ester by ethanolamine. The fatly amide ethoxylates are of interest as an alterna- tive to faty alcohol ethoxylates for several reasons: () they biodegrade readily to fatty acid and amino- terminated poly(ethylene glyco) (i) the amide bond in the structure may improve surfactant packing due to hydrogen bond formation; and (ii) double bonds in the fatty acid chain can be preserved in the product (although during the ethoxylation step 1,3-ciscs-dou- ble bonds undergo migration to conjugated ‘rans.cis structures), A series of fatty amid cthoxylates, all based on C18. fatty acids but with varying number of double buns, hhas been synthesized and evaluated with regard to the effect of the amide bond and the double bonds on physico-chemical properties [49*.50*]. Surfactants ‘with different polyoxyethylene chain lengths were pre- pared and the corresponding saturated fatty alcohol ethoxylates, stearyl ethoxylates, were used as refer- ences. It was found that the presence of the amide group led to a decrease in CMC, possibly due to hydrogen bond formation. The surface tension at the CMC was higher when an amide bond was present, however, which was assumed to reflect the increase in size of the polar group (decrease in the critical pack- ing parameter value) which is unfavorable for close alignment of surfactant molecules at the air-water interface. The same effect was seen in adsoeption at a hydrophobic, solid surface, as studied by ellipsometry. The presence of unsaturations in the hydrophobic tail gave an increase in CMC. This may partly be due to the surfactants becoming more hydrophilic and partly to the increased bulkiness of the chains render- ing packing into closed aggregates and formation of hydrogen bonds between amide groups more dificult. ‘The presence of one double bond, cis (oleyl) or trans (claidyD, did not much affeet the molecular cross-see- tional area at the surface, Two double bonds gave ‘nivel larger aveas, as ean be seen fiom Fig. 9. Self-diffusion NMR was used to study the effect of temperature and surfactant concentration on the size of the surfactant aggregates. It was found that at both Cross Sectional Headgroup Area / A? Number of Oxyethyiene Units Fig. 9. Area per molecule for series of flty amie etholtes ith 1015 or 20 onetilene unis The number of double bonds 8 {he acl chain vay from zero (seat to one, Coes and on, ‘rons (eids) oto CinoleyD, Stary alah erhssates (CISEn) fre incded a references.150 K Holmberg / Carte Opinion in Coli & nerve Scene 6 (201) 148-159 high and low surfactant concentration the hydrody- namic radius of a stearyl amide ethoxylate was some- what larger than the radius of the stearyl alcohol ethoxylate with the same number of oxvethylene units. For the higher surfactant concentration, where the ‘micelles are non-spherical, this may be due to the larger polar headgroup size of the amide surfactants (ee discussion above) giving slightly less elongated micelles. The results are more difficult to rationalize for the low surfactant concentration where both am- Dhiphiles give spherical micelles. 42, Sterol as hydrophobic tail Sterol-based surfactants have been reviewed re~ cently [SI] and a new review is in progress (52) Sterol-based surfactants are of interest because of the large hydrophobic group of fully natural origin which, due to its rather planar four ring structure, may induce good packing at interfaces. Two recent papers deal with phytosterol ethoxylates {53%,54]. Phytos- terols are sterols of plant origin and their structure is Similar to that of cholesterol, the prime example of sterols from animal sources. The sterols contain a secondary hydroxyl group that can be ethoxylated. ‘The alcohol is sterically hindered, however, and the reaction with ethylene oxide is not straightforward. ‘The host procedure seems to he ta start the ethoxyla- tion with a Lewis acid catalyst, cease the reaction after 3-5 mol of ethylene oxide have been added, and continue with KOH as initiator [55]. The structure of a representative sterol ethoxylate is shown in Fig. 10. Surface tension plots of a series of phytosterol cthoxylates with 10, 20 and 30 oxyethylene units showed that (i) the CMC values were very low; and Gi) the CMC increased with increasing polyoxyethy- lene chain length. ‘The latter, which has previously been observed by others as accounted for in Folmer ct al, 53°], is unexpected! and is difficult to explain. ‘The values are collected in Table 2. Another interest- ing observation is that the time to reach surface tension equilibrium is very long, more than 2 h. The decay of surface tension with time is shown in Fig. 11. ‘Most likely, the long equilibrium time is due t0 ex- HO(CaH,0}¢7 Fig. 10, Sitosteol eons Table? Pyscochemical data for sero-based surfactants having the structure given in Fig 10 ta taken from Folmce eta [53D Nanberot one Sure opetinene Gap tension at CMC CaN md 0 0 u » 7 ca hunge reactions of sterol ethoxylawes with varying polyoxyethylene chain length at the air-water inter- face and to slow conformational changes of individual ‘molecules at the surface. The multiring structure of sterols is rigid and the time required to adapt a favorable conformation at an interface may be long. ‘The phase behavior of the sterol ethoxylates. has been studied in some detail [53°,54], Fig. 12 shows phase diagrams for phytosterols with 5, 10, 20 and 30 ‘oxyethylene units, The surfactant having 10 oxyethy lene units gives a lamellar phase over a very wide concentration range at elevated temperature. The phase diagrams of the more hydrophilic surfactants With large headgroups are dominated by hexagonal ‘and cubie phases. The high thermal stability of the liquid crystalline phases is noteworthy. It is particu- larly striking to see a cubic phase for a non-ionic surfactant that extends above 100°C. Cubic phases are ‘normally seen as being trapped in a three-dimen- sional lattice with no possibility of movement. Usually when the temperature is increased a cubie structure therefore melts at a relatively low temperature. The high thermal stability of the cubie phase formed by the ethoxylated sterols is probably related to an effi- cient packing of the sterol moieties. Self-diffusion NMR measurements of samples trom within the cubic 65 Surface Tension / mN m? 0 2,000 4,000 6,000 8,000 ‘Time /s Fig. 1, Surfice tension for phytstro with 2) caethyene units ‘8 funtion of ime at fixed eonesniation (3.82 10" MY Crom Folmer etal (5) with permission).K. Holmbrg / Curent Opinion Colo & Interface Since 6 2000) 148-159 i one “Temperate °C eae oe c o - 2 £ u/n} nr \t2| i i i i i i aa ew Surfactant wt Fig 12. Phase digeams for phytsteral with (a) Sve oxetylene ‘mts 6) 1 exgttylne units (e) 20 oyetylene uss and) 30 following phases: micela, reversed micellar, lamellar, gel lose picked micellar cubic, nomati, and hexagonal espectively (roms Folmer ea. (33 wth permission) phase has shown the presence of elongated aggre- gates with an axial ratio of 1.6 [54] Conclusions ‘The review has covered work performed in recent years in three areas, ie. surfactants produced from bacteria and yeast, surfactants based On either sugar fof amino acid as polar headgroup, and surfactants based on cither fatty acid or sterol as hydrophobic tai, All three areas are important and subject to considerable research activity. The natural surfactants obtained by these approaches are generally well char- acterized and the performance are up to the stan- dards of conventional surfactants. The biotechnologi- cally produced surfactants suffer from high cost due to relatively low yield in the fermentation and cum- bersome work-up procedures. The surfactants based oon either a natural polar headgroup or a natural hydrophobic tail show great promise for the future Acknowledgements Dan Lundberg at Chalmers University of Tech- nology is acknowledged for help with the literature search. This review on natural surfactants and some of the work referred to are part of the activities within SNAP, the Centre for Surfactants from Natu ral Products which is sponsored by VINNOVA and by several companies. References and recommended reading 1 speci interes Se outstanding interest LU} Kosai N.Biowfactants. Surfactant Science Series 48, New York Maeol Dekker, 193, LB] Halday HL. Controversies: synthetic or natural surfactant. ‘The cae for natural surfactant. J" Perinatal Med ro924417-405, 1B] Hatebuts D, Hommel R, Claus R, Kleber H-P. Eracelar microbial pide a hisuractants A Biochem Eng/Bio Teco! 1984 33S3-93% FH] Weber L, Such J, Haufe G, Hommel R, Kleber H-P. El dation of the structure of sa unusual eject Torulopis apo. Carebydate Res 199,206 13-19. [5] Guecra-Saatos LH, Kappeli O, Fichter A. Dependence of Preuiomonas dergnoss contiauous eat iosuractant production on nutional snd eironmental factor. Appl Micobit Biotechnol 1862444348 (6) Sultan ER. Molecular genet of biosueactant production, Cure Opin sotenot 153K9:26-26, Ir} Prathi'V, Cameotra $5, Novel sucrose lipid produced by ‘+ Seria marcecon and is application in enhanced 0 0% ey. Su Det 20003833-237 Sucrose lipids wih excelent ailty to emul Hydrocarbons, lading crue ol were produced. The sfactant Was miture of ‘0 hydrowalno ae eters of sero Ts] Zuber P, Nakano MM, Marah! MA. 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Phise separation and ‘© agresute“agurezte interactions inthe COGI/CIUGL Bal Inleicoside water system. phe dagram and NMR sel- Aliusion study Langmul 198,1:6396-640, ‘The paper dnc the mechani Pehind the iguit-figuid phase separation that cert inthe rikellareepon; tis ceion pi Into one concentrated and one diute phase, both containing mi- xls. [2i}”"Nisson F, Sideman O, Johansson I. Four diferent C8G1 tylluconides, Anomeric effects and he inflcnse of righ branched iydrocarbon chaias J Callld Ineeace Set Io9s.208131-13, [22] Dowtler HD, Gopter A Lyowople laud cesta ia binary sgxtems nakylgucosdes/eater.J Dispersion Soi Technol rove.0.35-58 [23] Hanzsshe D, Schulte J. Thermotrpie and hotropie proper: lies of allsl-s-oglucopyranoside susfactats. PCCP 19999590, [24] Sierra ML, Svensson M. Mined micelles containing aliyle- cose eet ofthe chai engl and the polar head group {Langmue 199152301-2306, ‘onal surfactant was studied. A song negative value of the ite= ‘ction parameter, indatingatwactne interaction, was obtained for the mistre of slylcosie and the cationic sarfactan. A somewhat smaller negate Balu was obtained forthe miature © ayeoside surfactant with the anionic surfactant. No interaction ‘es con forthe alkypeosde-sleholethorgate mitre [BS] Holland PM Rubingh DN. Mlaod surlacant sjstems. ACS Symp Ser SO Washinton: American Chemical Society. 192. [26] Lijeyvt , Kronberg B. Comparing deyt femalonide and ‘octtetiyene gleolmono n-decyl ether in mised micelles vith dodeeylbenzne sulphonate. Formation of med mi ‘eles J Colloid Interface Set 200222139164 [27] Liljegyt , Kronberg B. Comparing decyl Bmaltoide and osutcthylene sheollmono mde! ether ia mised micelles ith dodeeylbenzene slphonate-2 Interaction of mised mi {lle wih pobwiypyrodons. J Colloid Ineelace Set don. 222165 "16 [PST Laughlin RG, Fu ¥.C, Wieko FC, Shee J, Munyon RE. “The phys science of N-dodecanoy-N-metygucumine and fs aqueous mixes in: Hoimbers K, eto. Nove! UETC- tants, New York: Mare! Deter, 199631-8. [29]. Zw ¥-P, Rosen MJ, Vinson Pk, Morel SW. Sarice proper: ‘© tes of Nalkanoy-N-meryglucamins and relted materia SF Sort Det 19992:357-362 The cflet of the numberof hyde groups in polo! surfactants ‘on pico chemical parameters sich as CMC, area per molecule and Krallt point was investigated The values of all hee parame: ters were reduced with decreasing number of hydrox groups. [30] "Alien DK, Tao BY. Carbonate alkyl eter derivatives ax ‘onus J Sut Det 199}2:38- 390 [BIT Fregapane ©, Samey DB, Vallwon EN. Face chemoenzy- nai yates of monosaccharide fatty acid ester. Bicstal- is 1811918, [52] Samey DB, Kapller H,_ Fregipane G,_Vulfon EN. CChemocnzyratie hess of disaccharide fy seid exter. J ‘Am Oil Chem Soe 1984 71-71-71. [S3] Biker UA, Matthews B,Suares H etal. Sugar fat ac exer ‘© alectnts: structure and ulimate aerobic bioderadstion. Soot Det 200051-12, A sries of sop esters with iferet sized sugar headgroups wore Jnveatigated with respect to imate aerobic rodegradation Sagar cexters with ethers sllonate oe an ety gro in a-poston ofthe faty at chn were alin inclodd. It was found that sariatons the sugar Beadgroup didnot nich alec the rate of biodeprad tion The sti sctants degraded at «lower rate [B4] Boker HA, Wiling I, Furlong DN, GriserF, Drummond C3 fo Sopa ty acid enter srfactants: binegradstion patsy 1 Suet Det 20003:13-28. “The paper dncustes biodegradation pathways for sugar eters facta. I was shown that normal nonionic sugar esters degre ‘initia eter ond fro Sugar exter surfactants wih either sulfonate ora etl Erp in erpstion ofthe fat acy hain Aegrided by another slower mechinisn, aroidation of the ay tain [55] Boker UA, Furlong DN, Greser F, Drummond CJ. Sugar fany acid ester serfctants: bisecatabed ross. J Sart Det 2005:29-32 (Bo) Boyd BY, Drommond CI, Krodkieska 1, Greser F. How ‘© chain tena, headgroup polymerization, and anomeric co figuration nem the themmotropie and iotrops iid ys talline phe behavior and the sir-watrinterfai adsoy- tion of gucosesased” surfactants. Langmuir” 200416 7399-7 The effect of ally cain length, headgroup polymerization and omer configuration ofa series of sugar esters om plscochem eal properties were investiga B11" Gao C, MilayistFureby A, Whitcombe MU, Vulson EN. fe Reginclectne sythesis of dimeric (gemini) and trimeric sigarbsed surfactant J Sart Det 19912293-32 ‘An impressive series of gemini sugar surfactant (and some wimere surfactants) were syasized by a combiotion of enzymatic and ‘organo hema routes (58) Vuln E. Enzymatic gmiess of surfactants. In: Holbers K, editor. Novel surfactants, New York: Marcel Dekker, 1998279-300. [39] Valvery R, Jauep! P. Chemo-ensymaticsytheis of amino sided sutictans, Am Ol Chem Soe II745879- 86, [40] Infante MR, Petes L, Piazo A. Nove cationic surfactants fiom arginine. in: Holmberg K, efor. Novel surfactants ‘New York Marcel Dekker, 19887-114 [SH] Pinazo A, Infante MR, lguierdo P, Solan C. Syathesis of “= gine based surfactants i highly concentrated water-int mul. J Chem Soe Perkin Trans 202 1535-1599. A highly concenzated wati-oll emulon was used a reaction ‘cd, twas demonstrated tht such emubonssbould be seen a a tema to apropi, pola solvents such a¢ dimethyformamige Sand also w microemulsions [32] ‘Capes P, Moran C, Infante MR. Enzymatic synthe of agnic-baied cationic surfactants, Biotechool Blot Iooors3335- 383 [i3] Piers E, Domingues C, Capes P, Ee P, Infante MR. Qual lative and quantitative ands of ew allyl mide arginine ssfactnts by high performance liquid chromatography and spill eletrophores. J Chromatogr A TSB9AS2-99-506, [HA] PieraF,Infante MR, Capes P. Chemo-eneymati smtesis of atgainebased gemint surfactants Biotechnol Bioeng dawon323-331K. Holmbrg / Curent Opinion Colo & Interface Since 6 2000) 148-159 159 [4S] Pinazo A, Wen XY, Peter I, Infante MR, Franses EL Aggro {ition behuvior in water of monomeric and yemin cationic Suraetnis derived from vaginine. Langmuir 20005 Freres) [46] PegiaduoS, Peres L, Infante MR. Synthesis, characterization and surface properics of I-V-L-tnptotunaeerolcthersur- facta. 3S Det 19983517- 525. ‘A now ipe af uyplophan-based sufactans wore synthesized and sharcterized with expect to CMC, surface tension and ea pt molecule atthe air-water iterfce. The tryptophane srictanis homed very low CMC vals [er] Ves MA, Macian M, Segue J, Infante MR, Vinardelt MP. Hemoltie action of anionic surfactants of the diacyl sine Iyper Canny. Deaben Pigs © Socrnot 971187115 las] Ves MA, Infante MR, Garcia E, Seve C, Maus M, Vinarell MP. Erythocyie hemolysis and shape changes ine ced by new ine dente stants, Chem Bil Inert Ios9:118-1s [49] Folncr BM, Holmberg K. Gothere Kligshog E, Berasti © K. Faty amide ethonylates: ymesis and self-assembly. J ‘Sort Det 201 in pres) ‘A sores of faty acid monoethanolamide ethonlates with varying numberof double bonds inthe ag chain and with varying number fat exyetlene group athe plat headgroup was vestigated. The jnuence ofthe amide bond and ofthe unstuations on adsompion al he airwater interface and on self-tenbly waa fnvediated Iso)” Folmce BM, Nyéa M, Holmberg K. Micelization and ad 15s sorption of a series of fay amide ethoxylaos. J Colloid Interface Soi 2 in press A series of fit aid monccthanolamie etbowlats with varying ‘umber of double bonds inthe ay chai apd wth varying number ‘of exetiyene groups inthe polar headaroup was investigated, The Jnflaence ofthe amie bond snd ofthe anstations on adsorption at hydrophohic slid srface and on micellization was inesizated by elipsometty and by NMI. ISI] Svensson M. Safactanls bated on serols and other site ‘compounds In: Holmberg K, editor. Novel slats. New ‘York: Maccl Dekker, 19817920, [52] Folmer BM, Sterol surfactants; from sythsss to apps: tions Ir: Kars DR. editor. Annual surfactant reviews, Vol. 3, Shell, UK Sheild Academie Pres, 20 in pros) [53] Folmer BM, Stes M, Holmberg K, Brown W. The physic ‘© chemi behavior of pytsterot ethos. J Colloid In terface 81 199:213.112-120 A series of ero etrslates as investigate, Iwas found that the tine to reach equim suctace tension was very log. ce than ‘wo hours This probly sells both exchange reactions atthe furface and a slow reorientation of molecule atthe st-water itertce. [5s] Folmer BM, Nydén M, Structure and dypamis inthe mis: lar and eubic pase ofan etholated phytescrl surfactant angie (submited. [SS] Folmer BM. Ph.D. Thesi Royal Insite of Techaoloy, Stockholm, Sweden, 200