Bioresource Technology 232 (2017) 398-407 Contents lists available at ScienceDirect Bioresource Technology | ELSEVIER journal homepage: www.elsevier.com/locate/biortech Review Recent advances and industrial viewpoint for biological treatment Mew of wastewaters by oleaginous microorganisms Chao Huang“, Mu-Tan Luo™“, Xue-Fang Chen, Lian Xiong *"*, Xiao-Mei Li*“, Xin-De Chen*°“* 4015 Key Laboratory of Renewable Berg, Cugow 510610, 8 China Gung institut of Emery Conversion Chinese Academy of Snes Cuneo 510610, PRChina ‘Guorgiong Provincial Key Laboratary of New ane Renewable Eery Resear an Developmen. Guangzhou 510640 PR China ‘inert of chinese Aeiemy of Sones Being 10008, PRChina HIGHLIGHTS ‘ Biological treatment of wastewaters by oleaginous microorganisms is reviewed. {the potential of industilization ofthis technology is dacusse {The issues forthe industrial application ofthis technology are presented + Critical outlook for development of this technology is given. ARTICLE INFO ABSTRACT ‘arc ion Recently, technology of using oleaginous microorganisms for biological ieatment of wastewaters has Received 29 December 2016 become one hot topic in biochemical and environmental engineering for its advantages such 35 easy ced nese form 13 Fay 2017 for operation in basic bioreactor, having potential to produce valuable bio-products.eicient wastewa> cote 15 Februry 201 {ers treatment in short petod, et. To promote its industrilization, this article provides some compre~ Lue eet hensive analysis ofthis technology such as its advances, issues, and outlook especialy from industrial Viewpoint. In dei the types of wastewaters en be reed and the Kinds of olen mirooganisns ew cater tmaimect sed for biological treatment are itradced the pote o instal appicton and sss eltvely oe low COD removal, low lipid yield, cost of operation, and lack of scale up application) of this technology. ‘tral pple are presented, and some critical outlook mainly on co-culture method, combination with other treat- Covent ‘ents, process controlling and adjusting are discussed systematically. y tis article, some important Comtination with ther eaten Information to develop this technology can be obtained, Proces contraling and adjusting © 2017 Elsevier Lid ll rights reserved, contents 1, ntoduction .. cose essessressneereessnsriensseressretseessntanensesesseiessssrsss 399 2 Wastewaters treated by oleaginous microorganisms... ee 2.1, Treatment of wastewaters from food industry 399 22, ‘Treatment of municipal wastewater or sludge ee 400 233, Treatment of fermentation wastewaters.» mo mo 2 ct 2 400 3, Oleaginous microorganisms used for biological treatment of wastewaters... - 22 aon 3.1. Oleaginous yeasts 401 32, Oleaginous microalgae «.....- : es 401 33, Oleaginous fungi and bacteria «0. 2 aoa 4. Potential for industrial application of wastewaters treatment by oleaginous microorganisms and the main issues ofthis technology... 402 4.1. Potential for industrial application of wastewaters treatment by oleaginous microorganisms 02 42, issues of wastewaters treatment by oleaginous microorganisms 402 * Corresponding author at: No. 2 Nenayan Rod, Tianhe Distt, Guano 510640, PRChina mal adr etic (X-D, Che}. hp:libdiorg)10.10165bireen.2017.02.055. (09508524) 2017 see I. Ig reserveHuan reso Techy 222 (2017) 298-407 389 42.1. Low COD removal after biological treatment. az 42.2. Low lipid yield of oleaginous microorganisms 402 423. Cost of operation : 403, 424. Lack of large scale application 403 5. Outlook. 403 5.1. Concur of microorganisms 403, S.11-Co-culture of microorganisms in ciferet fields - 403, 51.2. Co-eulture of oleaginous microorganisms for biological ieatment of wastewaters 403, 5.13. _Co-eulture of oleaginous micoorganisms and other microorganisms for biological treatment of wastewaters 404 5.2. Combination with chemical or physical treatment. 04 53. Controling and adjusting the fermentation process ‘04 6. Conclusion 405; ‘Acknowledgements - = 405 References 405 1. Introduction including the types of wastewaters can be treated and the kinds In modern society, wastewaters are always one critical problem. in city organization and industrial manufacture, Biological treat- ment of wastewaters is usually applied for many urban or indus~ trial wastewaters in the field of food, fermentation, papermaking, and so on. For biological treatment of wastewaters, aerobic diges- tion and anaerobic digestion are two main technologies that both have a long history (Chan etal, 2009). Usually, these technologies require some special equipment such a5 UASB, EASB, etc. (Coluss et al. 2009; Wei etal, 2071), and thus the upfront investment is usually high. Recently, wastewaters treatment by the oleaginous ‘microorganisms attracts much attention for its many advantages: ‘one the hand, this technology can be carried out in basic bioreactor and fermenter (the bioreactor and fermenter which can be used for ‘common fermentation) and thus requires little upfront invest- ‘ment; on the other hand, this technology can not only treat the ‘wastewaters efficiently within short period but also can generate some valuable bio-products (Fig. 1). From the past decades, various research articles reported the technology of using oleaginous microorganisms for biological treatment of wastewaters, but few articles gave some comprehensive analysis ofthis technology such asits advances, issues, and outlook especially from industrial view- point. In this review article, the advances of wastewaters treat- ‘ment by oleaginous microorganisms are introduced firstly Wastewaters Food industry wastewaters Municipal wastewater (oF sk Fermentation wastewaters iz) of oleaginous microorganisms used for biological treatment, then the potential of industrial application and the issues of this tech- nology such as relatively low COD removal, low lipid yield, cost ‘of operation, and lack of scale up application are presented. Finally, some critical outlooks of this technology including the application of co-culture, combination of chemical or physical methods, and controlling and adjusting the fermentation process are discussed systematically. Overall, this review article can offer important information to develop the technology of using oleaginous microorganisms for biological treatment of wastewaters. 2. Wastewaters treated by oleaginous microorganisms. In recent research articles, food industry wastewaters, munic- pal wastewater (or sludge), and fermentation wastewaters are the three main types of wastewaters which can be treated by ‘leaginous microorganisms (Table 1). In this part, the details for ‘these bio-pracesses are introduced systematically. 2.1, Treatment of wastewaters from food industry In modern society, food industry is important. Many food pro- cessing such as olive oil-producing industry (Paraskeva and Basic Wastewaters treatment (Effient in short period) bioreactor or fermenter Valuable products generation feria ysnccharides, citric acid, microbial biomass, et.) il, clorophyl, eaten, (Oleaginous microorg: ‘Oleaginows yeasts Oleaginous algae Oleaginous fungi Oteaginous bacteria Fe. Techanogy of sing leaginous sierorganiams for bilgi treatment of wastewaters and it advantages00 Huang to /Besure Tecnology 22 (2017) 298-407 Bloc! weatment of ome wastewaters by leginos mireorsanisns Wastewater Micoonanis COD removal) Reference ‘Wastewaes fom ood indir) Monosadium ghitamate wastewater Rhootoa his 5st ue a (2006) ‘lve ai ai wastewater Lipomyees stark Nev Yousut ea. (2010) Potato processing wastewater ‘Aspe ore maz Mania et al (2013) Seyean processing wastewater Caer pyrene ma ongyang eta (2017) Steh wastewater Rader tins © Xue ett (2010) Manica wsteater sadge Sewae sludae Lpomyees stares Na Angebauer ta. (208) Paper ml sludge (ypiecocas vise Na Deeba ct al 2016) Manipal wastewater Rader hts, baespriun raids and Cyplcocs uvetus NA Pozs et a (2013) Municipal wastewater Covealture of olaginous yeast ad algae 3s (hit al (2011) ‘Municipal wastewater (lesginous consortia (eas aod bacteria) a Halle al. (2011) Fermentation wasters ‘Brewery etMuees oder tins Na Schneider et ab. (2013) Boeenanol wastewater todesporium trues ns Zhou eta. (2013) Disiery wastenater ypeorocus curves 80 ‘Gonzale- Gari et al (2013) Disilerywastenster odor hatin Bt (Gonzale-Garia et (2013) [AE fermentation wastewater nes creme 80 (Chen eal. (2012) [ABE Fermentation wastewater ‘chosporondermats 82 Penge 2013) [ABE fermentation wastewater ‘ichesporon ewoneun 30 Xiong el. (2013) Fermentation event in Ribotlovin (82) Calls preoidsa 92 Sun ea (2013) T Wa means nor avaiable Diamadopoulos, 2006), starch industry (Feld etal, 1987), monaso- «dium glutamate industry (Liu et al, 2007), ete. will generate great rhumber of wastewaters. Usually, wastewaters from food industry has high BOD;/COD (high fermentability) and thus can be treated by traditional anaerobic digestion (Hassan and Nelson, 2012). Recently, many researches showed that oleaginous microorgan- isms can also be used for the treatment of wastewaters from food industry Firstly, oleaginous yeast Rhodotorula glutinis was used for ‘microbial oil production in monosodium glutamate wastewater that the COD removal could reach 85.51%, but the lipid yield was ‘ot high (less than 0.25 g/L) (Xue et al, 2006). However, the lipid yield could increase greatly by adding glucose to monosodium glu- tamate wastewater with the strategy of glucose feedback addition (Xue et al, 2008), For another, oleaginous yeast Lipomyces starkeyi ‘was used to treat olive oll mill wastewater and generate lipids suit- able for biodiesel production (Yousuf et al, 2010). In addition, ‘leaginous filamentous fungus Aspergillus oryzae was used to treat potato processing wastewater that the removal of COD was 74.2% ((Muniraj et al, 2013). Moreover, itis found that soybean process- jing wastewater could be used for the cultivation of oleaginous microalga Chlorella pyrenoidosa and the COD removal was close to 80% (Hongyang et al, 2011). Additionally, starch wastewater was used as raw material for microbial lipid production by Rhodo- ‘orula glutinis in pilt-scale and this initially showed the potential of industrial application of using oleaginous microorganisms for ‘wastewaters treatment (Xue et al, 2010). 22. Treatment of municipal wastewater or sludge ‘As the increase of human population and development of indus- tuy, the treatment of municipal wastewater attracts more and more attention. Traditionally, anaerobic-aerobic treatment is the basic method for the treatment of municipal wastewater (Chat et al, 2008). At the same time, sewage sludge, the residue produced dur- ing the process of wastewater treatment, its treatment becomes another challenge in wastewater management (Fytili and Zabaniotou, 2008). In the recent years, oleaginous microorganisms were proven to be possible to treat the municipal wastewater or sludge. For example, oleaginous yeast Lipomyces starkeyi was used to teat sewage sludge and accumulate microbial lipid that the lipid content could be as high as 68% (Angerbauer et al, 2008). Besides sewage sludge, itis also found that paper mill sludge can be converted into neutral lipids by oleaginous yeast Cryptococcus vishniacci for biodieset production (Deeba et al, 2016) On the ‘other hand, oleaginous yeasts such as Rhodotorula_glutinis, ‘Rhorosporidium foruoides, and Cryprococcus curvatus, could be used for municipal wastewater treatment (Pirozzi et al 2013) For the treatment of municipal wastewater, co-culture of various microor- -ganisms such as oleaginous yeast and algae could be one efficient strategy (Chi et al, 2011). Also, nine oleaginous yeasts and one ‘leaginous bacterium were used for municipal wastewater treat- ‘ment and the COD removal was approximately 81% (Hall et al, 2011). 23. Treatment of fermentation wastewaters Fermentation is one important biological technology that can synthesize various valuable compounds to satisly the requirement ‘of chemical industry, energy. food, etc. However, various wastew= aters are usually generated after fermentation. Generally, the bio- logical teatment of fermentation wastewaters usually uses _anaerobic-aerobic method by activated sludge with some special ‘equipment (Lu et al, 1999: Pant and Adholeya, 2007), In the past ‘years, itis also found that oleaginous microorganisms can treat the wastewaters from diferent fermentation industries, Ethanol fermentation will generate large amount of wastewater especially after recovery of the products from fermentation broth by distilla- tion. This wastewater was shown as one promising substrate for ‘leaginous microorganisms. For instance, brewery effluent was used as fermentation substrate to accumulate microbial oil and ‘carotenoid simultaneously by oleaginous yeast Rhodotorula glutinis (Schneider etal, 2012), For another, oleaginous yeast Rhodasporid- ium toruloides was used to treat bioethanol wastewater, and the COD removal, yeast biomass. lipid content could reach 72.3%, 38 g/L, and 349%, respectively (Zhou et al, 2013) In addition, ‘oleaginous yeasts Rhodotorula glutinis and Cryptococcus curvatus ‘were used to treat distillery wastewater, and the COD removal ‘could reach 80% (Gonzalez-Garcia et al, 2013), Similar to ethanol fermentation, ABE (acetone-butanol-ethanol) fermentation also ‘generates great amount of wastewater after eecovery of theHuan reso Techy 222 (2017) 298-407 401 leasinos micrerganiss sed for wastewaters Weatenent Mirooranimn Sabre Biomass fa) Upid coment Reference D o ‘tages yeasts Riodotorla tins Browory effluents sx as Schneier et a. (2013) Rbodoorla ‘Olive mil wastewater 961 Na Jarbou eta 2013), ‘mucin Tchesporen tans ABE femeaation wastewter 49 uaz ‘ong (2015) Tcheporen derma ABE fermentation wastewater fa 5 Penge a. (2013) Tickespron ABE fermentation wastewater 58 94 ‘Chen ea. (2012) ‘oremiferme Ccyprctcs cura Disilery wastewater 519 252 Gonaaler- Gari a (013) Coyprecocusabigus Pulp and paper ml unt ws Na Singhal and Thakur 2012) Copter tment Winery wastewater na NA Santos eta 2014) Copter pedslcs Pulp sl wastewater na NA Fernandes et (2014) iodespordum Distlery wastewater a B55 Lng a (2013) tories Lipomyeesstarkey 239 a8 Lie a. (2013) Yaron lipayice Olive wil wastewater based media 70 va Suse a. (2011) teazneus mirootee Cae vugars Atco wastewater 009 2 Feng et al. (2011) Chive protaiecsides Thocanate wastewater 13 S06 Ryvet al (2014) Cire pyrenadosa gery wastewater hose t003 oseto 18 Wang eal 2012) Cire eipsoien Domestic secondary event ows 8 Yang eta (2012), Secmedesmusacuis Manipal wastewater his 283 ‘de Ava et (2013) ‘Scenedesmus guns Fishpond (FP) ducge, and mull secondary seting tank (MST) 2.0 525 Mada sa Malick (2011) ddschages SconedesrusBiuzo Food wastewater eluent 149 3505 Shine ab. (2015) Neacors Anaerbialy digested dairy manare Ghoveto 10 188 levine etal (2012) uryocreis bron Nitatevch wastewter supplemented with CO; 226 203 ‘Yeesang and Cheieslp (01a) Arties persis Olive mill wastewater 70 1691 Markou ea. (2012) Olggnes fies and bacteria ‘Asperais onze Pkatoprocesing wastewater Atco cnet Trchoderma reeset Riodecrs pacis odococes 5p. HAL ‘wastewaters fom equatization tank ‘wastewaters fom equalization ark Primary effluent wastewater Chenithemomechancal pulping eluent NA (Highest iid yield was Muni) eta. (2013) 35x os 2a Bhan ee a. 014) O58 fanz Bhan et aL 2014) NA NA Malle a. (2011) nA NA Duet (2tt) © NA means not avaible products from fermentation broth by distillation. It is also shown ‘that oleaginous microorganisms can treat the ABE fermentation ‘wastewater that the COD removal was close to 70% (Chen et al 2012; Peng et al, 2013; Xiong et al., 2015), Other wastewaters from fermentation can also be treated by oleaginous microorgan- sms. For instance, oleaginous microalga Chlorella. pyrenoidosa ‘was used to treat fermentation effluent in Riboflavin (B2) and the COD removal was close to 90% (Sun et al, 2013). 3. Oleaginous microorganisms used for biological treatment of wastewaters Generally, various microorganisms including, yeasts, algae, fungi, and bacteria could accumulate lipid to 20% of their dry weight, and so called oleaginous microorganisms (Ratledze, 1991). To date, all kinds of these microorganisms have been uti- lized for biological treatment of different wastewaters (Table 2) and the details are introduced in the following parts 3.1 Oleaginous yeasts Recently, many oleaginous yeasts have been applied for micro- bial oll production due to their excellent capacity for substrate con- sumption, lipid production, fast growth rate, easy for culture, stable yield etc (Sitepu et al, 2014), For the biological treatment fof wastewaters, the oleaginous yeasts used mainly includes the yeasts from genus of Rhodororuia such as Rhodotorua glutinis (Cheitsitp et al, 2011; Chi et al, 2011; Schneider et al, 2013; Xue et al, 2008) and Rhodotorula mucilaginasa (Jarboui et al, 2012, 2013), genus of Trichosporon such as Trichosporon cutaneum (Dragigevic et al, 2010; Xiong et al, 2075), Trichosporon dermatis (Peng et al, 2013), and Trichosporon coremiiforme (Chen et al, 2012}, genus of Cryptococcus such as Cryptococcus curvatus (Chi ft al, 2011; Gonzalez-Garcia et al, 2013), Cryptococcus albidus (Singhal and Thakur, 2012), Cryprococcus laurentii (Santos et al, 2014), and Cryptococcus podzolicus (Feinandes et al, 2014), and some other kinds of oleaginous yeasts such as Rhodasporidium toruloides (Ling et al, 2013; Zhou et al, 2013), Lpomyees starkeyi (Angerbauer et al, 2008; Huang et al, 2011; Liu et al, 2013), Yar- 1owia lipolytica (Lan et a, 2008; Lanciott etal, 2005; Sarts et al 2011) and so on. 3.2, Oleaginous microalgae For a long time, oleaginous microalga is considered as one ‘important oleaginous microorganism for bio-fuel production (Lam and Lee, 2012). In spite that oleaginous microalga seldom uses some special substrates such as lignocellulosic hydrolysates {or fermentation (Huang et al, 2013), but it has many advantages ‘which make it has great potential for wastewaters treatment. Com- ppared with yeast, mictoalga can survive in the wastewaters with low nutrient concentration due to its autotrophic character (Cai402 Huang to /Besure Tecnology 22 (2017) 298-407 et al, 2013). More specially it requires litle aseptic environment ‘and thus can be operated in the “open-door” environment (Ratledge and Cohen, 2008). In addition, many by-produets such 4s f-carotene and algal polysaccharide with high value can also be synthesized by different microalgae (Raja et al, 2007; Zhu ‘et al, 2014), Although the growth period of microalgais relatively longer when compared with other microorganisms and its collec- tion is also one industrial issue (Barros etal, 2015), it still has great potential for industrialization for considering its above excellent ‘capacity. For the biological treatment of wastewaters, the oleagi- nous microalgae used mainly includes the microalgae from genus ‘of Chlorella such as Chlorela vulgaris (Feng et al., 2011; Ji et al, 2013), Chlorella protothecoldes (Ryu et al, 2014), Chlorella pyrenot- ddosa (Wang et al. 2012), and Chlorella elipsoidea (Yang, et al, 2011), genus of Scenedesmus such as Scenedesmus acutus (de Alva et al 2013), Scenedesmus obliquus (Mandal and Mallick, 2011), ‘and Scenedesmus bijuga (Shin ef al, 2015), and some other kinds $f oleaginous microalgae such as Neochloris oleoabundans (Levine et al, 2011), Botryococcus braunié (Yeesang and Cheirsilp. 2014), Arthrospia platensis(Natkou et al., 2012) and so on. 33, Oleaginous fungi and bacteria ‘Compared with microalgae and yeasts fungi and bacteria were applied relatively less in biological treatment of wastewaters However, these microorganisms could generate many special products which yeasts and microalgae seldom accumulate (Sancholle et al, 2004; Waltermann et al, 2005) and thus they hhave special application in industry. To date, the oleaginous fungi and bactetia applied for biological treatment of wastewaters ‘mainly include Aspergillus oryzae (Muniraj et al, 2013), Mucor Circinelloides and Trichoderma reeset (Bhanja et al, 2014), Rhodococ- ‘us opacus (Hall etal, 2011, Rhodococcus sp. (Du et al 2011) and 4. Potential for industrial application of wastewaters treatment by oleaginous microorganisms and the main issues of this ‘technology 4.1. Potential for industrial application of wastewaters treatment by ‘leaginous microorganisms Basically, the industrialization of biochemical technology requires many factors (Saxena et al, 2009), The substrate is the ‘most critical one that a suitable substrate for industrial biochemi- ‘al technology should be low-cost and available (Huang et al, 2013), Wastewater is usually wastes from industry, agriculture or city, and thus its cost is extremely low. In addition, various wastewaters exist from different place, thus it has great availabil- ity. Therefore, wastewater can be considered as one promising sub- strate for industrial biochemical technology. Besides substrate, the ost of bioconversion i also important. The biological treatment of ‘wastewaters by oleaginous microorganisms merely requires a short fermentation. period (usually several days), and moderate Temperature (usually room temperature is suitable}, and thus the ‘energy consumption ofthis technology is small. The basic fermen- tation equipment could be used for this bioconversion and there- fore it requires no extra and expensive building of new ‘equipment. In addition, the floor area ofthis technology is less than that of traditional biological treatment. Overall, the cost of this bio- ‘conversion is suitable for industrial application. Usually, the downstream technology plays an important rote in biochemical engineering. For the biological treatment of wastewa- ters by oleaginous microorganisms, the downstream technology mainly includes two parts, namely the biomass harvesting and collection, and microbial lipid recovery. For biomass harvesting ‘and collection, both centrifugation and flocculation can be applied (Christenson and Sims, 2011), For lipid recovery, many methods for cell disruption (ultra-sonication, high-pressure homogeniza- tion, French pressing. bead milling, microwave, chemical lysis, ‘osmotic shock) and lipid extraction (organic solvent extraction, supercritical fluid extraction, ultrasound-assisted extraction microwave-assisted extraction) are used widely (Halim et al, 2012), Itis worth noting that all these technologies should avoid high temperature and pressure to reduce cost in industrial application. Considering the cost from upstream to downstream technology, the biological treatment of wastewaters by oleaginous microor- 4ganisms has great potential for industralization. To date, some pilot scale researches have been carried out forthe biological treat- ment of wastewaters by oleaginous microorganisms (Fernandes cet al, 2014; Xue et al, 2010; Zhu et al, 2013) and the information is critical forthe industrial application because it proves the possi- ilty of using this technology in large scale. n future, more appli- ‘ation should be carried out in various industries 4.2 Issues of wastewaters treatment by oleaginous microorganisms 42.1. Low COD removal after biological treatment ‘AS one novel technology for biological treatment of wastewa- ters, although it has a relatively short period for treatment, the COD degradation of biological treatment by oleaginous microor- -ganisms is still usually lower than that of traditional technologies ‘of both aerobic and anaerobic ones. In fact, by the traditional anaerobic-aerobic biological treatment, the total COD removal of industrial and municipal wastewaters can be higher than 80% and even higher than 90% (Chan et al, 2009), And in many situa- tions, the biological treatment of wastewaters by oleaginous microorganisms had a relatively low COD removal and even lower than 80% (Table 1). Undoubtedly, relatively low COD removal of this technology makes it Iess competitive for industrial application. ‘Therefore, to full the industrialization of wastewaters treatment by oleaginous microorganisms, the most important mission is to increase the COD removal ofthis technology to satisfy the require- ‘ment of water standard. 422. Low lipid yield of oleaginows microorganisms Compared with traditional lipid fermentation on different ‘media containing high concentration of fermentation substrates (Huang etal, 2013; Sitepu et al, 2014), the biomass and lipid con- {ent of oleaginous microorganisms in different wastewaters are simall (even less than 2.0 g/l and 20%, respectively, see Table 2) and thus the lipid yield of oleaginous microorganisms is low. If the microbial oil is the final products, the economic value of this bioconversion is undoubtedly less competitive. The unsuitable C) N ratio of wastewaters for lipid synthesis might explain this phe- ‘nomenon (Papanikolaou and Aggelis, 2011). To increase the lipid yield of oleaginous microorganisms in wastewaters, more ‘exogenous carbon sources could be added into different media (Huang et al, 2011; Xue et al, 2008), but this will also increase the cost of substrate. Thus, unless the method to increase the lipid yleld of oleaginous microorganisms could be found from either ‘genetic or engineering viewpoint, the better idea is stil explore the value of by-products of lipid fermentation. In fact, besides ‘microbial lipid, many other valuable products such as chlorophyll (Farooq et al, 2013; Li et al, 2008), carotenoid (Saenge et al, 2011; Schneider et ai, 2013), polysaccharides (Xiong et al, 2016; Zhu et al, 2014), citric acid (Lanciotti et al, 2005; Sarrs et al, 2011), and even the biomass of oleaginous microorganisms (Zhveng et al, 2005) might also be generated during the biological treatment of wastewaters by oleaginous microorganisms. Thus, it