Microbial consortia: a critical look at microalgae co-cultures for enhanced biomanufacturing

Figures & data

Figure 1. Communication within microbial communities. Metabolite exchanges (arrows) facilitate various modes in which microorganisms (geometrical shapes) exhibit intra- or inter-species interactions. Communication is used for (A) quorum sensing and defining the abundance of each species and (B) type of symbiosis and roles played by partners, such as in (C) protection and (D) nutrient acquisition and division of labor. Further to this, as the community evolves, so does the communication, with the effect of causing changes to the microbial communities that are part of it, for example, by recruiting new partners (E) or by evolving existing members (F).

Table 1. Microbial co-cultures in bio-production.

			Reported product yield/concentration
Product	Reported organisms	Mode	Monoculture		Co-culture	References
Acetate	Weissella confusa 11GU1 P. freudenreichii JS15	Fermentation at 1:1 culture ratios	0.08g/kg	0.09g/kg	0.5g/kg	[83]
Astaxanthin	Haematococcus pluvialis, Phaffia rhodozyma AS2-1557	Gas Exchange: CO2 and O2 3g/L of glucose	3.68mg/L	1.09mg/L	12.95mg/L	[5]
Biomass	Haematococcus pluvialis, Phaffia rhodozyma AS2-1557	Gas Exchange: CO2 and O2 25g/L of glucose	0.62g/L	5.02g/L	5.70g/L	[5]
	Scenedesmus obliquus, Institute of Hydrobiology Candida tropicalis, Institute of Microbiology	Direct mixing, 3:1 ratio	3.5g/L	n.d.	4.38g/L	[145]
	Isochrysis galbana 8701 Ambrosiozyma cicatricosa	Direct mixing, 1:1 ratio	1.17g/L	0.31g/L	1.32g/L	[143]
	Spirulina platensis UTEX 1926 Rhodotorula glutinis 2.541	Direct mixing, 2:1 ratio	0.20g/L	1.7g/L	3.6g/L	[156]
	Chlorella vulgaris TISTR 8261 Trichosporonoides spathulata	Direct mixing	0.75g/L	10.23g/L	12.2g/L	[77]
	Chlorella sp. KKUS2 Toluraspore YU5/2	Direct mixing	1.933g/L	8.333g/L	8.010g/L	[157]
	Chlorella sp. KKUS2 Toluraspore Y30	Direct mixing	1.933g/L	8.267g/L	8.733g/L	[157]
Carotenoids (β-carotene, torulene, torularhodin)	Rhodutola glutinis DBVPG 3853, Debaryomyces castellii DBVPG 3503	Fed-batch system with co-culture 1:1 ratio	5.3mg/L, batch co-culture	8.2mg/L		[85]
EPS	Weissella confusa 11GU1 P. freudenreichii JS15	Fermentation at 1:1, with 15% w/w added flour	n.d.	1g/kg	1.52g/kg	[83]
	Agaricus blazei LPB03, Chlorella vulgaris LEB106	Direct mixing, 1:1 ratio	4g/L	0.95g/L	5.17g/L	[69]
2-keto-L-gulonic acid	Gluconobacter oxydans, Ketogulonicigenium vulgare	Fermentation with gene manipulation	n.d.	n.d.	76.6g/L (89.7%)	[158]
Propionate	Weissella confusa 11GU1 P. freudenreichii JS15	Fermentation at 1:1 culture ratios	1.15g/kg	0g/kg	0.59g/kg	[83]
Lipids	Chlorella pyrenoidosa FACHB-9 Rhodospiridium toruloides AS2.1389	Wastewater, co-culture 1:1 ratio	3g/L	3.4g/L	4-4.6g/L	[159]
	Spirulina platensis UTEX 1926 Rhodotorula glutinis 2.541	Direct mixing, 2:1 ratio	0.013g/L	0.135g/L	0.467g/L	[156]
	Chlorella vulgaris TISTR 8261 Trichosporonoides spathulata JU4-57	Direct mixing	4.14g/L	n.d.	5.74g/L	[77]
	Chlorella sp. KKUS2 Toluraspore YU5/2	Direct mixing	0.052g/L	1.141g/L	2.424g/L	[157]
	Chlorella sp. KKUS2 Toluraspore Y30	Direct mixing	0.052g/L	0.920g/L	1.564g/L	[157]

Co-cultures employed for specific products are listed, along with the organisms employed, cultivation mode and reported product yields/productivity/concentrations in mono and co-cultures. The monoculture data provided lists the yield/concentration of the primary partner (A) followed by the secondary partner, if both organisms produce the desired product (n.d.: not determined).

Figure 2. Steps involved in constructing an artificial co-culture. A bottom-up approach is shown. The desired product is defined first (I), the microbial producers are short-listed next. This can be based on metabolite profiling or on natural associations (II). From selected candidates (III) co-cultures need to be investigated to elucidate the type of partnership (IV). The highest yielding co-culture is to be selected (V), optimized (VI) and upscaled (VII).

Table 2. A selection of high-value products derived from microalgae species as monocultures.

Bioproduct	Reported species	Reported product yield/concentration	Reference
Astaxanthin	Chlorella zofingiensis ATCC 30412	10.3 mg/L	[160]
	C. zofingiensis, CCAP 211/14	0.1 pg/cell	[161]
	Haematococcus pluvialis LB 16	91.7 pg/cell	[162]
	H. pluvialis 26	40.25–51.06 mg/L	[163]
	H. pluvialis, 34/7	2.7% dry wt	[164]
β-carotene	Dunaliella salina, Sambhar Salt Lake	4.21 pg/cell	[122]
	D. salina - 19.3	7.05–8.26 pg/cell	[165]
	D. salina SAG 42.88	3.99 pg/cell	[123]
	D. salina, CONC-007	72.7 pg/cell	[166]
	D. salina, CCAP 19/18	31.6 pg/cell	[166]
	D. salina, Urmia Lake isolate	8.94–11.4 pg/cell	[167]
	D. salina, KU01	56.25 pg/cell	[168]
	Dunaliella bardawii – KU01	52.91 pg/cell	[168]
	D. salina, CCAP 19/18	70 pg/cell	[169]
Glycerol	Dunaliella sp, Sambhar Salt Lake	94.26 pg/cell	[122]
Lipids	Botryococcus braunii, UTEX 572	5.51–21 mg/L/d	[170]
	Chlorella vulgaris, KCTC AC10032	6.91 mg/L/d	[170]
	Scenedesmus sp., KCTC AG20831	20.65–39 mg/L/d	[170]
Lutein	Chlamydomonas acidophila	20 mg/L	[171]
	Muriellopsis sp., isolate from Empordámarsh	1.4–0.8 mg/L/d	[135]
	C. zofingiensis CCAP 211/14	4 mg/g dry wt	[161]
Phycobilin	Nostoc muscorum	0.0229% p/v	[134]
	Gloeotrichia natans	0.21 g/L	[129]
Phycocyanin	Galdieria sulphuraria 074G	8–28 mg/g dry wt	[172]
	Spurilina platensis	46% w/w	[71]
	S. pluriformis	9.6% w/w	[71]
	Nostoc sp.	20% dry wt	[129]

The species involved and reported product yields/productivity/concentration are provided in different units as reported in the references.

Figure 3. Representation of a microalgae-based consortium for biotechnological applications. A photo-illuminated bioreactor for culturing an artificially created synergistic consortium between algae, yeast and bacteria within a small-scale reactor is represented. The microalgae take up carbon dioxide and produce oxygen (through photosynthesis) that is, consumed by the aerobic bacteria and yeast, which in turn supply carbon dioxide (through respiration) to be consumed by the algae. Cell secretions and degradation will release biomolecules (vitamins, proteins, carbohydrates, nucleic acids and secondary metabolites) into the growth media. The bacteria will break these materials into simpler compounds to be consumed by all members of the consortium.

Tinzl-Malang SK, Rast P, Grattepanche F, et al. Exopolysaccharides from co-cultures of Weissella confusa 11GU-1 and Propionibacterium freudenreichii JS15 act synergistically on wheat dough and bread texture. Int J Food Microbiol. 2015;214:91–101.

 PubMed Web of Science ®Google Scholar

Dong Q, Zhao X. In situ carbon dioxide fixation in the process of natural astaxanthin production by a mixed culture of Haematococcus pluvialis and Phaffia rhodozyma. Catal Today. 2004;98:537–544.

 Web of Science ®Google Scholar

Wang R, Tian Y, Xue S, et al. Enhanced microalgal biomass and lipid production via co-culture of Scenedesmus obliquus and Candida tropicalis in an autotrophic system. J Chem Technol Biotechnol. 2015;9:1387–1396.

 Google Scholar

Cai S, Hu C, Du S. Comparisons of growth and biochemical composition between mixed culture of alga and yeast and monocultures. J Biosci Bioeng. 2007;104:391–397.

 PubMed Web of Science ®Google Scholar

Xue F, Miao J, Zhang X, et al. A new strategy for lipid production by mix cultivation of Spirulina platensis and Rhodotorula glutinis. Appl Biochem Biotechnol. 2010;160:498–503.

 PubMed Web of Science ®Google Scholar

Kitcha S, Cheirsilp B. Enhanced lipid production by co-cultivation and co-encapsulation of oleaginous yeast Trichosporonoides spathulata with microalgae in alginate gel beads. Appl Biochem Biotechnol. 2014;173:522–534.

 PubMed Web of Science ®Google Scholar

Papone T, Kookkhunthod S, Leesing R. Microbial oil production by monoculture and mixed cultures of microalgae and oleaginous yeasts using sugarcane juice as substrate. World Acad Sci Eng Technol. 2012;64:1127–1131.

 Google Scholar

Buzzini P. Batch and fed-batch carotenoid production by Rhodotorula glutinis-Debaryomyces castellii co-cultures in corn syrup. J Appl Microbiol. 2001;90:843–847.

 PubMed Web of Science ®Google Scholar

Angelis S, Noivak AC, Sydney EB, et al. Co-culture of microalgae, cyanobacteria, and macromycetes for exopolysaccharides production: process preliminary optimization and partial characterization. Appl Biochem Biotechnol. 2012;167:1092–1106.

 PubMed Web of Science ®Google Scholar

Wang E-X, Ding M-Z, Ma Q, et al. Reorganization of a synthetic microbial consortium for one-step vitamin C fermentation. Microb Cell Fact. 2016;15:21.

 PubMed Web of Science ®Google Scholar

Ling J, Nip S, Cheok WL, et al. Lipid production by a mixed culture of oleaginous yeast and microalga from distillery and domestic mixed wastewater. Bioresour Technol. 2014;173:132–139.

 PubMed Web of Science ®Google Scholar

Ip P, Wong K, Chen F. Enhanced production of astaxanthin by the green microalga Chlorella zofingiensis in mixotrophic culture. Process Biochem. 2004;39:1761–1766.

 Web of Science ®Google Scholar

Del Campo JA, Rodríguez H, Moreno J, et al. Accumulation of astaxanthin and lutein in Chlorella zofingiensis (Chlorophyta). Appl Microbiol Biotechnol. 2004;64:848–854.

 PubMed Web of Science ®Google Scholar

Choi YE, Yun YS, Park JM. Evaluation of factors promoting astaxanthin production by a unicellular green alga, Haematococcus pluvialis, with fractional factorial design. Biotechnol Prog. 2002;18:1170–1175.

 PubMed Web of Science ®Google Scholar

Zhang BY, Geng YH, Li ZK, et al. Production of astaxanthin from Haematococcus in open pond by two-stage growth one-step process. Aquaculture. 2009;295:275–281.

 Web of Science ®Google Scholar

Harker M, Tsavalos AJ, Young AJ. Autotrophic growth and carotenoid production of Haematococcus pluvialis in a 30 liter air-lift photobioreactor. J Ferment Bioeng. 1996;82:113–118.

 Google Scholar

Phadwal K, Singh PK. Isolation and characterization of an indigenous isolate of Dunaliella sp. for beta-carotene and glycerol production from a hypersaline lake in India. J Basic Microbiol. 2003;43:423–429.

 PubMed Web of Science ®Google Scholar

Pisal D, Lele S. Carotenoid production from microalga, Dunaliella salina. Indian J Biotechnol. 2005;4:476–483.

 Google Scholar

Phadwal K, Singh P. Effect of nutrient depletion on beta-carotene and glycerol accumulation in two strains of Dunaliella sp. Bioresour Technol. 2003;90:55–58.

 PubMed Web of Science ®Google Scholar

Gomez PI, Gonzalez MA. The effect of temperature and irradiance on the growth and carotenogenic capacity of seven strains of Dunaliella salina (Chlorophyta) cultivated under laboratory conditions. Biol Res. 2005;38:151–162.

 PubMed Web of Science ®Google Scholar

Rad FA, Aksoz N, Hejazi MA. Effect of salinity on cell growth and β -carotene production in Dunaliella sp. isolates from Urmia Lake in northwest of Iran. Afr J Biotechnol. 2011;10:2282–2289.

 Google Scholar

Sathasivam R, Kermanee P, Roytrakul S, et al. Isolation and molecular id entification of β -carotene producing strains of Dunaliella salina and Dunaliella bardawil from salt soil samples by using species-specific primers and internal transcribed spacer (ITS) primers. Afr J Biotechnol. 2012;11:16677–16687.

 Google Scholar

Mojaat M, Pruvost J, Foucault A, et al. Effect of organic carbon sources and Fe2+ ions on growth and β-carotene accumulation by Dunaliella salina. Biochem Eng J. 2008;39:177–184.

 Web of Science ®Google Scholar

Yoo C, Jun S-Y, Lee J-Y, et al. Selection of microalgae for lipid production under high levels carbon dioxide. Bioresour Technol. 2010;101 Suppl:S71–S74.

 Web of Science ®Google Scholar

Garbayo I, Cuaresma M, Vilchez C, et al. Effect of abiotic stress on the production of lutein and beta-carotene by Chlamydomonas acidophila. Process Biochem. 2008;43:1158–1161.

 Web of Science ®Google Scholar

Blanco AM, Moreno J, Del Campo JA, et al. Outdoor cultivation of lutein-rich cells of Muriellopsis sp. in open ponds. Appl Microbiol Biotechnol. 2007;73:1259–1266.

 PubMed Web of Science ®Google Scholar

de Morais MG, da Silva Vaz B, Etiele Greque de M, et al. Review article biologically active metabolites synthesized by microalgae. Biomed Res Int. 2015;2015:1–15.

 Web of Science ®Google Scholar

Sekar S, Chandramohan M. Phycobiliproteins as a commodity: trends in applied research, patents and commercialization. J Appl Phycol. 2007;20:113–136.

 Web of Science ®Google Scholar

Sloth JK, Wiebe GW, Eriksen NT. Accumulation of phycocyanin in heterotrophic and mixotrophic cultures of the acidophilic red alga Galdieria sulphuraria. Enzyme Microb Technol. 2006;38:168–175.

 Web of Science ®Google Scholar

Markou G, Nerantzis E. Microalgae for high-value compounds and biofuels production: a review with focus on cultivation under stress conditions. Biotechnol Adv. 2013;31:1532–1542.

 PubMed Web of Science ®Google Scholar