SUPPLEMENTARY MATERIALS

Title:

Enhanced oil production by the tropical marine diatom Thalassiosira sp. cultivated in outdoor photobioreactors

The names of authors:

Pintaka Kusumaningtyasa, Santi Nurbaitia,*, Gede Suantikab, Muhammad Bachri Amranc,

Zeily Nurachmana,**

The affiliations and addresses of the authors: aBiochemistry Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi

Bandung, Jl. Ganesha 10, Bandung 40132, Indonesia bMicrobial Biotechnology Research Group, School of Life Sciences and Technology, Institut

Teknologi Bandung, Jl. Ganesha 10, Bandung 40132, Indonesia cAnalytical Chemistry Division, Faculty of Mathematics and Natural Sciences, Institut

Teknologi Bandung, Jl. Ganesha 10, Bandung 40132, Indonesia

*Dedicated to the memory of Dr. Santi Nurbaiti (†29.05.2016) Fig. S1. Simple air lift-photobioreactors used to evaluate the growth of Thalassiosira sp. in agricultural fertilizer-based media under indoor conditions. The photobioreactors were made from transparent bottles with a height of 25 cm, an external diameter of 9 cm, and a working volume of 800 mL Fig. S2. Vertical column photobioreactors used to evaluate the growth of Thalassiosira sp. outdoors. Photobioreactors were made from transparent acrylic material with a diameter of 15 in, a height of 1 m, and a working volume of 17 L Fig. S3. Algal biomass: (A) wet biomass (pasta), and (B) dry biomass Fig. S4. Crude lipid extract from algal biomass Fig. S5 Accumulation of intracellular neutral lipids in Thalassiosira sp. cells cultivated outdoors. The oil accumulation was monitored using Nile red fluorescence Table S1. Biomass and oil productivity of Thalassiosira sp. cultured in four different

media

Medium Biomass productivity (mg Oil content Oil productivity (mg L–1 d–1) (% DW) L–1 d–1) AF3 27.48 ± 1.23 37.68 ± 1.83 10.37± 0.89 Nitrate-ES 27.84 ± 2.54 24.19 ± 3.26 6.71 ± 0.89 Urea-ES 20.59 ± 1.17 28.25 ± 2.34 5.82 ± 0.65 Urea-TM-ES 20.09 ± 1.06 37.98 ± 1.76 7.64 ± 0.72 Data are mean ± standard deviation of triplicates Tabel S2. Fatty acid composition of Thalassiosira sp. cultured outdoor in different

weather conditions

Season Types of fatty acids Rainy Dry Saturated (SFA): C12:0 (Lauric acid) – 1.39 ± 0.78 C13:0 (Tridecanoic acid) – – C14:0 (Myristic acid) 9.35 ± 0.87 11.39 ± 4.17 C15:0 (Pentadecanoic acid) 0.36 ± 0.02 0.68 ± 0.33 C16:0 (Palmitic acid acid) 12.13 ± 4.75 14.82 ± 2.67 C17:0 (Heptadecanoic acid) 3.15 ± 0.10 5.45 ± 1.79 C18:0 (Stearic acid) 1.47 ± 0.62 4.52 ± 0.79 C20:0 (Arachidic acid) – – C21:0 (Heneicosanoic acid) 0.37± 0.01 – C22:0 (Behenic acid) 1.06 ± 0.17 1.00 ± 0.01 C23:0 (Tricosanoic acid) 0.25 ± 0.02 – C24:0 (Lignoseric acid) 0.40 ± 0.07 0.75 ± 0.26 Total SFA 28.55 40.00 Monounsaturated (MUFA): C14:1 (Myristoleic acid) 0.38 ± 0.04 0.37 ± 0.53 C15:1 (Pentadecenoic acid) 0.31 ± 0.26 0.60 ± 0.15 C16:1 (Palmitoleic acid) 31.72 ± 3.24 35.00 ± 5.07 C17:1 (Heptadecenoic acid) 0.25 ± 0.23 – C18:1n9c (Oleic acid) 2.07 ± 1.63 1.17 ± 0.24 C18:1n9t (Elaidic acid) 3.08 ± 0.38 4.29 ± 1.12 C20:1 (cis-11-Eicosenoic acid) – 0.14 ± 0.19 C22:1 cis (Erucic acid) 1.93 ± 0.28 – C22:1 trans (brussidic acid) 0.64 ± 0.05 – Total MUFA 40.39 41.57 Polyunsaturated (PUFA): C16:3n3 (Hexadecatrienoic acid) 8.90 ± 2.66 4.54 ± 3.66 C18:2n6c(Linoleic acid) 5.13 ± 0.22 2.15 ± 0.56 C18:2n6t (Linolelaidic acid) 0.67 ± 0.12 0.43 ± 0.06 C18:3n3 (-Linolenic acid) 0.46 ± 0.09 – C18:3n6 (-Linolenic acid) 0.78 ± 0.36 – C20:4n6 (AA) 0.76 ± 0.94 0.60 ± 0.14 C20:5n3 (EPA) 10.94 ± 3.73 0.97 ± 0.68 C20:3 (Eicosatrienoic acid) 0.83 ± 0.68 9.16 ± 1.78 C20:2 (Eicosadienoic acid) 0.19 ± 0.16 – C22:2 cis (Docosadienoic acid) 1.05 ± 0.17 – C22:2 trans (Docosadienoic acid) 0.64 ± 0.30 – C22:6n3 (DHA) 0.92 ± 0.13 0.58 ± 0.00 Total PUFA 31.26 18.42