Isolation, screening and comprehensive characterization of candidate

microalgae for biofuel feedstock production and dairy effluent treatment: a

sustainable approach

Ashutosh Pandey1, Sameer Srivastava1, Sanjay Kumar2*

1Department of Biotechnology, Motilal Nehru National Institute of Technology Allahabad, Allahabad (U.P.), India

2School of Biochemical Engineering, Indian Institute of Technology (BHU), Varanasi (U.P.), India

*Corresponding author:

Dr Sanjay Kumar

sanjaykr.bce@iitbhu.ac.in

Table S1

Wastewater sample collection sites for microalgae isolation.

Collection site Coordinates Date of Collection

Microalgal isolates

Pond_Near Mahabirpur Teliyarganj, Prayagraj (UP), India

25°30'12.0"N 81°52'02.7"E

December, 2015

ASK01-17

Aeration tank of dairy effluent plant Shyam Dairy products, Prayagraj (UP), India

25°38'26.0" N, 81°83'80.0" E

February 2016

ASK18-28

Table S2

Correlation equations of microalgae biomass (gL-1) vs absorbance (680 nm).

Microalga isolate Correlation equations

ASK01 Biomass (gL-1) = 0.2728 × Abs680 ̶ 0.0829 (R2 = 0.9717)

ASK03 Biomass (gL-1) = 0.2368 × Abs680 + 0.048 (R2 = 0.9790)

ASK07 Biomass (gL-1) = 0.2994 × Abs680 ̶ 0.0415 (R2 = 0.9838)

ASK08 Biomass (gL-1) = 0.3304 × Abs680 ̶ 0.0493 (R2 = 0.9987)

ASK09 Biomass (gL-1) = 0.2561 × Abs680 ̶ 0.0127 (R2 = 0.9797)

ASK10 Biomass (gL-1) = 0.3607 × Abs680 ̶ 0.0196 (R2 = 0.9711)

ASK14 Biomass (gL-1) = 0.4364 × Abs680 + 0.0028 (R2 = 0.9791)

ASK16 Biomass (gL-1) = 0.7184 × Abs680 ̶ 0.0497 (R2 = 0.9984)

ASK17 Biomass (gL-1) = 0.3634 × Abs680 ̶ 0.0145 (R2 = 0.9959)

ASK18 Biomass (gL-1) = 0.2533 × Abs680 +0.0543 (R2 = 0.9878)

ASK20 Biomass (gL-1) = 0.4173 × Abs680 ̶ 0.0365 (R2 = 0.9950)

ASK21 Biomass (gL-1) = 0.369 × Abs680 ̶ 0.0228 (R2 = 0.9740)

ASK22 Biomass (gL-1) = 0.4918 × Abs680 ̶ 0.0221 (R2 = 0.9986)

ASK25 Biomass (gL-1) = 0.4217 × Abs680 ̶ 0.0462 (R2 = 0.9864)

ASK27 Biomass (gL-1) = 0.4536 × Abs680 ̶ 0.0005 ( R2 = 0.9954)

Table S3

Summary list of isolated potential strains, source location, tentative taxonomic assignment

and 18S rRNA sequence similarity with BLAST taxa.

Strain IDSource

locationTaxonomic assignment Closest BLAST match with similarity

Sequence

size

ASK 01

25°30'12.0"N

81°52'02.7"E

Desmodesmus sp. (MF

939404)Desmodesmus armatus AB917135 (96%)

693

ASK 14 Chlorella sp. (MF 801313) Chlorella vulgaris KJ676110 (98%) 911

ASK 16 Scenedesmus sp. (MK209709) Scenedesmus sp CPC3 KF537681 (99%) 829

ASK 22 25°38'26.0"

N,

81°83'80.0" E

Scenedesmus sp. (MF 945631) Scenedesmus abundans X73995 (99%) 1709

ASK 25 Chlorella sp. (MF 801361) Chlorella sorokiniana KT852969 (99%) 823

ASK 27 Chlorella sp. (MF 927577) Chlorella sorokiniana KU948990 (99%) 821

Fig. S1 Microscopy of selected potential microalgae isolates (A) Desmodesmus sp. ASK01,

(B) Chlorella sp. ASK14, (C) Scenedesmus sp. ASK16, (D) Scenedesmus sp. ASK22, (E)

Chlorella sp. ASK25, (F) Chlorella sp. ASK27.

Fig. S2 FTIR spectrum of (A) dried biomass of ASK22, (B) lipid extracted biomass (LEMB)

of ASK22, (C) crude oil extracted from ASK22, (D) soybean oil and (E-F) TG and DTG of

the Scenedesmus sp. ASK22 biomass (MB) and lipid extracted biomass (LEMB) in the inert

(nitrogen) and oxidative (air) atmosphere.