###### Abstract  
Overcoming obstacles to commercialization of algal-based processes for biofuels and co-products requires not just piecemeal incremental improvements, but rather a comprehensive and fundamental re-consideration starting with the selected algae and its associated cultivation, harvesting, biomass conversion, and refinement. A novel two-stage process designed to address challenges of mass outdoor microalgal cultivation for biofuels and co-products was previously demonstrated using an oleaginous, haloalkaline-tolerant, and multi-trophic green _Chlorella vulgaris_. ALP2 from a soda lake. This involved cultivating the microalgae in a fermenter heterotrophically or photobieroactor mixotrophically (first-stage) to rapidly obtain high cell densities and inoculate an open-pond phototrophic culture (second-stage) featuring high levels of NaHCO\({}_{3}\) pH, and salinity. An improved two-stage cultivation that instead sustainably used as more cheap and sustainable inputs the organic carbon, nitrogen, and phosphorous from fractionation of waste was here demonstrated in a small-scale biorefinery process. The first cultivation stage consisted of two simultaneous batch flask cultures featuring (1) mixotrophic cell productivity of 7.25 \(\times\) 10\({}^{7}\) cells mL\({}^{-1}\) day\({}^{-1}\) on BG-11\({}_{0}\) medium supplemented with 1.587 g L\({}^{-1}\) urea and an enzymatic hydrolysate of pre-treated (torrefaction + grinding + ozonolysis + soaking ammonia) wheat-straw that corresponded to 10 g L\({}^{-1}\) glucose, and (2) mixotrophic cell productivity of 2.25 \(\times\) 10\({}^{7}\) cells mL\({}^{-1}\) day\({}^{-1}\) on BG-11\({}_{0}\) medium supplemented with 1.587 g L\({}^{-1}\) urea and a purified and de-toxified condensate of pre-treated (torrefaction + grinding) wheat straw that corresponded to 0.350 g L\({}^{-1}\) of potassium acetate. The second cultivation stage featured \({}^{1}\)H NMR-determined phototrophic lipid productivity of 0.045 g tricacylglycereptides (TAG) L\({}^{-1}\) day\({}^{-1}\) on BG-11\({}_{0}\) medium supplemented with 16.8 g L\({}^{-1}\) NaHCO\({}_{3}\) and fed batch-added 22% (\(\
u/v\)) anaerobically digested food waste effluent at HCI-mediated pH 9.  
biorefinery; algae; anaerobic digestion; torrefaction; enzyme hydrolysis; food and lignocellulosic waste +
Footnote †: journal: _Sciences_  
_Applied_  
_Acticle_  
Biorefinery Processing of Waste to Supply Cost-Effective and Sustainable Inputs for Two-Stage Microalgal Cultivation  
**[PERSON]  
1 Department of Biological Systems Engineering, Washington State University, Pullman, WA 99164, USA; [EMAIL_ADDRESS] (M.B. ); [EMAIL_ADDRESS] (A.G.);[EMAIL_ADDRESS] (M.R.P.-S.); [EMAIL_ADDRESS] (L.Y. ); [EMAIL_ADDRESS] (M.G.-P.)  
2 Department of Applied Chemistry and Systems of Production, Universidad de Cuenca, Cuenca 010158, Ecuador 2  
Centre for NMR Spectroscopy, Washington State University, Pullman, WA 99164, USA; [EMAIL_ADDRESS] (W.H. ); [EMAIL_ADDRESS] (C.L.H.) 4 College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA; [EMAIL_ADDRESS]  
5 Institute of Biological Chemistry, Washington State University, Pullman, WA 99164, USA; [EMAIL_ADDRESS]  
* Correspondence: [EMAIL_ADDRESS] (P.C.W. ); [EMAIL_ADDRESS] (S.C.); Tel. : +1-509-335-3743 (S.C.); Fax: +1-509-335-2722 (S.C.)  
**Featured Application: This described technology can be applied for cost-effective manufacturing of food, feed, fuel, therapeutics, and bioplastics via algal cultivation. **  
**Citation:** [PERSON]; [PERSON]; [PERSON]; [PERSON]; [PERSON]; [PERSON]; [PERSON]; [PERSON]; [PERSON]; [PERSON] [PERSON]; et al. Biofinery Processing of Waste to Supply Cost-Effective and Sustainable Inputs for Two-Stage Microalgal Cultivation. **Academic Editor: [PERSON]**  
Received: 21 November 2021  
Accepted: 24 January 2022  
Published: 29 January 2022  
**Publisher:**  
inorganic carbon, and water, exposure to environmental factors, and limited available land acreage for inoculum and subsequent large-scale cultures.