Browsing by Author "Misra, Ashish"

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    Development of hypertolerant strain of Yarrowia lipolytica accumulating succinic acid using high levels of acetate
    (American Chemical Society, 2022-08-09) Narisetty, Vivek; Prabhu, Ashish A.; Bommareddy, Rajesh Reddy; Cox, Rylan; Agrawal, Deepti; Misra, Ashish; Ali Haider, M.; Bhatnagar, Amit; Pandey, Ashok; Kumar, Vinod
    Acetate is emerging as a promising feedstock for biorefineries as it can serve as an alternate carbon source for microbial cell factories. In this study, we expressed acetyl-CoA synthase in Yarrowia lipolytica PSA02004PP, and the recombinant strain grew on acetate as the sole carbon source and accumulated succinic acid or succinate (SA). Unlike traditional feedstocks, acetate is a toxic substrate for microorganisms; therefore, the recombinant strain was further subjected to adaptive laboratory evolution to alleviate toxicity and improve tolerance against acetate. At high acetate concentrations, the adapted strain Y. lipolytica ACS 5.0 grew rapidly and accumulated lipids and SA. Bioreactor cultivation of ACS 5.0 with 22.5 g/L acetate in a batch mode resulted in a maximum cell OD600 of 9.2, with lipid and SA accumulation being 0.84 and 5.1 g/L, respectively. However, its fed-batch cultivation yielded a cell OD600 of 23.5, SA titer of 6.5 g/L, and lipid production of 1.5 g/L with an acetate uptake rate of 0.2 g/L h, about 2.86 times higher than the parent strain. Cofermentation of acetate and glucose significantly enhanced the SA titer and lipid accumulation to 12.2 and 1.8 g/L, respectively, with marginal increment in cell growth (OD600: 26.7). Furthermore, metabolic flux analysis has drawn insights into utilizing acetate for the production of metabolites that are downstream to acetyl-CoA. To the best of our knowledge, this is the first report on SA production from acetate by Y. lipolytica and demonstrates a path for direct valorization of sugar-rich biomass hydrolysates with elevated acetate levels to SA.
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    Fermentative production of 2,3-Butanediol using bread waste – A green approach for sustainable management of food waste
    (Elsevier, 2022-06-01) Narisetty, Vivek; Zhang, Le; Zhang, Jingxin; Lin, Carol Sze Ki; Tong, Yen Wah; Show, Pau Loke; Bhatia, Shashi Kant; Misra, Ashish; Kumar, Vinod
    Bread is Europe’s most wasted food, and the second most wasted food after potatoes in UK. Bread waste (BW) is a clean source of high-quality fermentable sugars. In this study, the potential of Enterobacter ludwigii to accumulate 2,3-butanediol (BDO) from BW was evaluated. Initially, the optimal inoculum size and yeast extract concentration were determined, followed by extraction of sugars from BW using acid and enzymatic hydrolysis. A glucose yield of 330–530 g/kg BW was obtained, and the sugars released were utilised for BDO production by E. ludwigii. The fed-batch cultivation using pure glucose and glucose rich hydrolysates from acid and enzymatic hydrolysis resulted in BDO titres of 144.5, 135.4, and 138.8 g/L, after 96 h, with yield of 0.47, 0.42 and 0.48 g/g yield, respectively. The innovation of the work is valorisation of BW to BDO with a circular biorefining approach and thus, reducing BW disposal and associated environmental burden.