Browsing by Author "Koutinas, Apostolis"

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    Enhanced 2,3-Butanediol production by mutant Enterobacter ludwigii using Brewers’ spent grain hydrolysate: process optimization for a pragmatic biorefinery loom
    (Elsevier, 2021-06-18) Amraoui, Yassin; Prabhu, Ashish A.; Narisetty, Vivek; Coulon, Frederic; Chandel, Anuj Kumar; Willoughby, Nicholas; Jacob, Samuel; Koutinas, Apostolis; Kumar, Vinod
    2,3-Butanediol (BDO) is a fossil-based versatile bulk chemical with a multitude of applications. BDO can also be synthesized using microbial cell factories harnessing renewable feedstocks. However, the high cost of the substrate via microbial route impedes commercial manufacturing of BDO. Therefore, identification of cheaper substrates could make bio-based BDO production more cost-competitive. Brewers’ spent grain (BSG), a major by-product of breweries, is an inexpensive source of fermentable sugars and proteins. In the present study, we have attempted the bioproduction of BDO by Enterobacter ludwigii using BSG as feedstock. A random E. ludwigii mutant obtained after treatment with ethyl methane sulfonate (EMS) resulted in a BDO titer (9.5 g/L), ~30% higher in comparison to the wild type strain with a yield of 0.48 gBDO/gGlucose approaching the theoretical yield of 0.50 gBDO/gGlucose. The enzymatic hydrolysis of microwave-assisted alkali pretreated BSG was optimized using the statistical Taguchi design. The BSG hydrolysis under optimal conditions (pH: 6.0; temperature: 50 °C; BSG: 10% w/v; enzyme loading: 2% v/v) resulted in a glucose yield of 0.25 gGlucose/gBiomass. The uncontrolled pH was found to be more beneficial for BDO accumulation from BSG hydrolysate in batch bioreactor cultivation as compared with controlled one. The fed-batch cultivation with forced pH fluctuations at an aeration rate of 2.0 vvm resulted in BDO accumulation of 118.5 g/L from glucose-rich BSG hydrolysate with the yield and productivity of 0.43 g/g and 1.65 g/L.h, respectively. To the best of our knowledge, this is the first study on BDO production from BSG.
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    Integrated fermentative production and downstream processing of 2,3-butanediol from sugarcane bagasse-derived xylose by mutant strain of Enterobacter ludwigii
    (American Chemical Society, 2021-07-16) Amraoui, Yassin; Narisetty, Vivek; Coulon, Frederic; Agrawal, Deepti; Chandel, Anuj Kumar; Maina, Sofia; Koutinas, Apostolis; Kumar, Vinod
    In this study, a mutant strain of Enterobacter ludwigii developed in our previous work, was evaluated to utilize pure xylose as the sole carbon and energy source for 2,3-butanediol (BDO) production. Later, this strain was also investigated on detoxified and nondetoxified xylose-rich hydrolysate obtained from hydrothermally pretreated sugarcane bagasse (SCB) for BDO production. Supplementing the fermentation medium with 0.2% w/v yeast extract improved cell growth (31%), BDO titer (43%), and yield (41%) against the synthetic medium devoid of any complex nitrogen source. The fed-batch culture with cyclic control of pH resulted in a BDO production of 71.1 g/L from pure xylose with overall yield and productivity of 0.40 g/g and 0.94 g/L·h, respectively. While BDO titer, yield, and productivity of 63.5 g/L, 0.36 g/g, and 0.84 g/L·h, were acheived with detoxified hydrolysate, respectively. In contrast, 32.7 g/L BDO was produced from nondetoxified hydrolysate with a conversion yield of 0.33 g/g and a productivity of 0.43 g/L·h. BDO accumulated on pure xylose and detoxified SCB hydrolysate was separated by aqueous two-phase system (ATPS) method using (NH4)2SO4 as salting-out agent and isopropanol as an extractant, resulting in the BDO recovery of more than 85%. The results achieved in the current work exemplify a step toward industrial BDO production from cost-effective hemicellulosic hydrolysates by E. ludwigii.
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    Prospects on bio-based 2,3-butanediol and acetoin production: recent progress and advances
    (Elsevier, 2021-06-29) Maina, Sofia; Prabhu, Ashish A.; Vivek, Narisetty; Vlysidis, Anestis; Koutinas, Apostolis; Kumar, Vinod
    The bio-based platform chemicals 2,3-butanediol (BDO) and acetoin have various applications in chemical, cosmetics, food, agriculture, and pharmaceutical industries, whereas the derivatives of BDO could be used as fuel additives, polymer and synthetic rubber production. This review summarizes the novel technological developments in adapting genetic and metabolic engineering strategies for selection and construction of chassis strains for BDO and acetoin production. The valorization of renewable feedstocks and bioprocess development for the upstream and downstream stages of bio-based BDO and acetoin production are discussed. The techno-economic aspects evaluating the viability and industrial potential of bio-based BDO production are presented. The commercialization of bio-based BDO and acetoin production requires the utilization of crude renewable resources, the chassis strains with high fermentation production efficiencies and development of sustainable purification or conversion technologies.