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Acetate Production from Cafeteria Wastes and Corn Stover Using a Thermophilic Anaerobic Consortium: A Prelude Study for the Use of Acetate for the Production of Value-Added Products.

ABSTRACT: Efficient and sustainable biochemical production using low-cost waste assumes considerable industrial and ecological importance. Solid organic wastes (SOWs) are inexpensive, abundantly available resources and their bioconversion to volatile fatty acids, especially acetate, aids in relieving the requirements of pure sugars for microbial biochemical productions in industries. Acetate production from SOW that utilizes the organic carbon of these wastes is used as an efficient solid waste reduction strategy if the environmental factors are optimized. This study screens and optimizes influential factors (physical and chemical) for acetate production by a thermophilic acetogenic consortium using two SOWs-cafeteria wastes and corn stover. The screening experiment revealed significant effects of temperature, bromoethane sulfonate, and shaking on acetate production. Temperature, medium pH, and C:N ratio were further optimized using statistical optimization with response surface methodology. The maximum acetate concentration of 8061 mg L-1 (>200% improvement) was achieved at temperature, pH, and C:N ratio of 60 °C, 6, 25, respectively, and acetate accounted for more than 85% of metabolites. This study also demonstrated the feasibility of using acetate-rich fermentate (obtained from SOWs) as a substrate for the growth of industrially relevant yeast Yarrowia lipolytica, which can convert acetate into higher-value biochemicals.

SUBMITTER: David A

PROVIDER: S-EPMC7143096 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The production of value-added chemicals alongside biofuels from lignocellulosic hydrolysates is critical for developing economically viable biorefineries. Here, the production of propionic acid (PA), a potential building block for C3-based chemicals, from corn stover hydrolysate is investigated using the native PA-producing bacterium Propionibacterium acidipropionici.A wide range of culture conditions and process parameters were examined and experimentally optimized to maximize titer, rate, and yield of PA. The effect of gas sparging during fermentation was first examined, and N2 was found to exhibit improved performance over CO2. Subsequently, the effects of different hydrolysate concentrations, nitrogen sources, and neutralization agents were investigated. One of the best combinations found during batch experiments used yeast extract (YE) as the primary nitrogen source and NH4OH for pH control. This combination enabled PA titers of 30.8 g/L with a productivity of 0.40 g/L h from 76.8 g/L biomass sugars, while successfully minimizing lactic acid production. Due to the economic significance of downstream separations, increasing titers using fed-batch fermentation was examined by changing both feeding media and strategy. Continuous feeding of hydrolysate was found to be superior to pulsed feeding and combined with high YE concentrations increased PA titers to 62.7 g/L and improved the simultaneous utilization of different biomass sugars. Additionally, applying high YE supplementation maintains the lactic acid concentration below 4 g/L for the duration of the fermentation. Finally, with the aim of increasing productivity, high cell density fed-batch fermentations were conducted. PA titers increased to 64.7 g/L with a productivity of 2.35 g/L h for the batch stage and 0.77 g/L h for the overall process.These results highlight the importance of media and fermentation strategy to improve PA production. Overall, this work demonstrates the feasibility of producing PA from corn stover hydrolysate.

Project description:Introductionβ-Glucosidase serves as the pivotal rate-limiting enzyme in the cellulose degradation process, facilitating the hydrolysis of cellobiose and cellooligosaccharides into glucose. However, the widespread application of numerous β-glucosidases is hindered by their limited thermostability and low glucose tolerance, particularly in elevated-temperature and high-glucose environments.MethodsThis study presents an analysis of a β-glucosidase gene belonging to the GH1 family, denoted lqbg8, which was isolated from the metagenomic repository of Hehua hot spring located in Tengchong, China. Subsequently, the gene was cloned and heterologously expressed in Escherichia coli BL21(DE3). Post expression, the recombinant β-glucosidase (LQBG8) underwent purification through a Ni affinity chromatography column, thereby enabling the in-depth exploration of its enzymatic properties.ResultsLQBG8 had an optimal temperature of 70°C and an optimum pH of 5.6. LQBG8 retained 100 and 70% of its maximum activity after 2-h incubation periods at 65°C and 70°C, respectively. Moreover, even following exposure to pH ranges of 3.0-10.0 for 24 h, LQBG8 retained approximately 80% of its initial activity. Notably, the enzymatic prowess of LQBG8 remained substantial at glucose concentrations of up to 3 M, with a retention of over 60% relative activity. The kinetic parameters of LQBG8 were characterized using cellobiose as substrate, with Km and Vmax values of 28 ± 1.9 mg/mL and 55 ± 3.2 μmol/min/mg, respectively. Furthermore, the introduction of LQBG8 (at a concentration of 0.03 mg/mL) into a conventional cellulase reaction system led to an impressive 43.7% augmentation in glucose yield from corn stover over a 24-h period. Molecular dynamics simulations offered valuable insights into LQBG8's thermophilic nature, attributing its robust stability to reduced fluctuations, conformational changes, and heightened structural rigidity in comparison to mesophilic β-glucosidases.DiscussionIn summation, its thermophilic, thermostable, and glucose-tolerant attributes, render LQBG8 ripe for potential applications across diverse domains encompassing food, feed, and the production of lignocellulosic ethanol.

Dataset Information

Acetate Production from Cafeteria Wastes and Corn Stover Using a Thermophilic Anaerobic Consortium: A Prelude Study for the Use of Acetate for the Production of Value-Added Products.

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