Efficient Production of Triacetic Acid Lactone from Lignocellulose Hydrolysate by Metabolically Engineered Yarrowia lipolytica.

Lignocellulose is a promising renewable feedstock for the bioproduction of high-value biochemicals. The poorly expressed xylose catabolic pathway was the bottleneck in the efficient utilization of the lignocellulose feedstock in yeast. Herein, multiple genetic and process engineering strategies were explored to debottleneck the conversion of xylose to the platform chemical triacetic acid lactone (TAL) in Yarrowia lipolytica. We identified that xylose assimilation generating more cofactor NADPH was favorable for the TAL synthesis. pH control improved the expression of acetyl-CoA carboxylase and generated more precursor malonyl-CoA. Combined with the suppression of the lipid synthesis pathway, 5.03 and 4.18 g/L TAL were produced from pure xylose and xylose-rich wheat straw hydrolysate, respectively. Our work removed the bottleneck of the xylose assimilation pathway and effectively upgraded wheat straw hydrolysate to TAL, which enabled us to build a sustainable oleaginous yeast cell factory to cost-efficiently produce green chemicals from low-cost lignocellulose by Y. lipolytica.

Recent advances and perspectives on production of value-added organic acids through metabolic engineering.

Organic acids are important consumable materials with a wide range of applications in the food, biopolymer and chemical industries. The global consumer organic acids market is estimated to increase to $36.86 billion by 2026. Conventionally, organic acids are produced from the chemical catalysis process with petrochemicals as raw materials, which posts severe environmental concerns and conflicts with our sustainable development goals. Most of the commonly used organic acids can be produced from various organisms. As a state-of-the-art technology, large-scale fermentative production of important organic acids with genetically-modified microbes has become an alternative to the chemical route to meet the market demand. Despite the fact that bio-based organic acid production from renewable cheap feedstock provides a viable solution, low productivity has impeded their industrial-scale application. With our deeper understanding of strain genetics, physiology and the availability of strain engineering tools, new technologies including synthetic biology, various metabolic engineering strategies, omics-based system biology tools, and high throughput screening methods are gradually established to bridge our knowledge gap. And they were further applied to modify the cellular reaction networks of potential microbial hosts and improve the strain performance, which facilitated the commercialization of consumable organic acids. Here we present the recent advances of metabolic engineering strategies to improve the production of important organic acids including fumaric acid, citric acid, itaconic acid, adipic acid, muconic acid, and we also discuss the current challenges and future perspectives on how we can develop a cost-efficient, green and sustainable process to produce these important chemicals from low-cost feedstocks.

Parametric optimization and kinetic study of l-lactic acid production by homologous batch fermentation of Lactobacillus pentosus cells.

Parametric optimization always plays important roles in bioengineering systems to obtain a high product yield under the proper conditions. The parametric conditions of lactic acid production by homologous batch fermentation of Lactobacillus pentosus cells was optimized by the Box-Behnken design. The highest l-lactic acid yield was obtained as 0.836 ± 0.003 g/g glucose with the productivity of 0.906 ± 0.003 g/(L × H) under the optimum conditions of 34.7 °C, pH 6.2, 148 rpm agitation speed, and 9.3 g/L nitrogen source concentration determined by quadratic response surface with high accuracy. The adequate kinetic models of cell growth rate, lactic production rate, and glucose consumption rate were also established to describe the fermentation behavior of L. pentosus cells with the correlation coefficients of 09985, 0.9990, and 0.9989, respectively.

Optimization of immobilization conditions for Lactobacillus pentosus cells.

In this study, the immobilization technology was used to improve the LA yield and shorten the fermentation time. The optimum conditions to immobilize Lactobacillus pentosus ATCC 8041 cell were determined by Taguchi design L16 (45). The immobilized L. pentosus ATCC 8041 cells prepared by 2% sodium alginate (SA) and 6% polyvinyl alcohol (PVA) with the immobilization process by 0.10 M calcium chloride (CaCl2) and 2.5% boric acid (H3BO3) had the best performance of LA yield at the temperature of 35 °C, which is significantly higher than that of L. pentosus ATCC 8041 free cells. These cells maintained the stable and efficient performance in 15 repeated batch fermentation, and they also have excellent mechanical strength to keep from breakage caused by cell growth and agitation.