Sustainable succinic acid production from lignocellulosic hydrolysates by engineered strains of Yarrowia lipolytica at low pH.

Succinic acid (SA) is a valuable C4 platform chemical with diverse applications. Lignocellulosic biomass represents an abundant and renewable carbon resource for microbial production of SA. However, the presence of toxic compounds in pretreated lignocellulosic hydrolysates poses challenges to cell metabolism, leading to inefficient SA production. Here, engineered Yarrowia lipolytica Hi-SA2 was shown to utilize glucose and xylose from corncob hydrolysate to produce 32.6 g/L SA in shaking flasks. The high concentration of undetoxified hydrolysates significantly inhibited yeast growth and SA biosynthesis, with furfural identified as the key inhibitor. Through overexpressing glutathione synthetase encoding gene YlGsh2, the tolerance of engineered strain to furfural and toxic hydrolysate was significantly improved. In a 5-L bioreactor, Hi-SA2-YlGsh2 strain produced 45.34 g/L SA within 32 h, with a final pH of 3.28. This study provides a sustainable process for bio-based SA production, highlighting the efficient SA synthesis from lignocellulosic biomass through low pH fermentation.

Nanofiber Composite Microchannel-Containing Injectable Hydrogels for Cartilage Tissue Regeneration.

Articular cartilage tissue is incapable of self-repair and therapies for cartilage defects are still lacking. Injectable hydrogels have drawn much attention in the field of cartilage regeneration. Herein, the novel design of nanofiber composite microchannel-containing hydrogels inspired by the tunnel-piled structure of subway tunnels is proposed. Based on the aldehydized polyethylene glycol/carboxymethyl chitosan (APA/CMCS) hydrogels, thermosensitive gelatin microrods (GMs) are used as a pore-forming agent, and coaxial electrospinning polylactic acid/gelatin fibers (PGFs) loaded with kartogenin (KGN) are used as a reinforcing agent and a drug delivery system to construct the nanofiber composite microchannel-containing injectable hydrogels (APA/CMCS/KGN@PGF/GM hydrogels). The in situ formation, micromorphology and porosity, swelling and degradation, mechanical properties, self-healing behavior, as well as drug release of the nanofiber composite microchannel-containing hydrogels are investigated. The hydrogel exhibits good self-healing ability, and the introduction of PGF nanofibers can significantly improve the mechanical properties. The drug delivery system can realize sustained release of KGN to match the process of cartilage repair. The microchannel structure effectively promotes bone marrow mesenchymal stem cell (BMSC) proliferation and ingrowth within the hydrogels. In vitro and animal experiments indicate that the APA/CMCS/KGN@PGF/GM hydrogels can enhance the chondrogenesis of BMSCs and promote neocartilage formation in the rabbit cartilage defect model.

Screening lactic acid bacteria and yeast strains for soybean paste fermentation in northeast of China.

Soybean paste was a traditional fermented product in northeast China, mainly fermented by molds, yeast, Bacillus, and lactic acid bacteria. In this study, the safety and fermentation ability of lactic acid bacteria and yeast strains isolated from traditional soybean paste in northeast China were evaluated, and the dynamic changes of biogenic amines, aflatoxin, total acids, amino acid nitrogen, and volatile compounds were investigated during the fermentation of the traditional soybean paste. Among the tested strains, Lactiplantibacillus plantarum DPUL-J8 could decompose putrescine by 100%, and no biogenic amine was produced by Pichia kudriavzevii DPUY-J8. Lactiplantibacillus plantarum DPUL-J8 and P. kudriavzevii DPUY-J8 with strong biogenic amine degrading capacities were inoculated into the soybean paste. After 30 days of fermentation, the content of biogenic amines and aflatoxin in the fermented soybean paste declined by more than 60% and 50%, respectively. At the same time, compared with the control group without inoculation, the contents of total acid (1.29 ± 0.05 g/100 g), amino acid nitrogen (0.82 ± 0.01 g/100 g), and volatile compounds in soybean paste fermented by L. plantarum DPUL-J8 and P. kudriavzevii DPUY-J8 were significantly increased, which had a good flavor. These results indicated that the use of L. plantarum DPUL-J8 and P. kudriavzevii DPUY-J8 as starter cultures for soybean paste might be a good strategy to improve the safety and flavor of traditional Chinese soybean paste.

Safety evaluation and application of lactic acid bacteria and yeast strains isolated from Sichuan broad bean paste.

Broad bean paste is one of the most popular characteristic traditional fermented bean products in China, which is prepared by mixed fermentation of a variety of microorganisms, among which lactic acid bacteria and yeast played an important role in the improvement of the fermented broad bean paste quality. However, the traditional open-air fermentation of broad bean paste brought some risks of harmful microorganisms. In this study, the safety and fermentation ability of lactic acid bacteria and yeast strains isolated from traditional broad bean paste was evaluated. The results showed that the protease activity of the strain Lactobacillus plantarum DPUL-J5 (366.73 ± 9.00 U/L) and Pichia kudriavzevii DPUY-J5 (237.18 ± 10.93 U/L) were the highest. Both strains produced little biogenic amines, and did not exhibit α-hemolytic activity or antibiotic resistance for some of the antibiotics most used in human medicine. Furthermore, the broad bean paste fermentation involving DPUL-J5 and DPUY-J5 was beneficial for accumulating higher total acid (1.69 ± 0.01 g/100 g), amino-acid nitrogen (0.85 ± 0.03 g/100 g), and more volatile flavor compounds, meanwhile, reducing the levels of biogenic amines and aflatoxin B1. Therefore, this study provided a new strategy to improve the safety and quality of traditional broad bean paste.

Boron doping positively enhances the catalytic activity of carbon materials for the removal of bisphenol A.

Boron-doped carbon materials (BCs), low-cost and environmentally friendly carbocatalysts, were prepared for the activation of persulfate (PS) for the removal of bisphenol A (BPA). Compared with B-free carbon materials (Cs), the adsorption and catalytic activity were significantly enhanced by the boron modification. Fast and efficient removal of BPA was achieved using the BCs/PS system. The BPA removal rate constant increased linearly with the adsorption capacity of BCs. Electron paramagnetic resonance (EPR) spectroscopy and radical quenching experiments indicated that the degradation mechanisms in the BCs/PS system were different from conventional radical-based oxidation pathways. On the contrary, nonradical pathways were demonstrated to dominate the oxidation processes in the removal of BPA using the BCs/PS system. Herein, a mechanism is proposed where PS is activated by the carbon material to form a reactive electron-deficient carbocatalyst ([BCs]*) complex with a high redox potential, driving a nonradical oxidation pathway to achieve BPA removal. Through experimental investigation and the use of electrochemical techniques (cyclic voltammetry, Tafel corrosion analysis and open circuit voltages), B-doped carbon materials for the activation of PS elevate the potential of the derived nonradical [BCs]* complexes, and then accelerate the BPA removal efficiency via an electron transfer process. Utilizing adsorption and nonradical oxidation processes, the BCs/PS system possesses great potential for the removal of BPA in practical applications such as wastewater treatment.

Enhancing thermal stability and lytic activity of phage lysin PlyAB1 from Acinetobacter baumannii.

With the increasingly serious drug resistance of Acinetobacter baumannii, there is an increasingly urgent need for new antibacterial drugs. Phage lysin PlyAB1 has a bactericidal effect on drug-resistant A. baumannii, which has the potential to replace antibiotics to fight infection caused by A. baumannii. However, its application is limited by its thermal stability and lytic activity. To solve these problems, molecular dynamics (MD) simulations combined with Hotspot wizard 3.0 were used to identify key residue sites affecting thermal stability, and evolutionary analysis combined with multiple sequence alignment was used to identify key residue sites affecting lytic activity. Four single-point variants with significantly increased thermal stability and four single-point variants with significantly lytic activity were obtained, respectively. Furthermore, by superimposing mutations, we obtained three double-point variants, G100Q/K69R, G100R/K69R, and G100K/K69R, with significantly improved thermal stability and improved lytic activity. At 45°C, the lytic activity and half-life of the optimal variant G100Q/K69R were 1.51- and 24-fold higher than those of the wild PlyAB1, respectively. These results deepen our understanding of the structure and function of phage lysin and contribute to the application of phage lysin in antibiotic substitution.

Vitamin D status of pregnant women in Shanghai, China.

OBJECTIVE: The aims of this study were to identify the prevalence of vitamin D insufficiency in pregnant women in Shanghai, China and to determine the correlations between 25-hydroxyvitamin D (25(OH)D) and age, serum lipids, renal function, BMI, gestational weeks, and season of sampling. METHODS: A total of 1695 pregnant women who visited the outpatient clinic for their first prenatal examination were included in the study. RESULTS: Over 90.5% of these women had less than optimal levels of 25(OH)D. Of the 1695 women, 1162 were deficient, 372 were insufficient, only 372 were sufficient, and 5 had possible hypervitaminosis. 25(OH)D levels were negatively correlated with age, total cholesterol, low-density lipoprotein (LDL), gestational age, body weight, and BMI. They were positively correlated with urea levels. Neither parity nor season was a significant factor. CONCLUSIONS: Vitamin D deficiency and insufficiency were common in pregnant Chinese women and routine screening as well as prophylactic measures should be considered.