In this study, the feasibility of promoting the lactic acid (LA) fermentation of food waste (FW) with iron tailings (ITs) addition was explored. The best LA yield was 0.91 g LA/g total sugar when 1 % ITs were added into the system. The mechanisms for promoting LA production were acidification alleviation effects and reduction equivalent supply of ITs. Furthermore, the addition of ITs promoted carbohydrate hydrolysis, and the carbohydrates digestibility reached 88.85 % in the 1 % ITs group. The ITs also affected the microbial communities, Lactococcus gradually replaced Streptococcus as the dominant genus, and results suggested that Lactococcus had a positive correlation with LA production and carbohydrate digestibility. Finally, the complex LAB in FW had significant effects on heavy metal removal from ITs, and the removal efficiency Cr, As, Pb, Cd, and Hg can reach 50.84 %, 26.72 %, 59.65 %, 49.75 % and 78.87 % in the 1 % ITs group, respectively.

In recent years, with the booming of the edible mushroom industry, chitin production has become increasingly dependent on fungi and other non-traditional sources. Fungal chitin has advantages including superior performance, simpler separation processes, abundant raw materials, and the absence of shellfish allergens. As a kind of edible mushroom, flammulina velutipes (F. velutipes) also has the advantages of wide source and large annual yield. This provided the possibility for the extraction of chitin. Here, a procedure to extract chitin from F. velutipes waste be presented. This method comprises low-concentration acid pretreatment coupled with consolidated bioprocessing with Aspergillus niger. Characterization by SEM, FTIR, XRD, NMR, and TGA confirmed that the extracted chitin was β-chitin. To achieve optimal fermentation of F. velutipes waste (80 g/L), ammonium sulfate and glucose were selected as nitrogen and carbon sources (5 g/L), with a fermentation time of 5 days. The extracted chitin could be further deacetylated and purified to obtain high-purity chitosan (99.2 % ± 1.07 %). This chitosan exhibited a wide degree of deacetylation (50.0 % ± 1.33 % - 92.1 % ± 0.97 %) and a molecular weight distribution of 92-192 kDa. Notably, the yield of chitosan extracted in this study was increased by 56.3 % ± 0.47 % compared to the traditional chemical extraction method.

Manipulation of the lactate metabolism is an efficient way for cancer treatment given its involvement in cancer development, metastasis, and immune escape. However, most of the inhibitors of lactate transport carriers suffer from poor specificity. Herein, we use the CRISPR/Cas9 system to precisely downregulate the monocarboxylate carrier 1 (MCT1) expression. To avoid the self-repairing during the gene editing process, a dual-Cas9 ribonucleoproteins (duRNPs) system is generated using the biological fermentation method and delivered into cells by the zeolitic imidazolate framework-8 (ZIF-8) nanoparticles, enabling precise removal of a specific DNA fragment from the genome. For efficient cancer therapy, a specific glucose transporter 1 inhibitor (BAY-876) is co-delivered with the duRNPs, forming BAY/duRNPs@ZIF-8 nanoparticle. ZIF-8 nanoparticles can deliver the duRNPs into cells within 1 h, which efficiently downregulates the MCT1 expression, and prohibits lactate influx. Through simultaneous inhibition of the lactate and glucose influx, BAY/duRNPs@ZIF-8 prohibits ATP generation, arrests cell cycle, inhibits cell proliferation, and finally induces cellular apoptosis both in vitro and in vivo. Consequently, we demonstrate that the biologically produced duRNPs delivered into cells by the nonviral ZIF-8 carrier have expanded the CRISPR/Cas gene editing toolbox and elevated the gene editing efficiency, which will promote biological studies and clinical applications. STATEMENT OF SIGNIFICANCE: The CRISPR/Cas9 system, widely used as an efficient gene editing tool, faces a challenge due to cells\' ability to self-repair. To address this issue, a strategy involving dual-cutting of the genome DNA has been designed and implemented. This strategy utilizes biologically produced dual-ribonucleoproteins delivered by a metal-organic framework. The effectiveness of this dual-cut CRISPR-Cas9 system has been demonstrated through a therapeutic approach targeting the simultaneous inhibition of lactate and glucose influx in cancer cells. The utilization of the dual-cut gene editing strategy has provided valuable insights into gene editing and expanded the toolbox of the CRISPR/Cas-based gene editing system. It has the potential to enable more efficient and precise manipulation of specific protein expression in the future.

This study aimed to screen an appropriate starter and determine the optimal fermentation process parameters to optimize the fermentation process of nutrient components and bioactive contents in pharyngitis tablet residue. This study included two experiments. In experiment I, single-factor experimental design was used to study the effects of different biological starters (cellulase preparation; Lactobacillus Plantarum and Bacillus subtilis preparation; mixture of cellulase + Lactobacillus Plantarum and Bacillus subtilis) on the nutritional values and bioactive ingredient contents in pharyngitis tablet residue. In experiment II, orthogonal design experiment was adopted to study the effects of initial water content (45, 50, and 55%), fermentation temperature (35, 37, and 40°C), and fermentation time (24, 48, and 72 h) on the changes in nutrient components, biologically activity contents, and toxin contents of the residue after optimal fermentation agent treatment. Cellulase preparation was found to be the optimal starter. The optimal fermentation conditions were: initial water content, 55%; fermentation temperature, 37°C; and fermentation time, 72 h. The contents of aflatoxin B1 and vomit toxin were in line with Chinese feed hygiene production standards. The fermentation quality of pharyngitis tablet residue can be improved by using the optimal starter and fermentation conditions.

The combination of 3D printing and synthetic biology is a sustainable strategy to fabricate biological objects and systems that behave in a preprogrammed manner. Many microorganisms have been genetically engineered as cell factories for the biosynthesis of chemicals using fermentation, and 3D printing of living materials using these cells could lead to a new paradigm for biomanufacturing.

Low concentration alcohols produced by state-of-the-art biological fermentation restrict subsequent purification processes for chemical, pharmaceutical, biofuel, and other applications. Herein, a rarely reported cucurbituril[n] (n = 6, 8) is employed to pattern the thin-film composite membranes with controllable and quantifiable nanostrand structures through a host-guest strategy. The resulting nanofiltration membrane with such morphology is the first report that exhibits excellent separation performance for isopropyl alcohol (IPA) and water, condensing the initial 0.5 wt % IPA aqueous solution to 9.0 wt %. This not only provides a novel strategy for patterning nanostructural morphology but also makes nanofiltration membranes promising for alcohol condensation in the biological fermentation industry that may reduce energy consumption and postprocessing costs.