Ming Chen

Objective: Long-term culture with high serum concentrations increases culture costs, accelerates cell senescence, and limits industrial-scale applications. Serum reduction adaptation is an effective solution for addressing these issues. In this study, we aimed to investigate the effects of two low-serum adaptation strategies on the growth and lactoferrin expression in stably LTF-overexpressing bovine mammary alveolar cells transfected with large T antigen (MAC-T). Methods: Two adaptation protocols of serum concentration reduction and dual-medium gradient adaptation were implemented to progressively reduce serum dependence in MAC-T cells. Results: Under both adaptation modes, cells maintained stable proliferation at 1% fetal bovine serum (FBS). At the 0.5% FBS (serum reduction) and 10% basal control medium, equivalent to 1% FBS in dual-medium adaptation stages, adding 10% knockout™ serum replacement and 1% insulin-transferrin-selenium 5 increased the specific growth rate by 30%, enabling robust cell growth under low-serum conditions. Conclusion: Across all the low-serum conditions, cells retained typical epithelial morphology (cytokeratin 18-positive) while exhibiting consistent LTF mRNA transcription levels, protein expression, and secretory output. These findings validate the scalable low-serum culturing of MAC-T cells and in vitro LTF synthesis.

Flavor, as one of the primary factors that attracts consumers, has always been a crucial indicator for evaluating the quality of food. From processing to final consumption, the conditions that affect consumers’ perception of the aroma of aquatic products can be divided into three stages: aroma formation, release, and signal transmission. Currently, there are few reviews on the formation, release, and perception of aroma in aquatic products, which has affected the product development of aquatic products. This review summarizes aroma formation pathways, the effects of processing methods, characteristic volatile compounds, various identification techniques, aroma-release influencing factors, and the aroma perception mechanisms of aquatic products. The Maillard reaction and lipid oxidation are the main pathways for the formation of aromas in aquatic products. The extraction, identification, and quantitative analysis of volatile compounds reveal the odor changes in aquatic products. The composition of aquatic products and oral processing mainly influence the release of odorants. The characteristic odorants perceived from the nasal cavity should be given more attention. Moreover, the relationship between various olfactory receptors (ORs) and the composition of multiple aromatic compounds remains to be understood. It is necessary to clarify the relationship between nasal cavity metabolism and odor perception, reveal the binding and activation mode of ORs and odor molecules, and establish an accurate aroma prediction model.

Docosahexaenoic acid (DHA), one of the most important ω-3 long-chain polyunsaturated fatty acids, has attracted great attention in recent years because of its significant health benefits for human beings. Traditionally, DHA is obtained from marine fish oil, but this approach depends on marine fishing and has suffered a dramatic fall in the past few years due to overfishing and climate change, which cannot meet the increasing market demand. Microbial DHA production by oleaginous microorganisms has become the current research hotspot. Schizochytrium sp., a heterotrophic thraustochytrid, has become one of the most promising DHA producers because of its safety, fast growth and high DHA content. However, industrial DHA production by Schizochytrium sp. is severely hindered by the high production cost. Many regulation strategies have been developed to enhance DHA production through fermentation optimization and metabolic regulation. In this review, recent advances in metabolic regulation for enhancing DHA production by Schizochytrium sp. are reviewed, from the aspects of key lipogenic enzymes, precursors, transcription factors, lipid peroxidation, transport of non-esterified DHA and stress environment. Graphical abstract

Direct difunctionalization of unsaturated hydrocarbons with organosilicon reagents for the construction of synthetically valuable silicon functionalized compounds is discussed.

From both economic and environmental perspectives, ethylene glycol, the principal constituent in the degradation of PET, emerges as an optimal feedstock for microbial cell factories. Traditional methods for constructing Escherichia coli chassis cells capable of utilizing ethylene glycol as a non-sugar feedstock typically involve overexpressing the genes fucO and aldA. However, these approaches have not succeeded in enabling the exclusive use of ethylene glycol as the sole source of carbon and energy for growth. Through ultraviolet radiation-induced mutagenesis and subsequent laboratory adaptive evolution, an EG02 strain emerged from E. coli MG1655 capable of utilizing ethylene glycol as its sole carbon and energy source, demonstrating an uptake rate of 8.1 ± 1.3 mmol/gDW h. Comparative transcriptome analysis guided reverse metabolic engineering, successfully enabling four wild-type E. coli strains to metabolize ethylene glycol exclusively. This was achieved through overexpression of the gcl, hyi, glxR, and glxK genes. Notably, the engineered E. coli chassis cells efficiently metabolized the 87 mM ethylene glycol found in PET enzymatic degradation products following 72 h of fermentation. This work presents a practical solution for recycling ethylene glycol from PET waste degradation products, demonstrating that simply adding M9 salts can effectively convert them into viable raw materials for E. coli cell factories. Our findings also emphasize the significant roles of genes associated with the glycolate and glyoxylate degradation I pathway in the metabolic utilization of ethylene glycol, an aspect frequently overlooked in previous research.