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Metabolic Engineering of De Novo Pathway for the Production of 2'-Fucosyllactose 2'-Fucosyllactose (2'-FL), one of the most abundant oligosaccharides in human milk, has gained increased attention owing to its nutraceutical and pharmaceutical potential. However, limited availability and high-cost of preparation have limited its widespread application and in-depth investigation of its potential functions. Here, a modular pathway engineering was implemented to construct an Escherichia coli strain to improve the biosynthesis titer of 2'-FL. Before overexpression of manB, manC, gmd, wcaG, and heterologous expression of futC, genes wcaJ and lacZ encoding UDP-glucose lipid carrier transferase and β-galactosidase, respectively, were inactivated from E. coli BL21 (DE3) with the CRISPR-Cas9 system, which inhibited the production of 2'-FL. The results showed that final shake flask culture yielded a 3-fold increase in 2'-FL 8&nbsp;gL) from the engineered strain EL7.<br /><br /> Fed- [https://tupalo.com/en/users/6290203 Order immediately] were optimized in a 3-L bioreactor. The highest titer of 2'-FL substrate conversion of8&nbsp;gg lactose.Metabolic engineering strategies of de novo pathway for enhancing [https://able2know.org/user/denimgate5/ ] [https://community.windy.com/user/roofcross9 2'-fucosyllactose] synthesis in Escherichia coli'-Fucosyllactose (2'-FL), one of the most abundant human milk oligosaccharides health benefits for newborns. However, limited availability and high-cost preparation have restricted its extensive use and intensive research on its potential functions. In this work, a powerful Escherichia coli cell factory was developed to ulteriorly increase 2'-FL production. Initially, a modular pathway engineering was strengthened to balance the synthesis pathway through different plasmid combinations with a resulting maximum 2'-FL titre of 15&nbsp;g&nbsp;l-1 . To further facilitate the metabolic flux from GDP-l-fucose towards 2'-FL, the CRISPR-Cas9 system was utilized to inactivate the genes including lacZ and wcaJ, increasing the titre by 69-fold.<br /><br /> Notably, the co-introduction of NADPH and GTP regeneration pathways was confirmed to be more conducive to 2'-FL formation, achieving a 2'-FL titre of 24&nbsp;g&nbsp;l-1 . Moreover, comparisons of various exogenous α1,2-fucosyltransferase candidates revealed that futC from Helicobacter pylori generated the highest titre of 2'-FL. Finally, the viability of scaled-up production of 2'-FL was evidenced in a 3&nbsp;l bioreactor with a Microbiology and John Wiley &amp; Sons Ltd.Conflict of interest statement The authors declare no competing financial 3'-Sialyllactose alleviates bone loss by regulating bone homeostasis.Baek A(#)(1)(2), Baek D(#)(1)(2), Cho Y(3), Jo S(2)(4), Kim J(2)(5), Hong Y(6), Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Republic Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Republic Osteoporosis is a common skeletal disease that results in an increased risk of fractures. However, there is no definitive cure, warranting the development of potential therapeutic agents. 3'-Sialyllactose (3'-SL) in human milk regulates many biological functions.<br /><br /> However, its effect on bone metabolism remains unknown. This study aimed to investigate the molecular mechanisms underlying the effect of 3'-SL on bone homeostasis. Treatment of human bone marrow stromal cells (hBMSCs) with 3'-SL enhanced osteogenic differentiation and inhibited adipogenic differentiation of hBMSCs. RNA sequencing showed that 3'-SL enhanced laminin subunit gamma-2 expression and promoted osteogenic differentiation via the phosphatidylinositol 3‑kinaseprotein kinase B signaling pathway. Furthermore, 3'-SL inhibited the receptor activator of nuclear factor κB ligand-induced osteoclast differentiation of bone marrow-derived macrophages through the nuclear factor κB and mitogen‑activated protein kinase signaling pathway, ameliorated osteoporosis in ovariectomized mice, and positively regulated bone remodeling. Our findings suggest 3'-SL as a potential drug for Conflict of interest statement The authors declare no competing interests.[An immunoenzyme system for determining antibodies to Streptococcus pneumoniae polysaccharides in biological fluids].<br /><br />Padiukov LN, Ulanova MA, Kuznetsova EM, Ksenofontova MK, Kulak IuV, Sharapova The enzyme immunoassay (EIA) system for the determination of antibodies to capsular polysaccharides of pneumococci, serotypes 1, 3, 6B, 8, 9N, 15F, 23F, and C-polysaccharide has been developed on the basis of poly-L-lysin-modified antigens.