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Background Main bioactive constituents and pharmacological functions of ripened red ginseng berry (Panax ginseng Meyer) have been frequently reported. Yet, the research gap targeting the beneficial activities of transformed green ginseng berries has not reported elsewhere. Methods Ginsenosides of new green berry cultivar K-1 (GK-1) were identified by HPLC-QTOF/MS. Ginsenosides bioconversion in GK-1 by bgp1 enzyme was confirmed with HPLC and TLC. Then, mechanisms of GK-1 and β-glucosidase (bgp1) biotransformed GK-1 (BGK-1) were determined by Quantitative Reverse Transcription-Polymerase Chain Reaction and Western blot. Results GK-1 possesses highest ginsenosides especially ginsenoside-Re amongst seven ginseng cultivars including (Chunpoong, Huangsuk, Kumpoong, K-1, Honkaejong, Gopoong, and Yunpoong). Ginseng root’s biomass is not affected with the harvest of GK-1 at 3 weeks after flowering period. Then, Re is bio-converted into a promising pharmaceutical effect of Rg2 via bgp1. According to the results of cell assays, BGK-1 shows decrease of tyrosinase and melanin content in α-melanocyte-stimulating hormone challenged-murine melanoma B16 cells. BGK-1 which is comparatively more effective than GK-1 extract shows significant suppression of the nuclear factor (NF)-κB activation and inflammatory target genes, in LPS-stimulated RAW 264.7 cells. Conclusion These results reported effective whitening and anti-inflammatory of BGK-1 as compared to GK-1., Graphical abstract Image 1

Xiao-Jie Mi,1 Xing Yue Xu,1 Han Sol Choi,1 Hoon Kim,1 Ik Hyun Cho,2 Tae-Hoo Yi,1 Yeon-Ju Kim1 1Graduate School of Biotechnology, and College of Life Science, Kyung Hee University, Yongin-si, 17104, Gyeonggi-do, Republic of Korea; 2Department of Science in Korean Medicine and Brain Korea 21 Plus Program, Graduate School, Kyung Hee University, Seoul, 02447, Republic of KoreaCorrespondence: Yeon-Ju Kim; Ik Hyun Cho Tel +82-31-201-5634Fax +82-31-204-8116Email yeonjukim@khu.ac.kr; ihcho@khu.ac.krBackground: Despite great advances in the field of immunotherapy, there is still a need for novel and effective immunostimulants to overcome challenges, such as instability and autoinflammatory toxicity, associated with conventional immunostimulants. Nanotechnology provides the possibility to overcome these challenges. The well-known classical Chinese formula, Hwanglyeonhaedok-tang (HHT) has been widely used to treat immune-related diseases in clinical practice.Methods: We developed novel gold nanoparticles (AuNPs) utilizing one-pot synthesis with the herbal formula-HHT. The optimal conditions for HHT-AuNP biosynthesis were established, and physicochemical properties of the optimized HHT-AuNPs were identified using various spectrometric and microscopic techniques. Bio-TEM analysis revealed that HHT-AuNPs were highly engulfed within RAW264.7 cells without inducing cytotoxicity. The effect of HHT-AuNPs on immunostimulatory activity was evaluated in innate and adaptive immune cells (RAW264.7 macrophages and ICR mice splenocytes) using qRT-PCR, immunoblotting, and ELISA.Results: The HHT-AuNPs remarkably increased the nitric oxide (NO) and immune-related cytokines production by activating the mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signaling pathways in RAW264.7 cells. Furthermore, HHT-AuNPs exerted immunostimulatory effects on mouse splenocytes by priming T/B-cells and macrophages.Discussion: The present study is the first to demonstrate that HHT-AuNPs could be utilized as immunostimulators to activate both innate and adaptive immune systems. These results provide a foundation for the application of traditional Chinese medicinal formulae in the field of nanomedicine.Keywords: HHT, herb formula, green synthesis, AuNPs, immunostimulation

Postbiotics defined as soluble factors (products or metabolic byproducts) that are released after bacterial lysis or secreted by live bacteria, have attracted considerable attention because of their long shelf life, safety, and beneficial effects. In this study, we investigated the immune-enhancing activities of squid jeotgal (a traditional Korean fermented seafood)-derived Bacillus velezensis Kh2-2 (Kh2-2) postbiotics in vitro, ex vivo, and in vivo. Cell lysates of four Bacillus species were prepared by sonication. In particular, Kh2-2 lysates induced NO production by upregulating iNOS expression in RAW264.7 cells compared with the lysates of B. subtilis Kh2-1, B. vallismortis Kh8-3, and B. amyloliquefaciens Kh3-1. Furthermore, Kh2-2 lysates stimulated immune activation of macrophages by upregulating the NF-κB and MAPK signaling pathways and promoting immune-related cytokine secretion. In the ex vivo study, Kh2-2 lysates stimulated proliferation and polarized Th1 response by inducing the production of IL-2 and IFN-γ and inhibiting IL-10 expression in splenocytes. The in vivo immune-enhancing effects of Kh2-2 lysates and Kh2-2 were further evaluated using a cyclophosphamide (CTX)-induced immunosuppression mouse model. The results showed that oral administration of Kh2-2 lysates improved CTX-induced immunosuppression by enhancing innate and adaptive immunity, stimulating immune-related cytokine secretion, and modulating gut microbiota dysbiosis in mice. Thus, we concluded that Kh2-2 lysates have potential as a functional material for postbiotics with immune-enhancing effects.