Ganoderma lucidum is an edible fungus which typically used in functional food products because of the potential enhancement of health. Many hydrolysates from G. lucidum exhibit various biological functions. Using LC-MS/MS, the peptide sequence was predicted based on the de novo sequencing algorithm and P1-peptide (VDLPTCKGF) was identified. The peptide was modified by shortening C-terminal residue one at a time until three residues remained, and chemically synthesized all modified peptides by SPPS. Seven-synthetic peptides were purified and molecularly evaluated by HPLC and LC-MS. The antioxidant capacity of these peptides was evaluated, while also the intracellular-ROS was detected to validate the antioxidant activity. This study showed that 3 of 7 synthetic peptides had in vitro antioxidant activity and suppressed intracellular-ROS. Among all modified peptides, VDLPTC exhibited the highest antioxidant capacity and revealed intracellular-ROS suppression. This evidence reflects their radical-scavenging activities and provides the rationale for the development of novel functional food ingredients.

Grammatophyllum speciosum is the largest orchid species and a well-known traditional medicinal plant. Due to skin aging, natural products that inhibit this process can attract the attention of consumers and scientists because radical-scavenging activity, collagenase inhibition, and inflammatory suppression are valuable in dermatological applications. This study investigated the phytochemicals in G. speciosum leaves extracts that have cosmeceutical potentials, including radical-scavenging, anticollagenase, and anti-inflammatory abilities. G. speciosum leaves were extracted using water-based extraction methods. High-resolution mass spectrometry was used to identify the phytochemicals in the extracts. Fibroblast and keratinocyte cell cytotoxicity was determined. Antioxidant abilities were measured using DPPH and ABTS assays. The effect of the extracts on nitric oxide (NO) in macrophage cells was investigated. ELISA of the collagenase enzyme was determined. A total of 721 annotated metabolites were identified in the extracts. Vitexin and orientin were the most abundant metabolites. Cell viability was >80% in both cell lines when the extract concentration was <1 mg/mL. The IC₅₀ values for DPPH and ABTS were 56 and 117 μg/mL, respectively. Furthermore, the extracts revealed that NO and collagenase activity were suppressed by 42% and 23%, respectively. The extracts can suppress ROS, inflammatory, and collagenase activities without causing fibroblast and keratinocyte cell death. Thus, this study provides information on metabolites in G. speciosum leaves, which is promising as cosmeceuticals or pharmaceuticals with anti-inflammatory and anti-collagenase activities.

Excessive lipid accumulation is a serious condition. Therefore, we aimed at developing safe strategies using natural hypolipidemic products. Lingzhi is an edible fungus and potential lipid suppression stimulant. To use Lingzhi as a functional hyperlipidemic ingredient, response surface methodology (RSM) was conducted to optimize the time (X₁) and enzyme usage (X₂) for the hydrolysate preparation with the highest degree of hydrolysis (DH) and % yield. We encapsulated the hydrolysates using nanoscale liposomes and used proteomics to study how these nano-liposomal hydrolysates could affect lipid accumulation in adipocyte cells. RSM analysis revealed X₁ at 8.63 h and X₂ at 0.93% provided the highest values of DH and % yields were 33.99% and 5.70%. The hydrolysates were loaded into liposome particles that were monodispersed. The loaded nano-liposomal particles did not significantly affect cell survival rates. The triglyceride (TG) breakdown in adipocytes showed a higher TG increase compared to the control. Lipid staining level upon the liposome treatment was lower than that of the control. Proteomics revealed 3425 proteins affected by the liposome treatment, the main proteins being TSSK5, SMU1, GRM7, and KLC4, associated with various biological functions besides lipolysis. The nano-liposomal Linzghi hydrolysate might serve as novel functional ingredients in the treatment and prevention of obesity

Leishmaniasis is a tropical disease caused by Leishmania parasites, which are transmitted through the bites of infected sandflies. We focused on the emergence of leishmaniasis in Thailand caused by a species (Leishmania orientalis). Treatment by chemotherapy is not effective against L. orientalis. Hence, we intended to solve this issue using a proteomics approach to investigate protein profiles and in silico analysis for the identification of antigenic proteins from L. orientalis, Leishmania martiniquensis, and Leishmania donovani. Using principal component analysis (PCA), protein profile comparisons indicated that different species of Leishmania are different at the protein level. Proteomics analysis identified 6099 proteins. Among these proteins, 1065 proteins were used for further analysis. There were 16 proteins that were promising candidates for therapeutic aspects as they were abundantly expressed and common to all species. In silico analysis of protein’s antigenicity revealed that eight proteins had the potential for the development of antigenic molecules. Protein profile information and these antigenic proteins may play key roles in the pathogeny of leishmaniasis and can be used as novel therapeutic targets against leishmaniasis in the future.

Methicillin-resistant Staphylococcus aureus (MRSA) is listed as a high-priority pathogen because its infection is associated with a high mortality rate. It is urgent to search for new agents to treat such an infection. Our previous study isolated a soil bacterium (Brevibacillus sp. SPR-20), showing the highest antimicrobial activity against S. aureus TISTR 517 and MRSA strains. The present study aimed to purify and characterize anti-MRSA substances produced by SPR-20. The result showed that five active substances (P1–P5) were found, and they were identified by LC-MS/MS that provided the peptide sequences of 14–15 residues. Circular dichroism showed that all peptides contained β-strand and disordered conformations as the major secondary structures. Only P1–P4 adopted more α-helix conformations when incubated with 50 mM SDS. These anti-MRSA peptides could inhibit S. aureus and MRSA in concentrations of 2–32 μg/mL. P1 (NH₂-VVVNVLVKVLPPPVV-COOH) had the highest activity and was identified as a novel antimicrobial peptide (AMP). The stability study revealed that P1 was stable in response to temperature, proteolytic enzymes, surfactant, and pH. The electron micrograph showed that P1 induced bacterial membrane damage when treated at 1× MIC in the first hour of incubation. The killing kinetics of P1 was dependent on concentration and time. Mechanisms of P1 on tested pathogens involved membrane permeability, leakage of genetic material, and cell lysis. The P1 peptide at a concentration up to 32 μg/mL showed hemolysis of less than 10%, supporting its safety for human erythrocytes. This study provides promising anti-MRSA peptides that might be developed for effective antibiotics in the post-antibiotic era.

Ganoderma lucidum or Lingzhi is a fungus species widely known as a traditional medicine. Exploring the beneficial peptides by hydrolysis using pepsin and trypsin has been extensively performed to identify new bioactive natural products. A multifunctional peptide that expresses potential scavenging activity and tyrosinase inhibition is valuable in therapeutic and cosmetic applications. This study aimed to identify and investigate the effects of a novel multifunctional peptide from Lingzhi on the melanogenic enzymes in melanoma cells by a targeted-proteomics approach. The multifunctional peptide was de novo sequenced by LC-MS/MS to be NH₂-PVRSSNCA-CO₂H (octapeptide). This sequence was chemically synthesized by solid-phase peptide synthesis (SPPS). The antioxidant ability of the synthesized octapeptide was measured by the DPPH, ABTS, and FRAP assays. The results showed that the peptide exhibited an antioxidant activity equal to 0.121 ± 0.01 mg equivalent to ascorbic acid, 0.173 ± 0.03 mg equivalent to gallic acid, and 2.21 ± 0.23 mM equivalent to FeSO₄, respectively, which is comparable to these well-known antioxidants. The proteomics approach identified a total of 5804 proteins and several pathways involved in the effects of the octapeptide in melanoma cells. Targeted proteomics revealed three specific proteins associated with pigmentation including Rab29, Dct, and Tyrp1. The Rab29 and Dct were upregulated whereas Tyrp1 was downregulated in the octapeptide treatment group. These findings could be used in the understanding of the molecular functions of the multifunctional octapeptide on melanogenic enzymes, supporting its potential as a therapeutic and cosmetic ingredient.