*Article* **Study of the Lipolysis Effect of Nanoliposome-Encapsulated** *Ganoderma lucidum* **Protein Hydrolysates on Adipocyte Cells Using Proteomics Approach**

**Sucheewin Krobthong 1, Yodying Yingchutrakul 2, Wonnop Visessanguan 3, Thanisorn Mahatnirunkul 4, Pawitrabhorn Samutrtai 5, Chartchai Chaichana 6, Phakorn Papan 7 and Kiattawee Choowongkomon 1,8,9,\***


**Abstract:** 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 nanoliposomal 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

**Keywords:** RSM; Lingzhi; hypolipidemic activity; peptides; 3T3-L1
