Centrifugal Ultrafiltration Coupled With Mass Spectrometry For Screening Enzyme Inhibitors From Traditional Chinese Medicine (TCM)

Tyrosinase(ECl.14.18.1,or TYR)is one of the versatile copper oxidases with oxygenase and oxidase double functions,has been recently attracting significant attentions due to its promising capabilities to synthesis melanin,produce the key enzyme of plaques,and participate in the formation of polyphenol compounds.The overexpression of tyrosine enzyme can lead to the formation of pigmentation,which is the main reason for the browning of fruits and vegetables.Tyrosine inhibitors including kloic acid,hydroquinone and glycoside are capable to inhibit the activity of tyrosinase,effectively prevent and even treat pigmented diseases,thus has been developed for commercial applications.The natural products as a key area for the search of tyrosinase inhibitors are characterized by high efficacy and low toxicity.This article is aim to analysis and identify tyrosinase inhibitors with good curative effect from puerarin and Rhizoma typhonii.Pancreatic lipase(Pancrelipase EC31.1.3,glyceryl ester hydrolysis enzyme)is one of the lipolytic enzymes,which is the key enzyme of the body’s absorption of fat.Pancreatic lipase that is commonly found in plants,animals,and microorganisms,is one of the largest and deepest digestive lipases.Pancreatic lipase participates in the process of fat digestion,absorption and utilization.Therefore,the pancreatic lipase inhibitor is an effective way to reduce the absorption and digestion of exogenous fat in the human body,and show a broad prospect in lipid reduction.Currently,the pancreatic lipase inhibitor orlistat has been successful in clinical treatment of obesity.But this drug can lead to allergies,liver toxicity and kidney toxicity and so on.The natural products as a key area for the search of pancreatic lipase inhibitors are characterized by high efficacy and low toxicity.This article is aim to analysis and identify pancreatic lipase inhibitors with good curative effect from puerarin and lotus leaf.Ultrafiltration centrifugal mass spectrometry(UF-UPLC/Q-TOF/MS)is one of the techniques to screen enzyme inhibitors from the mixture.This method is to use the enzyme based on the principle that small molecules can be combined into complex compounds,using modern chemical analysis method for liquid chromatography-mass spectrometry technology selection of target compounds.Five and seven tyrosine inhibitors were selected by optimizing ultrafiltration centrifugal mass spectrometry.Five and five types of pancreatic lipase inhibitors were selected from the root of the herb and the lotus leaf.Puerarin exhibited much stronger inhibitory activity in vitro than arbutin against tyrosinase.In silico docking study indicates that the interaction between puerarin and tyrosinase mainly derived from the hydrogen bonds.The result provides theoretical basis to develop inhibitors of pancreatic lipase and tyrosinase,which also provides an effective means for the discovery of natural drugs based on target.