The physico-chemical alteration of lovastatin and enhanced antioxidant effect of Bacillus subtilis fermented-red yeast rice product Sang Il Gum a , Phuc Anh Nguyen a , Jong Rok Lee b , Yeong Hwan Han c , Min Kyung Cho a,⇑ a Department of Pharmacology, College of Oriental Medicine, Dongguk University, Kyungju 780-714, South Korea b Department of Pharmaceutical Engineering, Daegu Haany University, Daegu 712-715, South Korea c Department of Medical Bioscience, College of Natural Science, Dongguk University, Kyungju 780-714, South Korea article info Article history: Received 5 October 2016 Received in revised form 10 March 2017 Accepted 4 April 2017 Available online 6 April 2017 Keywords: Bacillus subtilis Red yeast rice Fermentation 3(2H)-benzofuranone Antioxidant activity abstract Red yeast rice product (RYP) has been used as a food supplement because of its lipid lowering, and in food additives as a natural colorant. Lovastatin of RYP is a hypolipidemic commercial drug. To enhance the beneficial effects of RYP, we performed a bioconversion with Bacillus subtilis. This B. subtilis-fermentation process of RYP increased the ratio of the active open-hydroxyl acid form and the prodrug lactone form of lovastatin, which is a potent cholesterol synthesis inhibitor. 3(2H)-benzofuranone was newly produced in the fermented red yeast rice product (FRYP) as analyzed by GC–MS. FRYP increased the free radical scav- enging activity compared with RYP. FRYP blocked xanthine oxidase (XO)-induced oxidative cytotoxicity and inhibited the H 2 O 2 -induced intracellular ROS in cells. This is the first study to illustrate that B. subtilis-fermented FRYP is useful for facilitating the alteration in the physico-chemical property of lovastatin and enhancing antioxidant activity, which may have greater pharmacological activity. Ó 2017 Elsevier Ltd. All rights reserved. 1. Introduction Red yeast rice fermented by Monascus species is believed to ameliorate indigestion and hyperlipidemia as a traditional therapy in Asia (Ma et al., 2000; Xiong, Wang, Li, Zhang, & Li, 2015). This microbial product is widely used for the coloring of foodstuffs (Ma et al., 2000). Most researches on red yeast rice has focused on the production of monacolin K, which is a cholesterol synthesis inhibitor leading to the upregulation of low-density lipoprotein (LDL) receptors in the liver and thus decreasing LDL content in the blood (Pan et al., 1990). Monacolin K produced by fermentation was the first pharmaceutical product in the market and then syn- thetic statin drugs has developed. Monacolin K, also known as lovastatin, is administered as an inactive lactone closed ring form, which must be metabolized to a b-hydroxyl acid active open ring form that acts as a reversible competitive inhibitor of HMG-CoA reductase in the liver (Hamelin & Turgeon, 1998; Pan et al., 1990). Lovastatin and other statins have been further studied for their pleiotropic effects independent of their HMG-CoA reductase (Michalik et al., 2013; Mira et al., 2013). The cellular dysfunction caused by reactive oxygen species (ROS) plays a pivotal role in the pathogenesis of liver injury, cardio- vascular disease and cancer (Cesaratto, Vascotto, Calligaris, & Tell, 2004; Soory, 2009). A variety of antioxidants against oxidative damage have been studied to maintain cellular redox-status home- ostasis (Cesaratto et al., 2004; Trueba, Sanchez, & Giuliani, 2004). Diverse antioxidants protect against acetaminophen-induced hep- atotoxicity, which is caused by the reactive metabolic intermediate NAPQI (Gum & Cho, 2013; Oz et al., 2004). The intake of antioxi- dants such as polyphenol and flavonoid compounds found in food products has been suggested as a strategy to reduce oxidative damage (Soory, 2009). Fermented products have been reported to significantly increase the contents of bioactive phenolic com- pounds with a decrease in the amount of glycoside forms in natural foods (Martins et al., 2011). Fermentation using microorganisms is known as a core tech- nique to produce enhanced value in the field of the food, pharma- ceutical and cosmetic industries. Bacillus subtilis has a GRAS (generally regarded as safe) status and produces a variety of pro- teases, b-glucosidase and other enzymes, which are able to degrade a variety of natural substrates producing secondary metabolites dependent on these substrates (Westers, Westers, & Quax, 2004). B. subtilis is the main species of fermentation to produce Natto, Korean Chungkookjang, Indian Kinema and fermented soybean products (Ashiuchi et al., 2001; Yin, Lin, & Jiang, 2010). These http://dx.doi.org/10.1016/j.foodchem.2017.04.023 0308-8146/Ó 2017 Elsevier Ltd. All rights reserved. Abbreviations: FRYP, fermented red yeast rice product; ROS, reactive oxygen species; RYP, red yeast rice product; XO, xanthine oxidase. ⇑ Corresponding author. E-mail address: mkcho@dongguk.ac.kr (M.K. Cho). Food Chemistry 232 (2017) 203–209 Contents lists available at ScienceDirect Food Chemistry journal homepage: www.elsevier.com/locate/foodchem