Contents lists available at ScienceDirect Food Chemistry journal homepage: www.elsevier.com/locate/foodchem Use of asparaginase for acrylamide mitigation in cofee and its infuence on the content of cafeine, chlorogenic acid, and cafeic acid Carla Levi Oliveira Corrêa a , Edmar das Merces Penha b , Marianna Ramos dos Anjos b , Sidney Pacheco b , Otniel Freitas-Silva b , Aderval Severino Luna a , Leda Maria Fortes Gottschalk b, ⁎ a Chemical Engineering Department, Rio de Janeiro State University, Rio de Janeiro, Brazil b Embrapa Food Agroindustry, Rio de Janeiro, Brazil ARTICLEINFO Keywords: Cofee Asparaginase Acrylamide Cafeine Chlorogenic acid Cafeic acid ABSTRACT A factorial design with a duplicate in the central point was used to investigate the efect of treating arabica cofee beans with asparaginase. The investigated factors were enzymatic load (1000 and 5000 ASNU/Kg), water percentage (30 and 90%), and hydrolysis time (1 and 3 h). The acrylamide content was determined by UPLC- MS/MS, and the cafeic acid, chlorogenic acid and cafeine concentrations were determined by HPLC-DAD. The statistical analysis was carried out in the R platform using RStudio graphical interface. The results indicated the importance of cofee bean pretreatment with steam, and that the enzyme load reduced the acrylamide content to 65 mg/kg in cofee beans. The predicted reduction was obtained with hydrolysis time of 2 h, water content of 90%, and asparaginase load of 5000 ASNU/kg. The asparaginase treatment did not infuence the major bioactive compounds in cofee. 1. Introduction The use of enzymes in cofee processing has been increasing to improve the fnal product, such as asparaginase to decrease the content of acrylamide (Porto et al., 2019; Xu et al., 2016). Two decades ago, a group of researchers from Sweden observed the presence of acrylamide in carbohydrate-rich foods when roasted or fried, with cereals, pota- toes, and cofee possibly being signifcant sources of ingestion (Tareke et al., 2002; Svensson et al., 2003). At food processing temperatures above 120 °C, asparagine can react with reducing sugars via the Maillard reaction to produce acrylamide (Stadler et al., 2002; Tareke et al., 2002; Rannou et al., 2016; Zilic et al., 2020). Acrylamide has several toxic properties, including genotoxicity, carcinogenicity and neurotoxicity. The International Agency for Re- search on Cancer (IARC, 1994) classifed acrylamide as a probable human carcinogen (Group 2A), and the Scientifc Committee on Food (SCF, 2002) reported it is genotoxic. Research shows that the main foods that contribute to consumer exposure to acrylamide are potato chips (6–46%), cofee (13–39%), bakery products and sweet cookies (10–20%), and bread and toast (10–30%) depending on the type of processing used (Dias et al., 2009). Cofee can contribute signifcantly to the total acrylamide content of the diet (Porto et al., 2015). The intake of acrylamide through cofee varies widely geographically. For example, cofee consumption re- presents about 30% and 40% of total acrylamide in the adult diets in Norway and Sweden, respectively, and 8% and 13% in the Netherlands and United States (Friedman & Levin, 2008). However, the formation of acrylamide in food can be minimized by the use of two enzymes: as- paraginase, which catalyzes the hydrolysis of asparagine, one of its main precursors, into aspartic acid and ammonia by the hydrolysis of the asparagine side-chain amide group (Arima et al., 1972; Hendriksen, 2009); and acrylamidase, a type of amidase that can turn acrylamide into acrylic acid (Cha & Chambliss, 2011). Regarding the use of asparaginase (EC 3.5.1.1), there are two commercially available products for acrylamide reduction in the food industry. These commercial enzymes are produced by Aspergillus spp. and are widely used in commercial products, with safety indicated by the Joint FAO/WHO Expert Committee on Food Additives (JECFA, 2007; Xu et al, 2016). Furthermore, these enzymes are deactivated during the heating process, ensuring their safe application in the food industry (Hendriksen et al., 2009). Asparaginase has received “gen- erally recognized as safe” (GRAS) status from the US Food and Drug Administration. It is currently used in several countries, including the United States, Australia, New Zealand, China, Russia, Mexico, and https://doi.org/10.1016/j.foodchem.2020.128045 Received 13 July 2020; Received in revised form 3 September 2020; Accepted 4 September 2020 ⁎ Corresponding author. E-mail addresses: edmar.penha@embrapa.br (E. das Merces Penha), marianna.anjos@embrapa.br (M.R. dos Anjos), sidney.pacheco@embrapa.br (S. Pacheco), otniel.freitas@embrapa.br (O. Freitas-Silva), leda.fortes@embrapa.br (L.M.F. Gottschalk). Food Chemistry 338 (2021) 128045 Available online 09 September 2020 0308-8146/ © 2020 Elsevier Ltd. All rights reserved. T