A comparative study of compositional, antioxidant capacity, ACE-inhibition activity, RP-HPLC peptide profile and volatile compounds of herbal artisanal cheeses
Introduction
Manufacturing healthy cheese products using herbs is an ever-growing area. The objectives for using herbs are diversifying flavour, extending the shelf stability of the cheese by reducing the microflora reaching a healthful cheese and prevention of physical defects by decreasing the salt concentration (Charles, 2013; Witkowska, Hickey, Alonso-Gomez, & Wilkinson, 2013). The cheese types can vary with the type of milk used, the cheese manufacturing protocol, ripening period and the use of herbs and/or spices, which affect texture and flavour development of cheeses (Fox, Guinee, Cogan, & McSweeney, 2000).
Herbs used in cheese-making are common, especially in Turkey (Hayaloglu & Farkye, 2011; Hayaloglu & Karabulut, 2013), Spain, Portugal (Lomolina, Zannoni, & Di Pierro, 2015) and in some regions of central Italy (e.g., Tuscany, Lazio and Umbria). While a few studies have been provided on proteolytic activities of plant enzymes, not much scientific information is present on the effect of herbs on the bioactivity of peptides during cheese ripening. Some investigations have characterised the use of plant extracts for cheese production with advanced benefits (Fernandez-Salguero, Tejada, & Gomez, 2002; Garcıa et al., 2012). Including herbs and spices and condiments for replacing the salt in cheese to give the perceived saltiness may be successful especially for lightly-flavoured cheeses (Farkye & Guinee, 2017).
Different herbs are used during cheese production from raw or pasteurised milk and the cheese is mostly ripened for 3 months (Yerlikaya & Karagozlu, 2014). There is a rising interest in discovering natural additives that can raise the shelf durability of milk products as well as supporting their flavour and taste characteristics (Tarakci & Deveci, 2019). The herbs most used in dairy products are Mentha sp., Thymus sp., and Allium sp., and these herbs imparted higher bio-preservative and flavour characteristics to the product (Tarakci & Kucukoner, 2008).
ACE-inhibitory (ACE-i) peptides are mainly derived from animal proteins, especially from dairy foods, vegetable proteins and algae (Shu, Shi, Chen, Ji, & Meng, 2017) and used as a possible option to artificial remedies (Wei, Li, & Zhao, 2013). Protein breakdown by proteases was noted to include the production of polypeptides and has been used to improve biological rate or functional quality of cheese proteins and improve flavour property. The ACE-i potency in cheese has been related to the extent of cheese ripening. ACE-i peptides have been isolated from different Finland low-fat cheeses, e.g., Festivo (Ryhänen, Pihlanto-Leppälä, & Pahkala, 2001) and, in Turkey, white-brined cheese (Kocak, Sanli, Anli, & Hayaloglu, 2020; Sahingil, Gokce, Yuceer, & Hayaloglu, 2019).
Climatic-vegetation diversity and geographical location in certain regions of North Macedonia supports production of various traditional cheeses with the use of herbs. The herbs are added to the cheese either during curd mixture or moulding, or the cheese is wrapped with the herbs or applied to the surface of the cheese. There is no previous investigation about the formation of ACE-inhibiting peptides in cheeses produced with herbs and the effect of herbs proteases and peptidases (Sieber et al., 2010).
To date, only a few studies using GC/MS have been conducted for the determination of volatiles generated during herbal cheese ripening (Güler, 2014; Hayaloglu & Karabulut, 2013) and, to the best of our knowledge, no other studies have been performed on the ACE-inhibition activity of the peptides of herbal cheeses. The aim of this work was, therefore, the characterisation of the ACE-inhibition activity of these products. For the present study, we have considered different artisanal cheeses (beaten, Kashkaval and white brined cheeses) combined with different herbs (Mentha longifolia, Origanum vulgare, Thymus longicaulus, Alium sativum, Petroselinum crispum). In this context, the objectives of this study were to determine the gross chemical compositions, peptide profiles, ACE-inhibitory and antioxidant activities and volatile profiles to evaluate the effect of herbs on the ripening of traditional cheeses and to produce a new dairy product with improved nutritional value.
Section snippets
Materials
The herbal cheeses were produced in five different small dairies in Polog Region, North Macedonia. Raw sheep's, cows' or a goats' milk after filtration immediately after milking was coagulated with commercial calf rennet (Maxiren® – chymosin) at milking (36 °C) temperature (approximately 45–60 min). Manufacture of cheeses was conducted at farmhouse cheese-making conditions as given its detail in Fig. 1 and Table 1.
Compositional analysis and assessment of proteolysis
The dry matter, salt, protein, fat and total free amino acid (TFAA) levels in the
Compositional and proteolysis changes
The chemical composition and pH of herbal cheeses were: 46.05–70.40% total solids, 16.16–29.36% protein, 18.25–30.11% fat, pH 4.77–6.21. Use of herbs did not significantly affect (P < 0.05) the dry matter, fat, salt and the soluble nitrogen (except water soluble nitrogen, WSN, in white cheeses) levels of the cheese samples (Table 2). In previous studies on the herbal cheeses, the dry matter, salt and total nitrogen levels were reported as 43.05–58.14; 6.39–7.21 and 2.03–3.99%, respectively. The
Conclusions
Herb incorporation into cheeses did not significantly affect the dry matter, fat, salt and the soluble nitrogen (except for WSN in white and TCA-SN in Kashkaval cheeses) at the end of the ripening period. Cheese with mint incorporated had significantly higher WSN values than the other brined cheese from the same group. Furthermore, thyme and mint incorporated samples had higher WSN values than the control cheese from the same group. Herb antioxidant activity increased total antioxidant levels
Acknowledgements
This study was supported by Erasmus+ Project IFCHEESE, 2017-1-TR01-KA205-044201. The technical assistance of Scientific Research Unit at Inonu University (Malatya, Turkey) is gratefully acknowledged.
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