Application of Milk-Clotting Protease from Aspergillus oryzae DRDFS13 MN726447 and Bacillus subtilis SMDFS 2B MN715837 for Danbo Cheese Production

Abstract

This study aimed to investigate the efficiency, biochemical composition, and sensory quality of Danbo cheese produced using proteases derived from the fungus and bacterium compared to the commercial product. A fungal enzyme from Aspergillus oryzae DRDFS13MN726447 and a bacterial enzyme from Bacillus subtilis SMDFS 2B MN715837 were produced by solid-state and submerged fermentation, respectively. The crude enzyme from A. oryzae DRDFS13 and B. subtilis SMDFS 2B was partially purified by dialysis and used for Danbo cheese production using commercial rennet (CHY-MAX® Powder Extract NB, Christian Hansen, 2235 IMCU/g) as a control. The Danbo cheese produced using dialyzed fungal enzyme (E1) (267 U/mL), dialyzed bacterial enzyme (E2) (522 U/mL), and commercial rennet (C) were analyzed for body property, organoleptic characteristics, and proximate and mineral composition when fresh and after 2 months of ripening. There was no significant difference in the cheese yield (C = 9 kg, E1 = 8.6 kg, and E2 = 8.9 kg) among the three treatments. The body properties of Danbo cheese produced with the fungal enzyme (E1) were firm and acceptable as the control (C), whereas the Danbo cheese produced by bacterial enzymes has shown a watery body. The overall organoleptic characteristics of Danbo cheese produced by the fungal enzyme (5.3) were similar to control cheese produced by commercial rennet (5.5). Both cheese types were significantly different in organoleptic properties from Danbo cheese produced by the bacterial enzyme (4.9). There was no significant difference (p>0.05) in the proximate composition between the ripened Danbo cheese produced by fungal enzyme and the control cheese except for crude protein content. However, the ripened cheese products showed a significant difference in their mineral composition except for sodium. In conclusion, this study demonstrated that the fungal enzyme from Aspergillus oryzae DRDFS 13 is more appropriate for Danbo cheese production than the bacterial enzyme from Bacillus subtilis SMDFS 2B. However, it requires further application of the enzymes for the production of other cheese varieties.

The catalytic and kinetic characterization of Bacillus subtilis MK775302 milk clotting enzyme: comparison with calf rennet as a coagulant in white soft cheese manufacture

“…It is well known that the higher acidity in the milk decreases the moisture content [ 49 , 50 ]. Other authors also found that the moisture content was decreased in ripened Danbo cheese [ 51 ].…”

“…The white soft cheese made from the partially purified MCE had higher TA than control cheese, and this might be due to the accumulation of lactose degradation products such as lactic acid and volatile fatty acids [ 7 , 48 , 51 ]. During the storage period, TA increased significantly ( P ≤ 0.05), and this might be due to lactose fermentation in the presence of starter cultures which results in the production of some acidic compounds and might be also due to the degradation of protein and fat [ 11 , 48 , 50 – 52 ].…”

Review: Harvesting therapeutically important bioactive compounds from possible poisonous Clitocybe species

“…The compounds present in Clitocybe have been confirmed to have a wide range of beneficial effects, including antitumor, bacteriostat, antifungal, antiviral, antiinflammatory, antioxidant, antiaging, antidiabetic, hypoglecemic, immune stimulant, anti-proliferative, neuroprotective, cardio-protective and chemopreventive properties (Zheng et al, 2010;Chen et al, 2013). It has also found its way into the food industry with notable characteristics such as milk-coagulant, flavorings agent, organic food preservative and yeast culture (Mamo et al, 2020). Other known applications, such as biological pest control, have also been confirmed (Kamikawa et al, 1988;Rahi and 2016).…”

“…Aspartic proteases (APs) (EC 3.4.23) proteolytic enzymes have been isolated from Clitocybe nebularis (Sabotič et al, 2009). Aspartic proteases (APs) have been used in large amounts in the dairy industry as milkcoagulating enzymes, in the manufacture of cheese and in the food industry (Mamo et al, 2020). It has been reported that the worldwide turnover of industrial enzymes isolated from mushrooms is $1 billion.…”

Plant-associated endophytic fungi as potential bio-factories for extracellular enzymes: Progress, Challenges and Strain improvement with precision approaches

“…Additionally, the interfering pigments from the skin get easily removed by the action of proteases to produce clean hide (Zambare et al, 2011). Acidic proteases have tremendous application in the dairy industry serving for making different varieties and textures of cheese (Mamo et al, 2020). Recently, reports are now made available about the necessity of proteinases for nutritional value in dairy products (Mir Khan & Selamoglu, 2020).…”