More importantly, these biogenic amines are converted into nitrosamine and behave like a carcinogen 12. Particularly, biogenic amines are the most common contamination in soybean paste during fermentation, excessively high levels of which are associated with serious health issues 6. Doenjang is vulnerable to fouling by aflatoxins, biogenic amines and Bacillus cereus which adversely affects human health 6, 11. In our previous study, we successfully made the different microbes-based starter culture, along with the variation in the fermented material composition, to achieve the Doenjang with novel properties that proved efficient to limit the aflatoxin, biogenic amines and pathogenic microbes well below the acceptable level 1, 6, 11. Extensive efforts have been made to improve the Doenjang quality by altering fermentation condition, substrate and microbial inoculum 9, 10. Further, selection of superior quality of basic ingredients (for example, seeds) also have an impact on Doenjang quality which can be achieved by utilizing many modern techniques 8. Hence, the addition of new ingredients has high possibilities to improve the Doenjang quality both in taste and functional property. The nutrient value, taste and texture of Doenjang is highly dependent on fermentation conditions, basic ingredients and involvement of microorganisms 1, 5. Later, the solid portion is separated from the liquid and fermented for several months resulting in Doenjang 6, 7. Subsequently, salt solution is added to Meju and fermented for few more months. Typically, Meju is prepared by soaking, cooking, and crushing soybeans, followed by modelling into solid blocks of defined shape and size where consortia of naturally occurring microbes (bacteria and moulds) grow with the time. Traditionally, Doenjang is prepared using fermented Meju and brine (salt solution). In particular, Doenjang is the most common fermented soy product in Korea with numerous beneficial activities including antioxidant, anti-inflammatory, anticancer and antimutagenic properties 5. Apart from this, it is also consumed in the form of fermented foods such as Kochujang, Chungukjang, Doenjang, soy sauce 1, Sufu 3, Aakhone and Peruyaan 4. Soybeans are the most important legume that contains all the essential amino acids required for human development and can be consumed as such (seeds) or used after processing in the form of soy flour, defatted soy flakes, soy proteins concentrate and soy protein isolates 2. Owing to its high nutritious value, its consumption has increased many folds worldwide in the recent time. Besides proteins, it is a rich source of minerals and vital phytochemicals such as isoflavonoids 1. Soybean ( Glycine max) is a major crop long being used as a chief source of protein in Asian countries. The results collectively suggest that the addition of garlic in Doenjang can improve its nutritional and functional value, resulting in the protection of consumers from protein deficiencies and various stress conditions. In addition, nontoxic amount of anti-nutritional biogenic amines was observed in all the samples. Absence of pathogenic bacteria ( Staphylococcus aureus, Salmonella species and Bacillus cereus) and aflatoxin was observed in Doenjang samples. Furthermore, GGD-10 effectively reduced colony formation and altered mitochondrial membrane potential of HT29 cells. However 10% fresh-garlicsupplemented Doenjang (GGD-10) showed better activity against colorectal adenocarcinoma (HT29) cell line. Among all preparations, 3% freeze-dried garlic-supplemented Doenjang proved the most effective against gastric adenocarcinoma and lung adenocarcinoma with 50% inhibition concentration of 7.66 ± 0.53 mg/mL and 7.82 ± 0.34 mg/mL, respectively. As a measure of nutritive value, a high amount of total free amino acids, 4,290.73 mg/100 g–5,492.94 mg/100 g, was observed in prepared Doenjang. Results showed a considerable antioxidant and anti-proliferative activity of garlic-supplemented Doenjang. Three different forms of garlic, namely, fresh garlic (2%, 6%, 10%), heat-dried (1%, 2%, 3%) and freeze-dried (1%, 2%, 3%), were supplemented in soybean paste to prepare Doenjang and further evaluated for functional, nutritional and safety aspects.
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