45 Research Article Fatty Acid Composition, Antinutritional Factors, and Oligosaccharides Concentration of Hawaijar (An Ethnic Fermented Soyfood of India) As Afected by Genotype and Bacillus Subtilis Strain L Sophia Devi 2 , Vineet Kumar 1* , Anita Rani 1 , Trupti Tayalkar 1 , Priyanka Mittal 1 , Aseem Kumar Anshu 1 , Thangjam Anand Singh 2 1 ICAR-Indian Institute of Soybean Research, Indore, M.P., India 2 Central Agricultural University, Imphal, Manipur, India *Corresponding Author: Vineet.Kumar@icar.gov.in Abstract: Hawaijar is an ethnic fermented soy product from the North-East region of India. The information on the effect of soybean genotype and the Bacillus subtilis strain used for inoculation for preparing the product on the fatty acid composition, the level of antinutritional factors, such as Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI), and oligosaccharides is not available. In the present study, Hawaijar was prepared from two soybean genotypes using two different strains of Bacillus subtilis and investigated for the concentration of the above-mentioned biomolecules. The results showed the signifcant effect of genotype used on oleic, linoleic, and α-linolenic acid of the product. KTI and BBI were found absent in Hawaijar prepared from these two genotypes and two strains. Flatulence-inducing factors like raffnose and stachyose were found in very low concentration irrespective of genotype and strain. Retention of sweetness-imparting sucrose and maltose concentration in Hawaijar was genotype-dependent. Keywords: Hawaijar, Fermented soybean, Fatty acid composition, Protease inhibitors INTRODUCTION Globally, the inclusion of soybean in daily diet is being recommended to meet the daily requirement of protein and avail numerous other health benefts derived from nutraceutical molecules reported to be present in soybean (Kumar et al., 2010a; Hendrich et al., 2017). Apart from blending soybean four with wheat four to prepare Indian fat bread (chapati), which is considered the easiest mode of incorporation of soybean in a daily diet, several unfermented soy products, namely, soy chunks, soy granules, tofu and soy milk are available in the market across the country. However, in the North-Eastern region of the country which includes eight states, namely, Nagaland, Mizoram, Manipur, Arunachal Pradesh, Meghalaya, Assam, Tripura and Sikkim, several naturally fermented soy products like Aakhone/axone (Nagaland), Bekanthu (Mizoram), Hawaijar (Manipur), Preuyaan (Arunachal Pradesh), Turangbai (Meghalaya) and Kinema (Sikkim) are in the diet of ethnic population of the region since several centuries (Tamang, 2015; Singh et al., 2019). Hawaijar is an indigenous traditional fermented soybean product of Manipur state and also a staple in every household. It is alkaline and sticky in appearance. It can be consumed directly or used as a favoring agent in vegetables throughout the year and its addition make the vegetables soft and tasty. Traditionally, the preparation of fermented soybean, Hawaijar, involves overnight soaking of soybean followed by washing under running tap water. Then, they are boiled well in a pressure cooker till the seeds become soft and washed with hot water. After draining out excess water, the boiled seeds are wrapped in healthful clean Ficus hispida leaves and shifted in a bamboo basket covered with lid, called ‘lubak’, which is covered with clean banana leaves. Subsequently, the basket is kept under the sun for fermentation. This is to maintain the high temperature required for Hawaijar formation. The palatable stage of fermented soybean reaches in 3-5 days. After fermentation, the soybean seeds develop a brown colour, sticky texture and a distinctive fermented odour and taste. Unlike other fermented soy-products from East and South-East Asian countries which have been evaluated for nutritional properties (Dajanta 2011; Jing et al., 2017). Hawaijar has not yet been biochemically characterized. In the present investigation, Hawaijjar was prepared in the laboratory from two soybean genotypes, namely, a commercial cultivar JS335 and a landrace being grown in the North-Eastern region of India, using two different strains of Bacillus subtilis. Hawaijar products prepared were subsequently analyzed for the fatty acid composition, concentration of antinutritional factors, namely, Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI), and oligosaccharides such as, sucrose and maltose which impart sweetness, and raffnose and stachyose which are responsible for fatulence associated with soy products. MATERIALS AND METHODS Genotypes used in this study were commercial cultivar JS335 and landrace of soybean (small seeded). Strain S and H of Bacillus subtilis were used in the preparation of Hawaijar. Chemicals: Petroleum ether, sodium methoxide, acrylamide, bis-acrylamide, ammonium persulphate, tetramethylethylenediamine (TEMED), bromophenol blue dye, Kunitz trypsin inhibitor standard protein, coomassie brilliant blue dye, methanol, water, acetic acid, tris buffer, phosphate buffered saline, Bowman-Birk inhibitor standard protein, tween 20, bovine serum albumin, Bowman-Birk primary antibody, anti-mouse antibodies, p-nitrophenyl phosphate (pNPP), sodium hydroxide (NaOH), acetonitrile, https://journal.ugm.ac.id/ifnp IFNP, Vol. 17, No. 2 (2020) ISSN 2597-9388 DOI: 10.22146/ifnp.52241