Amino acid profiles of lactic acid bacteria, isolated from kefir grains and kefir starter made from them

Abstract

The characteristics of cell growth, lactic acid production, amino acid release and consumption by single-strain cultures of lactic acid bacteria (isolated from kefir grains), and by a multiple-strain kefir starter prepared from them, were studied. The change in the levels of free amino acids was followed throughout the kefir process: single-strain kefir bacteria and the kefir starter ( Lactococcus lactis C15–1% + Lactobacillus helveticus MP12–3% + ( Streptococcus thermophilus T15 + Lactobacillus bulgaricus HP1 = 1:1)–3%) were cultivated in pasteurized (92 °C for 20 min) cow's milk (3% fat content) at 28 °C for 5 h (the kefir starter reached pH 4.7) and subsequently grown at 20 °C for 16 h; storage was at 4 °C for 168 h. The strain L. helveticus MP12 was unrivaled with respect to free amino acid production (53.38 mg (100 g) − 1 ) and cell growth (17.8 × 10 8 CFU ml − 1 ); however, it manifested the lowest acidification activity. L. bulgaricus HP1 released approximately 3.7 times less amino acids, nearly 5 times lower cell growth, and produced about 1.2 times more lactic acid. S. thermophilus T15 demonstrated dramatically complex amino acid necessities for growth and metabolism. With L. lactis C15, the highest levels of growth and lactic acid synthesis were recorded (18.3 × 10 8 CFU ml − 1 and 7.8 g l − 1 lactic acid at the 21st hour), and as for free amino acid production, it approximated L. bulgaricus HP1 (17.03 mg (100 g) − 1 maximum concentration). In the L. lactis C15 culture, the amino acids were used more actively throughout the first exponential growth phase (by the 10th hour) than during the second growth phase. The unique properties of the L. helveticus MP12 strain to produce amino acids were employed to create a symbiotic bioconsortium kefir culture, which, under conditions of kefir formation, enhanced lactic acid production and shortened the time required to reach pH 4.7; intensified cell growth activity, resulting in a respective 90- and 60-fold increase in the concentration of lactobacilli and cocci in the mixed culture compared to individual cultures; and accumulated free amino acids in the final kefir with higher total concentrations (56.88 mg (100 g) − 1 ) and an individual concentration of essential amino acids (1.5 times) greater than that of yogurt.