Application of new Probiotic Bacillus coagulans bc99 in food
Tu Yanhua, Qin Yu, Fang Duan, Zhu Jianguo,
Jiang Tian, Da Xuyang, Zhang Qiuyu
Jiangsu Weikang Biotechnology Co., Ltd. (Suzhou)
Abstract: Bacillus coagulans not only has the function of general probiotics, but also has the characteristics of high resistance to high temperature, acid and bile salt. In this paper, the resistance, function and utility of Bacillus coagulans and its application in different prepackaged foods were discussed. Jiangsu Weikang Biotechnology Co., Ltd. (hereinafter referred to as "Weikang biology") is a leading enterprise providing complete solutions for probiotic products in China. At present, it has realized the industrialized production of Bacillus coagulans bc99, with an annual output of 100 tons of Bacillus coagulans bc99 active bacterial powder, and the number of spores is up to 200-300 billion CFU / g. the probiotic has strong resistance, high temperature resistance and high sporulation rate It can be widely used in all kinds of food.
Key words: Bacillus coagulans bc99, high activity, stress resistance, Bacillus coagulans is a new probiotic that can form spores. It not only has the characteristics very similar to Lactobacillus and Bifidobacterium, but also has the strong stress resistance characteristics because of its spore production characteristics. In 2005, Bacillus coagulans was approved by China's food and Drug Administration (SFDA) to be used as intestinal medicine for human use; in 2012, Bacillus coagulans was approved by the U.S. Food and Drug Administration (FDA) as a safe and edible strain [1-2]; in 2016, Bacillus coagulans was listed in the list of strains available for food by China's health and Planning Commission. At present, Bacillus coagulans has been widely used in food in foreign countries, such as candy, chocolate, beverage, etc.; in China, Bacillus coagulans has also become the target of many food industries, widely used in various fields of food. In recent years, Bacillus coagulans is basically imported, and Bacillus coagulans in domestic market is almost monopolized by foreign countries. After years of careful research, continuous exploration and optimization of technology and introduction of advanced production equipment, Weikang biology has become the first industrial enterprise in China to achieve an annual production capacity of more than 100 tons of Bacillus coagulans bc99 (hereinafter referred to as bc99), which has been put into production, with a spore number of up to 200-300 billion CFU / g and a maximum of up to 500 billion CFU / g. Bc99 has the excellent characteristics of strong resistance to stress, high temperature and pressure, strong resistance to acid and bile salt, high survival rate, easy storage, etc., it can resist the action of gastric acid and digestive enzyme, and can proliferate smoothly in the intestine and play a prebiotic role [3-4]. As a new probiotic, bc99 is becoming a new favorite in the research and development of functional probiotic products. 1. Overview of Bacillus coagulans the earliest record of Bacillus coagulans appeared in the paper published by hammer b.w. in 1915 [5]; in 1932, famous scientist L.M Horowitz wlassowa et al. First used "Lactobacillus sporosum" to describe Bacillus coagulans in their published literature; in 1974, the eighth edition of Berger's Manual of Bacteriology for identification divided it into Bacillus; the bacterium produced L-lactate and coagulant, which was named Bacillus coagulans in 1980.
Figure 1 Bacillus coagulans Coagulansbc99) spore microscopic examination photo: Bacillus coagulans is rod-shaped and blunt at both ends, belonging to Gram-positive bacteria, catalase positive, facultative anaerobic, spore terminal; it can decompose lactose, glucose, sucrose, maltose, mannitol and other sugars to produce L-lactic acid, which is the same type of lactic acid fermentation bacteria; ph2-8 can survive, ph5.5-7.5 is the most suitable growth temperature; it can grow at 30-55 ℃, the most suitable growth temperature Degree 40-50 ℃; biotin and thiamine are the main growth factors to promote its growth [7-8]. Bacillus coagulans can produce antibiotic coagulant, lactic acid, amino acid, short chain fatty acid, vitamin and various digestive enzymes; it is resistant to high temperature, strong acid, high salt and easy to preserve; its physiological effect is similar to that of Lactobacillus and Bifidobacterium [9]. 2. The resistance of Bacillus coagulans. Bacillus coagulans has complex and dense multi-layer spore wall structure and extremely low metabolic activity, which makes it resistant to the adverse effects of the external environment. It has the characteristics of acid and salt resistance, heat and temperature resistance, high sporulation rate and easy storage. When bc99 was treated in 80 ℃ water bath for 5 hours, the cell survival rate was still 50%. When bc99 was treated in 100 ℃ water bath for 2 hours, the cell survival rate was 94%; when bc99 was treated in simulated gastric juice (pH2.5) for 120 minutes, the survival rate was 84.3%; when bc99 was cultured in simulated intestinal juice (0.3%) for 36 hours, the number of viable bacteria was 6.7 × 109 CFU / ml, the sporulation rate reached 89.2% [10]. Spores with excellent stress resistance are the best form of probiotic preparation, which makes bc99 have unique stability advantage in food processing and storage.
Fig. 2 1600 times microscopic examination of bc99 with Gram staining?
Fig. 3 1600 times of bc99 spore staining Photo 3 The important role of Bacillus coagulans in human body the high stress resistance of Bacillus coagulans is the prerequisite for its smooth passage through the gastrointestinal system of human body. Its spores enter the digestive system after eating, and begin to germinate and grow in the gastric juice environment. The spore coat expands, the shape of the spore increases, the water content increases, and the protuberance grows from the spore coat. When entering the duodenum, the spores have germinated and formed a camp The cultured cells then enter the small intestine and begin to multiply in large numbers. They are highly effective in colonizing the intestine and play a prebiotic role [11-12]. Bacillus coagulans can tolerate gastric acid and digestive enzymes to settle in the intestine and restore physiological metabolism, and play a prebiotic role by secreting a variety of metabolites and cooperating with other probiotics in the intestine.
Three point one It can regulate the microecological balance of intestinal flora, inhibit the growth of pathogenic bacteria, coagulate bacillus is a facultative anaerobic bacteria, which consumes free oxygen after settling in the intestinal tract and creates an anaerobic intestinal environment; it secretes antibacterial substances such as lactic acid and coagulant, inhibits the growth of aerobic harmful bacteria, promotes the proliferation of anaerobic probiotics such as lactic acid bacteria and bifidobacteria, recovers and maintains the intestinal tract Microecological balance of flora. Lu Fuping [13] found that Bacillus coagulans has a certain inhibitory effect on common intestinal pathogens such as E Marrec et al. [14] found that Bacillus coagulans can secrete antimicrobial peptide - coagulant. Hyronimus et al. [15] found that coagulant has strong stability and has inhibition effect on Enterococcus, leucococcus, Listeria and Micrococcus. It can be widely used in food industry to play the role of intestinal regulation. In addition, Bacillus coagulans can also produce diacetyl, organic acids and other metabolites, which can reduce the production of harmful substances such as amines, improve the micro ecological environment in the intestinal tract, and reduce the occurrence of intestinal diseases [16-17]. 3.2 it can promote intestinal digestion and absorption, improve nutrition utilization rate. Bacillus coagulans has a very rich enzyme system, which can secrete a variety of digestive enzymes, such as β - galactosidase, amylase, lipase, protease, etc., degrade macromolecular substances into micromolecular substances, promote the digestion and absorption of nutrients, and improve the nutrition conversion and utilization rate of food [18]. Batra et al. [19] found that Bacillus coagulans can secrete β - galactosidase, decompose lactose in milk into glucose and galactose, effectively alleviate lactose intolerance, and improve the digestion and absorption of milk. Keller D et al. [20] found that the plant protease secreted by Bacillus coagulans can degrade the plant protein into small peptide and amino acid, and promote intestinal absorption and metabolism. In addition, Bacillus coagulans produces vitamins, short chain fatty acids and other metabolites in the process of growth, reproduction and colonization, which can stimulate intestinal peristalsis, promote the digestion and absorption of nutrients, reduce the accumulation of toxins, and improve the nature of feces. Ara et al. [21] found that Bacillus coagulans can reduce intestinal ammonia value, improve defecation frequency, improve fecal odor and color, and improve intestinal metabolic environment. 3.3 activation of immune cells, enhancement of immunity and slowing down of Bacillus coagulans can normalize the quantitative parameters of immune system (the number of spleen lymphocytes, macrophages and T-lymphocytes) and the functional activity of cells, and improve the function of host immune system. By regulating cytokines to inhibit reactive oxygen species, enhance phagocytic activity, promote the normal immune response of digestive tract mucosa, so as to improve the function of host immune system. Kodali et al. [17] found that Bacillus coagulans can inhibit the formation of ROS induced by polymorphonuclear leukocytes (PMN, mainly neutrophils), increase the phagocytic activity of neutrophils, and improve the immunity of the body by inhibiting ROS and enhancing phagocytosis. In addition, Bacillus coagulans can also induce the expression of CD69 in NK cells, activate the marker cd3-cd56 + T cells, natural killer (NK) cells and other cells [22], and promote the immune regulation of the body. Bacillus coagulans can activate the host immune cells, enhance the host immune system through the medium of cell source, increase anti-inflammatory cytokines, down regulate inflammatory immune factors such as interferon (IFN - γ), interleukin (IL), etc., and relieve inflammation without changing the normal regulatory cytokines. Sudha et al. [23] have found that Bacillus coagulans can inhibit the expression of COX-2, IL-12, TNF - α and IFN - γ in inflammatory cells, and thus play a role in alleviating inflammation. With the development of social economy and the improvement of people's living standard, how to live a healthy life has gradually become a hot topic in daily life. Probiotics, as a functional food, are more and more popular. The new probiotics Bacillus coagulans showed strong resistance to heat, temperature and stability, which greatly enriched its application in the food industry.
4.1 with the upgrading of residents' consumption, the leisure food market in China is gradually upgrading to a younger, higher-end and healthier market. Under the trend of "the fourth meal", the sales volume of leisure snacks has exceeded trillion, and will maintain a compound growth rate of more than 20%. From the change trend of market scale in recent years, China's leisure food industry will show a long-term stable growth trend. In addition to the function of common probiotics, Bacillus coagulans also has the characteristics of high temperature resistance and storage resistance.
4.1.1 application in candy Wang Zhen et al. [24] applied Bacillus coagulans to candy, adding cocoa powder at 110 ℃ and whole milk powder at 100 ℃ had no effect on the survival of Bacillus coagulans. The probiotics developed by Weikang biology coagulate the soft sugar of Bacillus, each bag is specially added with 12 billion active bc99, which can be stored at room temperature, and can maintain excellent stability within the shelf life. Adding Bacillus coagulans to chocolate with mellow and silky taste can maintain its high activity, promote body digestion and maintain intestinal microecology balance.
4.1.2 application in nuts as a healthy and green snack, nuts are widely favored by consumers. Driven by both supply and demand, domestic nuts grow rapidly. According to Sullivan's statistics, in 2018, the proportion of stir fried nuts in leisure food is about 10%, and the nut industry is also undergoing three major trends: from nucleated to non nucleated, from single product to composite, from rough processing to deep processing. The probiotic nut, with the characteristics of natural, leisure, nutrition, health and delicious, is advancing towards high quality, high beauty and high cost performance, precisely meeting the health needs of the market