STLO, INRAE, Institut Agro, 35042, Rennes, France.
Appl Microbiol Biotechnol. 2021 May;105(9):3485-3494. doi: 10.1007/s00253-021-11273-3. Epub 2021 Apr 22.
Propionibacterium freudenreichii is a beneficial food-grade actinobacterium, widely implemented, and thus consumed, in various food products. As the main application, P. freudenreichii is used as a cheese-ripening starter, mostly in hard type cheeses. Indeed, during manufacture of "Swiss-type" cheeses (or opened-body cheeses), the technological process favors propionibacteria growth, as well as the corresponding propionic fermentation. This leads to the characteristic flavor of these cheeses, through the release of short chain fatty acids and through lipolysis, as well as to their specific texture. To fulfil this ripening, massive amounts of propionibacteria are industrially produced, dried and stored, prior to cheese making. Furthermore, P. freudenreichii is commercialized in various probiotic food supplements aiming at preserving intestinal health and comfort, in line with its ability to produce beneficial metabolites (short chain fatty acids, vitamins), as well as immunomodulatory compounds. Other industrial applications of P. freudenreichii include the production of food-grade vitamins of the B group, of trehalose, of conjugated linoleic acid, and of biopreservatives. For these different applications, maintaining survival and activity of propionibacteria during production, drying, storage and finally implementation, is crucial. More widely, maintaining live and active probiotic bacteria represents a challenge as the market for probiotic products increases. Probiotic bacteria are, for a bulk majority, freeze-dried, but spray drying is also more and more considered. Indeed, this process is both continuous and more cost-efficient, as it utilizes less energy compared to freeze-drying; on the other hand, it exposes bacteria to higher heat and oxidative stresses. Apart from process optimization and strain selection, it is possible to enhance the resistance of bacteria by taking advantage of their adaptation capacity. Indeed, P. freudenreichii stress tolerance can be boosted by different pretreatments applied before the drying step, thus considerably increasing its final survival. In particular, adaptation to hyperosmotic conditions improves stress tolerance, while the presence of osmoprotectants may mitigate this improvement. Thermal adaptation also modulates tolerance towards these technological challenges. The composition of the growth medium, including the ratio between the carbohydrates provided and the non-protein nitrogen, plays a key role in driving the accumulation of osmoprotectants. This, in turn, determines P. freudenreichii tolerance towards different stresses, and overall towards both freeze-drying and spray-drying. As an example, the accumulation of trehalose enhances its spray-drying survival, while the accumulation of glycine betaine enhances its freeze-drying survival. Growth of propionibacteria in hyperconcentrated whey was used to trigger multiple stress tolerance acquisition, underpinned by overexpression of key stress protein, accumulation of cytoplasmic storage compounds, and leading to enhanced spray-drying survival. A simplified process, from cultivation to atomization, was developed by using whey as a 2-in-1 medium in which propionibacteria were grown, protected and dried with minimal cell death. This innovative process was then subjected to scaling up at the industrial level. In this aim, a gentle multi-stage drying process offering mild drying conditions by coupling spray drying with belt drying, led to final probiotic survival close to 100% when stress tolerance acquisition was previously implemented. Such innovation opens new avenues for the efficient, cost-effective and sustainable development of new probiotic production technologies, as well as probiotic application in the context of food and feed. KEY POINTS: • Propionibacteria acquire multi-stress tolerance when grown in hyper-concentrated whey. • Spray drying of osmo-adapted probiotic bacteria is possible with limited cell death. • A two-in-one drying method is developed to grow and dry probiotic bacteria in the same matrix.
丙酸杆菌是一种有益的食品级放线菌,广泛应用于各种食品中。作为主要应用,丙酸杆菌被用作奶酪成熟的起始剂,主要用于硬奶酪。事实上,在“瑞士型”奶酪(或开放式奶酪)的生产过程中,工艺过程有利于丙酸杆菌的生长和相应的丙酸发酵。这通过释放短链脂肪酸和通过脂解作用以及它们的特殊质地来赋予这些奶酪的特征风味。为了完成这种成熟,大量的丙酸杆菌被工业生产、干燥和储存,然后再用于奶酪制作。此外,丙酸杆菌以各种益生菌食品补充剂的形式商业化,旨在保持肠道健康和舒适,这与其产生有益代谢物(短链脂肪酸、维生素)以及免疫调节化合物的能力有关。丙酸杆菌的其他工业应用包括生产食品级 B 族维生素、海藻糖、共轭亚油酸和生物防腐剂。对于这些不同的应用,在生产、干燥、储存和最终实施过程中保持丙酸杆菌的存活和活性是至关重要的。更广泛地说,随着益生菌产品市场的增长,保持活的和有活性的益生菌细菌是一个挑战。大多数益生菌细菌都是冷冻干燥的,但喷雾干燥也越来越受到关注。事实上,与冷冻干燥相比,该过程不仅连续而且更具成本效益,因为它消耗的能量更少;另一方面,它使细菌暴露在更高的热量和氧化应激下。除了工艺优化和菌株选择外,还可以利用细菌的适应能力来提高其抗性。事实上,通过在干燥步骤之前进行不同的预处理,可以增强丙酸杆菌的耐受性,从而大大提高其最终存活率。特别是,适应高渗条件可以提高其耐受能力,而渗透压调节剂的存在可能会减轻这种改善。热适应也调节了对这些技术挑战的耐受能力。生长培养基的组成,包括提供的碳水化合物与非蛋白氮之间的比例,在驱动渗透压调节剂积累方面起着关键作用。这反过来又决定了丙酸杆菌对不同压力的耐受性,以及对冷冻干燥和喷雾干燥的总体耐受性。例如,海藻糖的积累增强了其喷雾干燥的生存能力,而甘氨酸甜菜碱的积累增强了其冷冻干燥的生存能力。丙酸杆菌在高浓度乳清中的生长被用来触发多种压力耐受性的获得,这是通过关键应激蛋白的过度表达、细胞质储存化合物的积累以及导致喷雾干燥生存能力增强来实现的。通过使用乳清作为 2-in-1 培养基,在其中生长、保护和干燥丙酸杆菌,同时最大限度地减少细胞死亡,开发了一种简化的从培养到雾化的工艺。然后,该创新工艺在工业规模上进行了放大。为此,通过将喷雾干燥与带式干燥相结合,采用温和的多阶段干燥工艺,提供温和的干燥条件,当以前实施了耐受能力获得时,最终益生菌的存活率接近 100%。这种创新为高效、经济高效和可持续发展新的益生菌生产技术以及益生菌在食品和饲料中的应用开辟了新的途径。关键点:• 丙酸杆菌在高浓度乳清中生长时会获得多种压力耐受性。• 喷雾干燥经过渗透压适应的益生菌细菌时,细胞死亡有限。• 开发了一种两用干燥方法,可在同一基质中生长和干燥益生菌细菌。