Food Microbiology, Wageningen University and Research, PO Box 17, 6700 AA, Wageningen, The Netherlands.
BU Veterinary Drugs, RIKILT, Wageningen University and Research, Akkermaalsbos 2, 6708 WB, Wageningen, The Netherlands.
Microb Cell Fact. 2019 Aug 6;18(1):129. doi: 10.1186/s12934-019-1179-9.
Vitamin K2 (menaquinone, MK-n) is a lipid-soluble vitamin that functions as a carboxylase co-factor for maturation of proteins involved in many vital physiological processes in humans. Notably, long-chain vitamin K2 is produced by bacteria, including some species and strains belonging to the group of lactic acid bacteria (LAB) that play important roles in food fermentation processes. This study was performed to gain insights into the natural long-chain vitamin K2 production capacity of LAB and the factors influencing vitamin K2 production during cultivation, providing a basis for biotechnological production of vitamin K2 and in situ fortification of this vitamin in food products.
We observed that six selected Lactococcus lactis strains produced MK-5 to MK-10, with MK-8 and MK-9 as the major MK variant. Significant diversities between strains were observed in terms of specific concentrations and titres of vitamin K2. L. lactis ssp. cremoris MG1363 was selected for more detailed studies of the impact of selected carbon sources tested under different growth conditions [i.e. static fermentation (oxygen absent, heme absent); aerobic fermentation (oxygen present, heme absent) and aerobic respiration (oxygen present, heme present)] on vitamin K2 production in M17 media. Aerobic fermentation with fructose as a carbon source resulted in the highest specific concentration of vitamin K2: 3.7-fold increase compared to static fermentation with glucose, whereas aerobic respiration with trehalose resulted in the highest titre: 5.2-fold increase compared to static fermentation with glucose. When the same strain was applied to quark fermentation, we consistently observed that altered carbon source (fructose) and aerobic cultivation of the pre-culture resulted in efficient vitamin K2 fortification in the quark product.
With this study we demonstrate that certain LAB strains can be employed for efficient production of long-chain vitamin K2. Strain selection and optimisation of growth conditions offer a viable strategy towards natural vitamin K2 enrichment of fermented foods, and to improved biotechnological vitamin K2 production processes.
维生素 K2(甲萘醌,MK-n)是一种脂溶性维生素,作为羧化酶辅助因子,参与人体许多重要生理过程中蛋白质的成熟。值得注意的是,长链维生素 K2 由细菌产生,包括一些属于乳酸菌(LAB)的物种和菌株,它们在食品发酵过程中发挥着重要作用。本研究旨在深入了解 LAB 天然长链维生素 K2 的生产能力,以及培养过程中影响维生素 K2 生产的因素,为维生素 K2 的生物技术生产和食品中该维生素的原位强化提供依据。
我们观察到,六种选定的乳球菌菌株产生了 MK-5 至 MK-10,其中 MK-8 和 MK-9 是主要的 MK 变体。菌株之间在维生素 K2 的特定浓度和效价方面存在显著差异。选择乳球菌亚种乳脂乳球菌 MG1363 进行更详细的研究,研究了在不同生长条件[即静态发酵(缺氧,无血红素);有氧发酵(缺氧,无血红素)和有氧呼吸(含氧,有血红素)]下,测试的选定碳源对 M17 培养基中维生素 K2 生产的影响。以果糖作为碳源进行有氧发酵,可使维生素 K2 的比浓度增加 3.7 倍,与以葡萄糖作为碳源进行静态发酵相比;而以海藻糖作为碳源进行有氧呼吸,可使维生素 K2 的效价增加 5.2 倍,与以葡萄糖作为碳源进行静态发酵相比。当同一菌株应用于酸奶发酵时,我们始终观察到,改变碳源(果糖)和预培养的有氧培养可有效地强化酸奶产品中的维生素 K2。
本研究表明,某些 LAB 菌株可用于高效生产长链维生素 K2。菌株选择和优化生长条件为发酵食品中天然维生素 K2 的富集以及改进生物技术生产维生素 K2 的工艺提供了可行的策略。