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优化培养基培养的乳酸菌对碳黑曲霉生长和赭曲霉毒素 A 产生的抑制作用。

Inhibition of Aspergillus carbonarius growth and Ochratoxin A production using lactic acid bacteria cultivated in an optimized medium.

机构信息

Department of Biotechnology, Metropolitan Autonomous University, ZC, 09310, Iztapalapa, Mexico City, Mexico.

Aix Marseille Université, Avignon Université, CNRS, IRD, IMBE, Marseille, France.

出版信息

Int J Food Microbiol. 2023 Nov 2;404:110320. doi: 10.1016/j.ijfoodmicro.2023.110320. Epub 2023 Jul 18.

Abstract

The Gram-positive bacteria lactic acid bacteria (LAB) are used in the food industry but are also known for inhibiting certain food spoilage microorganisms, especially fungi. Sources of nitrogen (N) for culture media are generally organic and expensive. Many attempts have been made to formulate economical culture media with alternative N sources obtained from agricultural and industrial byproducts. This study describes the design and optimization of an inexpensive culture medium for Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) MZ809351 strain B31. The culture medium was optimized using statistical experimental designs to identify the factors with the most significant effects on biomass concentration to reduce the overall cost, aiming to obtain a biomass concentration similar to that obtained with the reference LAB culture medium (de Man, Rogosa and Sharpe; MRS). Sodium acetate and magnesium sulfate were the most significant factors (p < 0.005), and their contents were reduced by 22 % and 40 %, respectively, without affecting biomass concentration. Malt germ extract (MGE) was used as an alternative nitrogen source to replace meat extract (ME) and proteose peptone (PP). Through these experiments, the composition of a culture medium that is less expensive than MRS broth was defined, which produced a biomass concentration (3.8 g/L) similar to that obtained with MRS medium. The inhibitory effects of two LAB strains isolated from the Ivory Coast and Mexico on the growth and production of ochratoxin A (OTA) in an ochratoxigenic fungus was tested. The minimum cellular concentration of the LAB to prevent the development of Aspergillus carbonarius Ac 089 and the production of OTA was determined in a model assay in Petri dishes. The conditions to inhibit the germination of A. carbonarius Ac 089 and the production of OTA were found. Using the optimized medium and a ratio of 2 × 10 LAB/spore (1 × 10 CFU/mL) strain B7 (L. plantarum MZ809351) and 2 × 10 LAB/spore (1 × 10 CFU/mL) strain B31 (L. plantarum MN922335) completely inhibited the growth of the fungus. A ratio of 2 × 10 LAB/spore (1 × 10 CFU/mL) was required to inhibit OTA production with strains B7 and B31. This study indicates the potential of cultivating LAB in an optimized and inexpensive culture medium for use as a biological control agent against ochratoxigenic fungi in food.

摘要

革兰氏阳性细菌乳酸菌(LAB)在食品工业中被广泛应用,同时还能抑制某些食品腐败微生物,尤其是真菌的生长。培养基中的氮(N)源通常是有机的且昂贵的。为了寻找替代的 N 源,许多人尝试使用来自农业和工业副产品的经济型培养基来配制培养基。本研究描述了一种用于植物乳杆菌(原为植物乳杆菌)MZ809351 菌株 B31 的经济型培养基的设计和优化。使用统计实验设计来确定对生物量浓度有显著影响的因素,以降低总成本,从而获得与参考 LAB 培养基(de Man、Rogosa 和 Sharpe;MRS)相似的生物量浓度。乙酸钠和硫酸镁是最显著的因素(p < 0.005),其含量分别降低了 22%和 40%,而生物量浓度不受影响。麦芽提取物(MGE)被用作替代氮源,以替代肉提取物(ME)和蛋白胨(PP)。通过这些实验,定义了一种比 MRS 肉汤更便宜的培养基组成,该培养基产生的生物量浓度(3.8 g/L)与 MRS 培养基相似。测试了从科特迪瓦和墨西哥分离的两种 LAB 菌株对产赭曲霉毒素 A(OTA)的真菌生长和产毒的抑制作用。在平板模型试验中,确定了 LAB 细胞的最低浓度,以防止烟曲霉 Ac 089 的生长和 OTA 的产生。找到了抑制烟曲霉 Ac 089 发芽和 OTA 产生的条件。使用优化的培养基和 2×10LAB/spore(1×10CFU/mL)B7 株(植物乳杆菌 MZ809351)和 2×10LAB/spore(1×10CFU/mL)B31 株(植物乳杆菌 MN922335)的比例,完全抑制了真菌的生长。B7 和 B31 菌株抑制 OTA 产生需要 2×10LAB/spore(1×10CFU/mL)的比例。本研究表明,在优化和廉价的培养基中培养 LAB 作为食品中产赭曲霉毒素真菌的生物防治剂具有潜力。

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