Isolation and purification of lactic acid bacteria and yeasts based on a multi-channel magnetic flow device and rapid qualitative and quantitative detection

Huilin Wu , Shijie Liu , Miaoyun Li, Lijun Zhao, Yaodi Zhu, Gaiming Zhao, Yangyang Ma, Lingxia Sun, Yanxia Liu, Dong Liang
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Abstract

Rapid isolation and identification of lactic acid bacteria and yeasts during fermentation is of great significance for quality control and regulation of fermented foods. In this study, we prepared a multi-channel magnetic flow device for rapid separation and purification of lactic acid bacteria and yeast, and based on SERS spectrum, we made rapid qualitative and quantitative analysis of Lactobacillus plantarum, Lactococcus lactis and Saccharomyces cerevisiae. The results showed that the synthesized Synthesized Fe3O4-Van antibiotic magnetic beads are paramagnetic; Fe3O4-Van antibiotic magnetic beads achieved capture efficiencies of more than 98.5 % for both L. plantarum and L. lactis at 102–104 CFU/mL, respectively. Separation and purification efficiency of single S. cerevisiae, L. plantarum and L. lactis by multi-channel magnetic flow device all reached more than 98 % with good isolation and purification results. The SERS spectra of the three microorganisms were classified and analyzed using linear discriminant analysis (LDA), and the accuracy of the established LDA model was 100 %, which completely differentiated the SERS spectra of the three microorganisms,and realized the qualitative identification of L. plantarum, L. lactis, and S. cerevisiae, and finally, quantitative model was established with the logarithmic values (lg C) of different concentrations of L. plantarum, L. lactis, and S. cerevisiae as the horizontal coordinates, and the Raman intensities at their strongest characteristic peaks of 512 cm−1, 1669 cm−1, and 1125 cm−1, respectively, were used as vertical coordinates to establish a quantitative model, with the lowest detection limit of 10 CFU/mL, and the digital quantification of lactic acid bacteria and yeast were achieved. It provided an effective means for real-time monitoring and tracking of the dynamics of lactic acid bacteria and yeast in the fermentation process and the quality control of fermented foods.

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基于多通道磁流装置的乳酸菌和酵母的分离和纯化以及快速定性和定量检测。
快速分离和鉴定发酵过程中的乳酸菌和酵母菌对发酵食品的质量控制和监管具有重要意义。本研究制备了一种用于快速分离纯化乳酸菌和酵母菌的多通道磁流装置,并基于 SERS 光谱对植物乳杆菌、乳酸乳球菌和酿酒酵母进行了快速定性和定量分析。结果表明,合成的 Fe3O4-Van 抗生素磁珠具有顺磁性;Fe3O4-Van 抗生素磁珠对植物乳杆菌和乳球菌的捕获效率均超过 98.5%,捕获量分别为 102-104 CFU/mL。利用多通道磁流装置分离纯化单个 S. cerevisiae、L. plantarum 和 L. lactis 的效率均达到 98 % 以上,分离纯化效果良好。利用线性判别分析(LDA)对三种微生物的 SERS 光谱进行了分类和分析,建立的 LDA 模型的准确率为 100%,完全区分了三种微生物的 SERS 光谱,实现了对植物乳杆菌、乳酸菌和麦角菌的定性鉴定,最后利用不同浓度的植物乳杆菌、乳酸菌和麦角菌的对数值(lg C)建立了定量模型。以植物乳杆菌、乳酸菌和酵母菌的对数值(lg C)为横坐标,分别以其最强特征峰 512 cm-1、1669 cm-1 和 1125 cm-1 的拉曼强度为纵坐标建立定量模型,最低检测限为 10 CFU/mL,实现了乳酸菌和酵母菌的数字化定量。它为实时监测和跟踪发酵过程中乳酸菌和酵母的动态以及发酵食品的质量控制提供了有效手段。
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来源期刊
CiteScore
8.40
自引率
11.40%
发文量
1364
审稿时长
40 days
期刊介绍: Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy (SAA) is an interdisciplinary journal which spans from basic to applied aspects of optical spectroscopy in chemistry, medicine, biology, and materials science. The journal publishes original scientific papers that feature high-quality spectroscopic data and analysis. From the broad range of optical spectroscopies, the emphasis is on electronic, vibrational or rotational spectra of molecules, rather than on spectroscopy based on magnetic moments. Criteria for publication in SAA are novelty, uniqueness, and outstanding quality. Routine applications of spectroscopic techniques and computational methods are not appropriate. Topics of particular interest of Spectrochimica Acta Part A include, but are not limited to: Spectroscopy and dynamics of bioanalytical, biomedical, environmental, and atmospheric sciences, Novel experimental techniques or instrumentation for molecular spectroscopy, Novel theoretical and computational methods, Novel applications in photochemistry and photobiology, Novel interpretational approaches as well as advances in data analysis based on electronic or vibrational spectroscopy.
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