A Species-Specific qPCR Method for Enumeration of Lactobacillus sanfranciscensis , Lactobacillus brevis , and Lactobacil
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A Species-Specific qPCR Method for Enumeration of Lactobacillus sanfranciscensis, Lactobacillus brevis, and Lactobacillus curvatus During Cocultivation in Sourdough Hyun-Wook Baek 1 & Seul-Ah Kim 2 & Won-Ki Min 3 & Shin Dal Kang 4 & Sangmin Shim 4 & Nam Soo Han 2
&
Jin-Ho Seo 1,4
Received: 12 July 2020 / Accepted: 15 November 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Lactobacillus sanfranciscensis, Lactobacillus brevis, and Lactobacillus curvatus are frequently isolated from cereal-based fermented foods and are the dominant lactic acid bacteria in Korean sourdough. Detection of the individual species and their enumeration during fermentation are of great importance for maintaining the quality of bakery products. Here, we developed a species-specific quantitative polymerase chain reaction (qPCR) method to monitor the population of these three bacterial species during cocultivation in sourdough. Target genes, including those encoding nucleoside hydrolase, hypothetical protein, and glyoxalase, were selected by MegaBLAST for the detection of L. sanfranciscensis, L. brevis, and L. curvatus, respectively. The specificities of PCR primer sets were verified with qPCR; constant cycle threshold (Ct) values were obtained from mixed genomic DNAs (gDNAs) of target and non-target strains. The qPCR results were unaffected by the sourdough matrix. The cell numbers derived from qPCR were 10–65% higher than those obtained using a conventional plate-counting method. The qPCR standard curves for the three Lactobacillus species were established, and their populations were successfully enumerated during sourdough propagation. This method would enable quantification of three Lactobacillus species during cocultivation in sourdough and provide useful information on microbial commensalism that is essential to obtain high-quality bread. Keywords Sourdough . Real-time quantitative polymerase chain reaction . Lactobacillus sanfranciscensis . Lactobacillus brevis . Lactobacillus curvatus
Introduction Sourdough has been used in bread production worldwide due to the improved rheological, organoleptic, and nutritional properties compared to the products made by straight dough Hyun-Wook Baek and Seul-Ah Kim contributed equally to this work. * Nam Soo Han [email protected] 1
Department of Agricultural Biotechnology, Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea
2
Brain Korea 21 Center for Bio-Resource Development, Division of Animal, Horticultural, and Food Sciences, Chungbuk National University, Cheongju 28644, Republic of Korea
3
Department of Food Science and Development, Kyungil University, Gyeongbuk 38428, Republic of Korea
4
Research Institute of Food and Biotechnology, SPC Group, Seoul 08826, Republic of Korea
processes (Hansen and Schieberle 2005; Pétel et al. 2017). The difference between two bakery products is based on metabolic compounds produced by lactic acid bacteria (LAB) at dough stage (Gänzle and Ripari 2016). The beneficial contrib
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