Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sake

Lactobacillus acetotolerans and Fructilactobacillus fructivorans are responsible for the spoilage of Japanese sake, and sake brewers want to control these bacteria to prevent product-quality deterioration. Due to these bacteria's slow growth, a long time must pass before a product's spoila...

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Main Authors: Masayuki Takahashi, Kana Morikawa, Takeshi Akao
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Applied Food Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S2772502225001453
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author Masayuki Takahashi
Kana Morikawa
Takeshi Akao
author_facet Masayuki Takahashi
Kana Morikawa
Takeshi Akao
author_sort Masayuki Takahashi
collection DOAJ
description Lactobacillus acetotolerans and Fructilactobacillus fructivorans are responsible for the spoilage of Japanese sake, and sake brewers want to control these bacteria to prevent product-quality deterioration. Due to these bacteria's slow growth, a long time must pass before a product's spoilage can be identified by a culture-dependent method; a method enabling the rapid detection of spoilage is thus desired for maintaining sake's quality and reducing processing times. Herein, we developed a novel, simple method for determining the ori copy and ter copy numbers of both bacterial species by a single multiplex digital polymerase chain reaction (PCR). Our investigation about the ori and ter copy numbers of L. acetotolerans RIB9124 strain revealed that (i) the ter-region copy number was highly correlated with the colony-forming units, and (ii) the ori:ter ratio during log phase (mean: 1.36) was higher than that during non-log phase (mean: 1.16). The relationship between the ori:ter ratio and the bacterial doubling time was parabola-shaped, not linear. We constructed a model for estimating the two species' doubling times based on the ori:ter ratio and then created a generalized linear model with a probit link function using the ori:ter ratio, the doubling time predicted by the ori:ter ratio, and the ter copy number as parameters. The model was able to predict whether sake inoculated with either species would spoil within 4 months (spoilage-prediction error rate validated with 45 samples: 15.6 %). In light of these results, we suggest some criteria for judging the risk of Japanese sake spoiling within 4 months.
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spelling doaj-art-5b1a826f97be4364a01c087dd33e4ad22025-08-20T03:45:15ZengElsevierApplied Food Research2772-50222025-06-015110083510.1016/j.afres.2025.100835Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sakeMasayuki Takahashi0Kana Morikawa1Takeshi Akao2Correspondence author at. National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan.; National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, JapanNational Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, JapanNational Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, JapanLactobacillus acetotolerans and Fructilactobacillus fructivorans are responsible for the spoilage of Japanese sake, and sake brewers want to control these bacteria to prevent product-quality deterioration. Due to these bacteria's slow growth, a long time must pass before a product's spoilage can be identified by a culture-dependent method; a method enabling the rapid detection of spoilage is thus desired for maintaining sake's quality and reducing processing times. Herein, we developed a novel, simple method for determining the ori copy and ter copy numbers of both bacterial species by a single multiplex digital polymerase chain reaction (PCR). Our investigation about the ori and ter copy numbers of L. acetotolerans RIB9124 strain revealed that (i) the ter-region copy number was highly correlated with the colony-forming units, and (ii) the ori:ter ratio during log phase (mean: 1.36) was higher than that during non-log phase (mean: 1.16). The relationship between the ori:ter ratio and the bacterial doubling time was parabola-shaped, not linear. We constructed a model for estimating the two species' doubling times based on the ori:ter ratio and then created a generalized linear model with a probit link function using the ori:ter ratio, the doubling time predicted by the ori:ter ratio, and the ter copy number as parameters. The model was able to predict whether sake inoculated with either species would spoil within 4 months (spoilage-prediction error rate validated with 45 samples: 15.6 %). In light of these results, we suggest some criteria for judging the risk of Japanese sake spoiling within 4 months.http://www.sciencedirect.com/science/article/pii/S2772502225001453Lactobacillus acetotoleransFructilactobacillus fructivoransHiochiSakeSpoilageOri
spellingShingle Masayuki Takahashi
Kana Morikawa
Takeshi Akao
Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sake
Applied Food Research
Lactobacillus acetotolerans
Fructilactobacillus fructivorans
Hiochi
Sake
Spoilage
Ori
title Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sake
title_full Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sake
title_fullStr Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sake
title_full_unstemmed Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sake
title_short Novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of Japanese sake
title_sort novel method for predicting the risk of spoilage by lactic acid bacteria during the storage of japanese sake
topic Lactobacillus acetotolerans
Fructilactobacillus fructivorans
Hiochi
Sake
Spoilage
Ori
url http://www.sciencedirect.com/science/article/pii/S2772502225001453
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