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使用新型B和C多物种引物通过聚合酶链反应筛选产γ-氨基丁酸的乳酸菌菌株以开发新型功能性食品

Selection of GABA-Producing Lactic Acid Bacteria Strains by Polymerase Chain Reaction Using Novel B and C Multispecies Primers for the Development of New Functional Foods.

作者信息

Langa Susana, Santos Silvia, Flores José Antonio, Peirotén Ángela, Rodríguez Susana, Curiel José Antonio, Landete José María

机构信息

Food Technology Department, National Institute for Agricultural and Food Research and Technology (INIA-CSIC), Carretera de La Coruña Km 7.5, 28040 Madrid, Spain.

出版信息

Int J Mol Sci. 2024 Dec 21;25(24):13696. doi: 10.3390/ijms252413696.

DOI:10.3390/ijms252413696
PMID:39769458
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11728273/
Abstract

Gamma-aminobutyric acid (GABA) has been attributed to health-promoting properties and has received attention from the food industry as an attractive bioactive compound for the development of functional foods. Some lactic acid bacteria (LAB) produce GABA through a glutamate decarboxylase encoded by B and a glutamate/GABA antiporter encoded by C. In this study, we develop a molecular screening method based on a polymerase chain reaction able to detect those genes in different LAB species through the use of novel multispecies primers. PCR was performed in 92 LAB strains of six different species. The primer pair designed for B allowed its identification in , , , , , and strains. For C, two different primer pairs were designed for its detection in different species. Glutamate decarboxylase activity (GAD assay) and GABase enzymatic quantification were also assessed. Among those strains showing glutamate decarboxylase activity, 93.2% harbored the B gene, and those showing GABA production had the B gene and exhibited glutamate decarboxylase activity. PCR detection of B correlates strongly with GABA production and constitutes a good strategy for the selection of LAB with high yields (>18 mM) that could be used for the development of GABA-enriched functional foods.

摘要

γ-氨基丁酸(GABA)具有促进健康的特性,作为一种有吸引力的生物活性化合物,已受到食品工业的关注,可用于开发功能性食品。一些乳酸菌(LAB)通过由B编码的谷氨酸脱羧酶和由C编码的谷氨酸/γ-氨基丁酸反向转运蛋白产生GABA。在本研究中,我们开发了一种基于聚合酶链反应的分子筛选方法,该方法能够通过使用新型多物种引物在不同的乳酸菌物种中检测这些基因。对六种不同物种的92株乳酸菌菌株进行了聚合酶链反应(PCR)。为检测B设计的引物对可在[具体六种菌的名称]菌株中鉴定出该基因。对于C,设计了两对不同的引物用于在不同物种中检测该基因。还评估了谷氨酸脱羧酶活性(GAD测定)和GABase酶定量。在那些显示谷氨酸脱羧酶活性的菌株中,93.2%含有B基因,而那些显示产生GABA的菌株含有B基因并表现出谷氨酸脱羧酶活性。对B的PCR检测与GABA的产生密切相关,是筛选高产(>18 mM)乳酸菌的良好策略,这些乳酸菌可用于开发富含GABA的功能性食品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/315b/11728273/9fedc9e330fd/ijms-25-13696-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/315b/11728273/9fedc9e330fd/ijms-25-13696-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/315b/11728273/9fedc9e330fd/ijms-25-13696-g001.jpg

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