Identification and characterization of glutamate dehydrogenase activity in wild Lactococcus lactis isolated from raw milk cheeses

Cadiñanos, Luz P. Gómez de; Peláez, Carmen; Martínez-Cuesta, M. Carmen; García‐Cayuela, Tomás; Requena, Teresa

doi:10.1007/s00217-017-2988-x

Cited by 3 publications

(4 citation statements)

References 30 publications

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“…Aminotransferase assays are carried out with colorimetric L-glutamic acid assay kits (Gómez de Cadiñanos et al 2018). Reverse-phase HPLC has been used for the study of α-keto acids and aldehydes obtained by LAB enzymatic conversion of amino acids (Amarita et al 2006).…”

Section: Amino Acid Catabolismmentioning

confidence: 99%

Characterization of Lactic Acid Bacteria Used in Dairy Foods

Martínez-Cuesta¹,

García‐Cayuela²,

Peláez³

et al. 2021

Handbook of Dairy Foods Analysis

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This chapter covers methodologies currently in use for the selection and analysis of lactic acid bacteria (LAB) strains to be employed as starter cultures in dairy fermentations. The aspects covered are the methods for enumeration and identification of LAB strains and the evaluation of their technological properties for improving processing conditions and product quality during dairy fermentations. The selection of LAB strains based on health benefit properties and the conditions for the production and maintenance of dairy cultures are also addressed. The production of desirable and industrially robust starter dairy cultures can be achieved by choosing the desired phenotypic traits and the knowledge to select for these traits at the genotypic level. The analysis of a large number of strains for selecting specific properties requires high throughput screening tests to progressively reduce the number of candidates. The advances in omics technologies and analytical instruments have provided the bases to improve the evaluation and selection of LAB strains that are able to restore the unique characteristics of traditional dairy products and to benefit human health.

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Section: Amino Acid Catabolismmentioning

confidence: 99%

Characterization of Lactic Acid Bacteria Used in Dairy Foods

Martínez-Cuesta¹,

García‐Cayuela²,

Peláez³

et al. 2021

Handbook of Dairy Foods Analysis

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“…The quantification of gene expression by real-time RT-PCR was carried out on an iQ™5 Multicolor Real-Time PCR detection system Cycler (Bio-Rad) with IQ™ SYBR® Green Supermix (Bio-Rad) in 96-well plates. The primers were previously designed to target the araT and bcaT genes encoding aromatic and branched-chain amino acid specific aminotranferases, respectively, the kivD gene encoding an α-keto acid decarboxylase, the ytjE gene encoding a C-S lyase, the panE gene encoding a D-2hidroxyacid dehydrogenase and the gdh gene encoding the L. lactis glutamate dehydrogenase (García-Cayuela et al, 2012;Gómez de Cadiñanos et al, 2018). Primer annealing was set at 52 °C (30 s) and fluorescence was recorded during each extension step (72 °C, 60 s).…”

Section: Rna Extraction Reverse Transcription (Rt) and Quantification Of Gene Expression By Real-time Rt-pcrmentioning

confidence: 99%

“…The aromatic aminotransferase (AraT) and branched chain aminotransferase (BcaT) catalyze the transamination of aromatic and branchedchain amino acids, respectively (Yvon & Rijnen, 2001). The enzyme glutamate dehydrogenase (GDH) catalyzes the reversible conversion of glutamate into αketoglutarate (Gómez de Cadiñanos, Peláez, Martínez-Cuesta, García-Cayuela, & Requena, 2018;Tanous, Chambellon, Sepulchre, & Yvon, 2005). The α-keto acids formed by amino acid transamination play a central role in the biosynthetic and catabolic routes and are limiting factors for aroma formation (Yvon & Rijnen, 2001).…”

Section: Introductionmentioning

confidence: 99%

“…Later, we studied the effect of BCAA content on the growth of L. lactis IFPL730 and we observed that the expression of some functional genes related to amino acid catabolism (kivD, bcaT and ytjE) increased markedly when growing under both isoleucine and valine starvation conditions (García-Cayuela, Gómez de Cadiñanos, Peláez, & Requena, 2012). Aminotransferase activities are widespread among L. lactis, whereas GDH and KivD are key aroma enzymes that show low prevalence and great strain dependence (De la Plaza et al, 2006;Gómez de Cadiñanos et al, 2018). L. lactis IFPL953 is a wild strain isolated from raw milk cheese characterized by a wide enzymatic background for aroma production, including AraT and BcaT activities, the GDH activity to produce αketoglutarate required for transamination reactions, and the KivD and C-S lyase (YtjE) activities.…”

Section: Introductionmentioning

confidence: 99%

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Expression of amino acid converting enzymes and production of volatile compounds by Lactococcus lactis IFPL953

Cadiñanos

García‐Cayuela

Martínez-Cuesta

et al. 2019

International Dairy Journal

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Lactococcus lactis contains a large number of key enzymes responsible for the formation of volatile compounds characteristic of cheese flavour. In the present work we have investigated the expression of genes codifying amino acid converting enzymes (AACE) and the formation of volatile compounds by L. lactis IFPL953 and its mutant lacking the hydroxy acid dehydrogenase PanE (L. lactis IFPL953ΔpanE). The growth in absence of isoleucine was the main induction factor in the expression of araT, bcaT, panE, and kivD. The expression of the gdh gene of L. lactis IFPL953ΔpanE increased remarkably during the stationary growth phase, particularly under isoleucine and valine starvation conditions. L. lactis IFPL953ΔpanE showed an enhanced formation of 2-and 3-methylbutanal and their corresponding alcohols and an increase in the formation of ketones. These findings contribute to a better knowledge of L. lactis AACE regulation and its potential application in cheese flavour formation.

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Pyruvate stimulates transamination of leucine into α-ketoisocaproic acid and supports 3-methylbutanal production by Lactococcus lactis

Brandsma,

Brinkman,

Wolkers-Rooijackers

et al. 2024

Journal of Applied Microbiology

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Aim To investigate the effect of pyruvate and glucose on leucine transamination and 3-methylbutanal production by Lactococcus lactis, including the comparison with cells possessing glutamate dehydrogenase (GDH) activity. Methods and Results L. lactis cells were incubated in chemically defined medium (CDM) with the pH controlled at 5.2 to mimic cheese conditions. Pyruvate supplementation stimulated the production of the key flavour compound 3-methylbutanal by 3 to 4 times after 72 hours of incubation. Concurrently, alanine production increased, demonstrating the involvement of pyruvate in transamination reactions. Glucose-metabolising cells excreted α-ketoisocaproic acid and produced even 3 times more 3-methylbutanal after 24 hours than pyruvate-supplemented cells. Conjugal transfer technique was used to transfer the plasmid pGdh442 carrying the gdh gene encoding for GDH to L. lactis. Introducing GDH did not stimulate the excretion of α-ketoisocaproic acid and the production of 3-methylbutanal. Conclusions These results demonstrate that Lactococcus uses pyruvate to transaminate leucine into α-ketoisocaproic acid which supports 3-methylbutanal production. Surprisingly, GDH activity did not stimulate leucine transamination and 3-methylbutanal production.

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Identification and characterization of glutamate dehydrogenase activity in wild Lactococcus lactis isolated from raw milk cheeses

Cited by 3 publications

References 30 publications

Characterization of Lactic Acid Bacteria Used in Dairy Foods

Characterization of Lactic Acid Bacteria Used in Dairy Foods

Expression of amino acid converting enzymes and production of volatile compounds by Lactococcus lactis IFPL953

Pyruvate stimulates transamination of leucine into α-ketoisocaproic acid and supports 3-methylbutanal production by Lactococcus lactis

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