Domingues, MR

Fontes, Ana Luiza

Gomes, Ana Maria

Pimentel, Lígia

Rodriguez, Juan Miguel

Rodríguez-Alcalá, Luís M.

Salsinha, Ana Sofia

English

Fontes, Ana L.

Pimentel, Lígia L.

Salsinha, Ana S.

Rodríguez, Juan M.

Domingues, M. Rosário

Rodríguez-Alcalá, Luis M.

Gomes, Ana M.

Repositório Institucional da Universidade Católica Portuguesa

1Universidade Católica Portuguesa, CBQF - Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Porto, Portugal2QOPNA – Unidade de Investigação de Química Orgânica, Produtos Naturais e Agroalimentares, Universidade de Aveiro, Aveiro, Portugal3CINTESIS – Centro de Investigação em Tecnologias e Sistemas de Informação em Saúde, Faculdade de Medicina da Universidade do Porto, Porto, Portugal4Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, Madrid, Spain5Mass Spectrometry Centre, Department of Chemistry & QOPNA, University of Aveiro, Campus Universitário de Santiago Aveiro, Portugal6Department of Chemistry & CESAM & ECOMARE, University of Aveiro, Campus Universitário de Santiago, Aveiro, PortugalInfluence of shaking and viable cell numbers on microbial conjugated linoleic acid (CLA) productionAna L. Fontes1,2, Lígia L. Pimentel1,2,3, Ana S. Salsinha1, Juan M. Rodríguez4, M. Rosário Domingues5,6, Luis M. Rodríguez-Alcalá1 and Ana M. Gomes1IntroductionMethodsConclusionsAcknowledgementsThis work was supported by National Funds from FCT – Fundação para a Ciência e Tecnologia through project “Pro-TECh-CLnA - Microbial Production ofBioactive Conjugated Linolenic Acid Isomers to Obtain Functional Ingredients and Foods” reference PTDC/AGR-TEC/2125/2014. We would also like to thankthe scientific collaboration under the FCT project UID/Multi/50016/2019. Financial support for the authors A.L. Fontes and L.L. Pimentel was provided byfellowships SFRH/BD/117721/2016 and SFRH/BPD/119785/2016, respectively, granted by the Portuguese government through FCT.Conjugated linoleic acid (CLA) isomers have been attracting much scientific attention due to their potential bioactive properties1. These compounds are naturallyproduced during the biohydrogenation process of dietary linoleic acid (LA) by ruminal bacteria, however, several probiotic strains isolated from human intestineand dairy products have also revealed the capacity to produce those bioactive fatty acids2,3. Some studies have reported that the amount of CLA produced isstrain-dependent and may also be influenced by cultivation conditions4,5. In vitro studies usually test CLA production by growing strains in culture mediumcontaining a specific LA concentration6, but few have applied shaking during incubation. Shaking may improve access to substrate and result in better CLA yields,therefore, this work aimed to study the influence of shaking on CLA production by strains previously identified as CLA-producers.In conclusion, shaking and viable cell numbers (in agreement of incubation time) affect microbial CLA production in a strain-dependent manner.ResultsShaking reduced LA conversion rates for all strains except for L. gasseri VR-III-51 (Figure 1).In what concerns correlation between viable cell numbers (data not shown)at the end of each incubation period and LA conversion rates, it wasobserved a negative correlation for L. gasseri VR-III-51 with shaking, apositive correlation for B. breve DSM 20091 without shaking (Figure 2) and nocorrelation for L. plantarum 20205 and L. sakei DSM 20017, either with orwithout shaking.References1. Yang B, Chen H, Stanton C, et al. Review of the roles of conjugated linoleic acid in health and disease. J Funct Foods. 2015;15:314-325.2. Shokryzadan P, Rajion MA, Meng GY, et al. Conjugated linoleic acid: a potent fatty acid linked to animal and human health. Crit Rev Food Sci Nutr. 2017;57(13):2737-2748.3. Fontes AL, Pimentel LL, Simões CD, Gomes AMP, Rodríguez-Alcalá LM. Evidences and perspectives in the utilization of CLNA isomers as bioactive compounds in foods. Crit Rev Food Sci Nutr. 2017;57(12):2611-2622.4. Renes E, Linares DM, González L, Fresno JM, Tornadijo ME, Stanton C. Production of conjugated linoleic acid and gamma-aminobutyric acid by autochthonous lactic acid bacteria and detection of the genes involved. J Funct Foods. 2017;34:340-346.5. Terán V, Pizarro PL, Zacarías MF, Vinderola G, Medina R, Van Nieuwenhove C. Production of conjugated dienoic and trienoic fatty acids by lactic acid bacteria and bifidobacteria. J Funct Foods. 2015;19:417-425.6. Yang B, Gao H, Stanton C, et al. Bacterial conjugated linoleic acid production and their applications. Prog Lipid Res. 2017;68:26-36.7. Pimentel LL, Fontes AL, Gomes AM, Rodríguez-Alcalá LM. Considerations about the in situ derivatization and fractionation of EFA and NEFA in biological and food samples. MethodsX. 2015;2:475-484.Bifidobacterium breve DSM 20091Lactobacillus gasseri VR-III-51Lactobacillus plantarum DSM 20205Lactobacillus sakei DSM 200171% (v/v)MRS + 0,5 mg/mL LA37 ⁰C; 24, 48 and 72 h; 150 rpm 37 ⁰C; 24, 48 and 72 h; no shakingFatty acid analysis through GC-FIDViable cell counts in MRS agar platesSupernatant treatement according to Pimentel et al.7Serial dilutionsACBDFigure 1. LA conversion rates of B. breve DSM 20091 (A), L. gasseri VR-III-51 (B), L. plantarum DSM 20205 (C) and L. sakei DSM 20017 (D) with (orange) and without (blue) shaking. Different superscript letters for significant differences (p<0,05) between shaking condition.A BFigure 2. Correlation between LA conversion rate and viable cell numbers of L. gasseri VR-III-51 with shaking (A) and B. breve DSM 20091 without shaking (B). P<0,05 according to Kendall correlation test. 

Influence of shaking and viable cell numbers on microbial conjugated linoleic acid (CLA) production

7-11 July 2019 | Glasgow, Scotland | www.fems2019.org8th Congress of European MicrobiologistsABSTRACT BOOKIn collaboration with1560  PW424 Influence of shaking and viable cell numbers on microbial conjugated linoleic acid (CLA) production  Ana Luiza Fontes1;2, Lígia Pimentel1;2;3, Ana Sofia Salsinha1, Juan Miguel Rodriguez4, MR Domingues5;6, Luís M. Rodríguez-Alcalá1, Ana Maria Gomes1  1Universidade Católica Portuguesa, CBQF - Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia 2QOPNA – Unidade de Investigação de Química Orgânica, Produtos Naturais e Agroalimentares, Universidade de Aveiro 3CINTESIS – Centro de Investigação em Tecnologias e Sistemas de Informação em Saúde, Faculdade de Medicina da Universidade do Porto 4Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid 5Mass Spectrometry Centre, Department of Chemistry & QOPNA, Universidade de Aveiro, Campus Universitário de Santiago 6Department of Chemistry & CESAM & ECOMARE, Universidade de Aveiro, Campus Universitário de Santiago   Background: Conjugated linoleic acid (CLA) isomers are naturally produced from dietary linoleic acid (LA) by ruminal bacteria, but several probiotic strains have also revealed the capacity to produce these bioactive compounds. The amount of CLA produced is strain-dependent and may also be influenced by cultivation conditions. Objectives: In vitro studies usually test CLA production by growing strains in culture medium containing a specific LA concentration, but few have applied shaking during incubation; shaking may improve access to substrate. Hence, this work aimed to study the influence of shaking on CLA production. Methods: Bifidobacterium breve DSM 20091, Lactobacillus gasseri VR-III-51, L. plantarum DSM 20205 and L. sakei 20017, previously identified as CLA-producers, were selected for this study. They were inoculated (1% v/v) in MRS medium, containing 0.5 mg/mL of LA and incubated at 37 °C for either 24, 48 or 72 h, with and without shaking (150 rpm). Viable cell numbers were determined and fatty acid concentration was analyzed by gas chromatography. Results: Shaking reduced LA conversion rates for all strains except L. gasseri VR-III-51. In what concerns correlation between viable cell numbers at the end of each incubation period and LA conversion rates, important differences were observed: a negative correlation for L. gasseri VR-III-51 with shaking, a positive correlation for B. breve DSM 20091 without shaking and no correlation for L. plantarum 20205 and L. sakei DSM 20017, with/without shaking were observed. In conclusion, shaking and viable cell numbers (incubation time) affect microbial CLA production in a strain-dependent manner.  

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Influence of shaking and viable cell numbers on microbial conjugated linoleic acid (CLA) production

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Repositório Institucional da Universidade Católica Portuguesa

Repositório Institucional da Universidade Católica Portuguesa