Evaluation of Lipoxygenase Activity in Common Beans by UV and NMR Spectroscopy: Proposal for a Complementary Technique for Enzymatic Studies
Factors that reduce the quality of common bean during storage, including oxidative rancidity caused by lipoxygenase, have been extensively studied. In this context, this work evaluates lipoxygenase activity in five Carioca bean cultivars at different temperatures and storage times using ultraviolet (UV) and nuclear magnetic resonance (NMR) spectroscopies. UV absorption analyses demonstrated that the storage temperature of the grains influences the kinetic reaction rates during the enzymatic activity of lipoxygenase. Pinto beans and CNFC 10467 presented lower initial activity than the three BRS varieties but were more sensitive to high temperatures. BRS Estilo presented an anomalous behavior, with a strong reduction in activity at 15 and 21 °C until 72 days of storage. BRSMG Madrepérola and BRS Pontal presented stable enzymatic rates until 108 days of storage. A 26% reduction in fatty acid content, as a result of lipoxygenase action during storage, was observed both in solution-state and high resolution magic angle spinning (HR-MAS) NMR studies. These results highlight the importance of NMR in enzymatic kinetic reactions studies, complementing the results obtained by UV.
KeywordsPhaseolus vulgaris Oxidative enzyme Storage conditions Solution-state NMR 1H HR-MAS NMR
The authors are grateful to the Federal University of Goiás (UFG) for infrastructure support, as well as Financiadora de Estudos e Projetos (FINEP), Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for continued financial support and scholarships.
Embrapa for funding the project DarkBean. The Financier of Studies and Projects (FINEP) approved the purchase of NMR; the Foundation for Research Support of the State of Goiás (FAPEG) and the National Council for Scientific and Technological Development (CNPq) supported other projects, which indirectly bring resources to the laboratory. Thus, they are in Acknowledgments section.
Compliance with Ethical Standards
Conflict of Interest
Andressa Kuhnen Silva declares that he has no conflict of interest. Gerlon de Almeida Ribeiro Oliveira declares that he has no conflict of interest. Priscila Zaczuk Bassinello declares that she has no conflict of interest. Anna Cristina Lanna declares that she has no conflict of interest. Etelvino Henrique Novotny declares that he has no conflict of interest. Luciano Morais Lião declares that he has no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
- Araújo JMA (2008) Química de alimentos: teoria e prática, 4th edn. UFV, ViçosaGoogle Scholar
- Barison A, Silva CWP, Campos FR, Simonelli F, Lenz CA, Ferreira AG (2010) A simple methodology for the determination of fatty acid composition in edible oils through H NMR spectroscopy. Magn Reson Chem 48:642–650. https://doi.org/10.1002/mrc.2629
- Corrêa MM, de Carvalho LMJ, Nutti MR, de Carvalho JLV, Hohn Neto AR, Ribeiro EMG (2010) Water absorption, hard shell and cooking time of common beans (Phaseolus vulgaris L.). Afr J Food Sci Technol 1:13–20Google Scholar
- Hatumura PH, de Oliveira GS, Marcheafave GG, Rakocevic M, Bruns RE, Scarminio IS, Terrile AE (2018) Chemometric analysis of 1H NMR fingerprints of Coffea arabica green bean extracts cultivated under different planting densities. Food Anal Methods 11:1906–1914. https://doi.org/10.1007/s12161-017-1104-y CrossRefGoogle Scholar
- Kermasha S, Metche M (1986) Characterization of seed lipoxygenase of Phaseolus vulgaris cv, haricot. J Food Sci 51:1224–1227. https://doi.org/10.1111/j.1365-2621.1986.tb13090.x CrossRefGoogle Scholar
- Nelson DL, Cox MM (2005) Enzymes. In: Nelson DL, Cox MM (eds) Lehninger principles of biochemistry, 4th edn. W. H. Freeman and Company, New YorkGoogle Scholar
- Santos ONA, Folegatti MV, Dutra LM, Andrade IPS, Fanaya ED Jr, Lena BP, Barison A, Santos ADC (2017) Tracking lipid profiles of Jatropha curcas L. seeds under different pruning types and water managements by low-field and HR-MAS NMR spectroscopy. Ind Crop Prod 109:918–922. https://doi.org/10.1016/j.indcrop.2017.09.066 CrossRefGoogle Scholar
- Silva LMA, Alves Filho EG, Choze R, Liao LM, Alcantara GB (2012) 1H HRMAS NMR spectroscopy and Chemometrics for evaluation of metabolic changes in Citrus sinensis caused by Xanthomonas axonopodis pv. Citri. J Braz Chem Soc 23:1054–1061. https://doi.org/10.1590/S0103-50532012000600009 CrossRefGoogle Scholar
- Wasowicz E, Gramza A, Hęś M, Jeleń HH, Korczak J, Malecka M, Mildner-Szkudlarz S, Rudzińska M, Samotyja U, Zawirska-Wojtasiak R (2004) Oxidation of lipids in food. Pol J Food Nutr Sci 13/54(SI1):87–100Google Scholar