Purification and Characterization of Broccoli (<i>Brassica oleracea</i> var. <i>italica</i>) Myrosinase (β-Thioglucosidase Glucohydrolase)

Mahn, Andrea; Angulo, Alejandro; Cabañas, Fernanda

doi:10.1021/jf504957c

Cited by 24 publications

(38 citation statements)

References 25 publications

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“…Besides, since most authors use sinigrin in myrosinase studies, its use facilitates comparison with literature data. The highest specific activity of broccoli myrosinase and sinigrin was attained at pH 3.0, agreeing with Mahn et al [ 18 ], who reported that myrosinase activity was higher at acidic pH using sinigrin as substrate. Nevertheless, other authors reported that pH below 5 produced a structural destabilization of mustard seeds myrosinase.…”

Section: Discussionsupporting

confidence: 89%

“…Broccoli myrosinase has been poorly studied so far. This enzyme was purified for the first time by Mahn et al [ 18 ], and a preliminary characterization was reported. Recently, the cDNA nucleotide sequence of broccoli myrosinase was determined (Genbank ID: MF 461331); its amino acid sequence was deduced; and a three-dimensional model of its monomer was built (PMDB ID: 00811093) [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

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Molecular Docking of Potential Inhibitors of Broccoli Myrosinase

2018

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Glucosinolates are secondary metabolites occurring in Brassicaceae plants whose hydrolysis may yield isothiocyanates, widely recognized as health-promoting compounds. Myrosinase catalyzes this conversion. The chemical mechanism involves an unstable intermediary (thiohydroxamate-O-sulfonate) that spontaneously decomposes into isothiocyanates or other non-bioactive compounds depending on pH and cofactors. At acidic pH, non-bioactive compounds such as nitriles and thiocyanates are formed, while at neutral pH isothiocyanates are obtained. Broccoli myrosinase has been poorly studied so far. Recently, its amino acidic sequence was elucidated, and a structural model was built. The aim of this work was to study the molecular interaction of broccoli myrosinase with different ligands at acidic pH to propose possible inhibitors that prevent formation of undesirable compounds at acidic pH, and that at neutral pH dissociate from the enzyme, allowing formation of isothiocyanates. The interaction between broccoli myrosinase and 40 ligands was studied by molecular docking simulations. Both the enzyme and each inhibitor were set at pH 3.0. Amygdaline and arbutin showed the highest affinity to broccoli myrosinase in this condition. The residues that stabilize the complexes agree with those that stabilize the substrate (Gln207, Glu429, Tyr352, and Ser433). Accordingly, amygdaline and arbutin would perform as competitive inhibitors of myrosinase at pH 3.0.

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Section: Discussionsupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Molecular Docking of Potential Inhibitors of Broccoli Myrosinase

2018

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“…Broccoli ( Brassica oleracea ) is a popular vegetable because of its attractive green color and high nutritional value ( Martínez-Villaluenga et al, 2008 ). Many studies have focused on extracting bioactive metabolites from the edible parts of broccoli ( Assad et al, 2014 ; Mahn et al, 2014 ), extending the shelf life and maintaining the visual quality and content of bioactive compounds in broccoli florets ( Peng et al, 2015 ). However, the byproducts (in China about 200,000 tons of leaves and roots per year [ Su et al, 2005 ]) are left in the fields, leading not only to a waste of resources but also to a detrimental effect on the environment.…”

Section: Introductionmentioning

confidence: 99%

Effects of Replacement of Concentrate Mixture by Broccoli Byproducts on Lactating Performance in Dairy Cows

Yi¹,

Yang²,

Liu³

et al. 2015

Asian Australas. J. Anim. Sci

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The objective of the present study was to determine the effects of feeding pelletized broccoli byproducts (PBB) on milk yield and milk composition in dairy cows. In Trial 1, an in vitro gas test determined the optimal replacement level of PBB in a concentrate mixture in a mixed substrate with Chinese wild ryegrass hay (50:50, w/w) at levels of 0, 10%, 20%, 30%, or 40% (dry matter basis). When the concentrate was replaced by PBB at a level of 20%, no adverse effects were found on the gas volume or its rate constant during ruminal fermentation. In trial 2, 24 lactating cows (days in milk = 170.4±35; milk yield = 30±3 kg/d; body weight = 580 ±13 kg) were divided into 12 blocks based on day in milk and milk yield and randomly allocated to two dietary treatments: a basic diet with or without PBB replacing 20% of the concentrate mixture. The feeding trial lasted for 56 days; the first week allowed for adaptation to the diet. The milk composition was analyzed once a week. No significant difference in milk yield was observed between the two groups (23.5 vs 24.2 kg). A significant increase was found in milk fat content in the PBB group (p<0.05). Inclusion of PBB did not affect milk protein, lactose, total solids or solids-not-fat (p>0.05). These results indicated that PBB could be included in dairy cattle diets at a suitable level to replace concentrate mixture without any adverse effects on dairy performance.

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“…This finding is supported by a study where a lower bioavailability and delayed appearance of ITC metabolites were observed in the biological fluids (plasma and urine) of individuals consuming a broccoli supplement lacking myrosinase versus those consuming fresh broccoli sprouts. These differences may partially be explained by the fact that plant-and aphid-derived, but not bacterial-derived, myrosinases belong to the glycoside hydrolase (GH) family GH1 and exhibit differences in their activity [102,103].…”

Section: Anti-cancer Properties Of Gl Metabolites and Their Bioavailamentioning

confidence: 99%

Brassicaceae-Derived Anticancer Agents: Towards a Green Approach to Beat Cancer

Mandrich¹,

Caputo

2020

Nutrients

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Cancer is the main cause of mortality and morbidity worldwide. Although a large variety of therapeutic approaches have been developed and translated into clinical protocols, the toxic side effects of cancer treatments negatively impact patients, allowing cancer to grow. Brassica metabolites are emerging as new weapons for anti-cancer therapeutics. The beneficial role of the consumption of brassica vegetables, the most-used vegetables in the Mediterranean diet, particularly broccoli, in the prevention of chronic diseases, including cardiovascular diseases, diabetes, and obesity, has been well-documented. In this review, we discuss the anti-tumor effects of the bioactive compounds from Brassica vegetables with regard to the compounds and types of cancer against which they show activity, providing current knowledge on the anti-cancer effects of Brassica metabolites against major types of tumors. In addition, we discuss the impacts of industrial and domestic processing on the compounds’ functional properties before their consumption as well as the main strategies used to increase the content of health-promoting metabolites in Brassica plants through biofortification. Finally, the impacts of microbiota on the compounds’ bioactivity are considered. This information will be helpful for the further development of efficacious anti-cancer drugs.

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Purification and Characterization of Broccoli (Brassica oleracea var. italica) Myrosinase (β-Thioglucosidase Glucohydrolase)

Cited by 24 publications

References 25 publications

Molecular Docking of Potential Inhibitors of Broccoli Myrosinase

Molecular Docking of Potential Inhibitors of Broccoli Myrosinase

Effects of Replacement of Concentrate Mixture by Broccoli Byproducts on Lactating Performance in Dairy Cows

Brassicaceae-Derived Anticancer Agents: Towards a Green Approach to Beat Cancer

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