A fast and gentle method for the isolation of myrosinase complexes from Brassicaceous seeds

Bellostas, Natalia; Petersen, Iben Lykke; Sørensen, Jens Christian; Sørensen, Hilmer

doi:10.1016/j.jprot.2007.11.006

Cited by 26 publications

(31 citation statements)

References 39 publications

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“…It is, anyway, more likely that the observed degradation of the glucosinolates is a result of a fast interaction with the active enzyme myrosinase, as shown in previous studies (Bellostas et al, 2008). The pattern observed in glucosinolate concentration related to pressure is similar to the results obtained when broccoli heads were HP-treated for 10 min at different pressure levels (Frandsen et al, 2014), where a decrease in glucosinolate content was observed at 300 MPa, followed by an increase at 700 MPa.…”

Section: Accepted Manuscriptsupporting

confidence: 86%

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The impact of high pressure on glucosinolate profile and myrosinase activity in seedlings from Brussels sprouts

Wang

Barba

Frandsen

et al. 2016

Innovative Food Science & Emerging Technologies

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Section: Accepted Manuscriptsupporting

confidence: 86%

“…Extraction and isolation of myrosinase were performed as described by Bellostas, Petersen, Sørensen, & Sørensen (2008) with minor changes. Briefly, freeze-dried seedlings of Brussels sprouts (100 mg ± 0.1 mg) were ground into powder using mortar and extracted with Milli-Q-water (3 mL).…”

Section: Isolation Of Myrosinasementioning

confidence: 99%

The impact of high pressure on glucosinolate profile and myrosinase activity in seedlings from Brussels sprouts

Wang

Barba

Frandsen

et al. 2016

Innovative Food Science & Emerging Technologies

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“…To study the functional mechanism of these complexes their reassembly in vitro with recombinant or purified proteins or the extraction of intact complexes from plants (Bellostas et al 2008) need to be considered.…”

Section: Resultsmentioning

confidence: 99%

“…The M70 and M65 form, at least in vitro, complexes of high molecular masses with a group of proteins called myrosinase-binding proteins (MBPs) (Lenman et al 1990). Although the M75 member was thought to form homodimers but not complexes with MBPs (Lenman et al 1990;Eriksson et al 2002), a recent report indicates that even the M75 may form complexes (Bellostas et al 2008).…”

Section: Complexity Of Myrosinase Expression Patterns In Organs and Tmentioning

confidence: 99%

The ‘mustard oil bomb’: not so easy to assemble?! Localization, expression and distribution of the components of the myrosinase enzyme system

2008

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Glucosinolates are plant secondary metabolites that are hydrolysed by the action of myrosinases into various products (isothiocyanates, thiocyanates, epithionitriles, nitriles, oxazolidines). Massive hydrolysis of glucosinolates occurs only upon tissue damage but there is also evidence indicating metabolism of glucosinolates in intact plant tissues. It was originally believed that the glucosinolate-myrosinase system in intact plants was stable due to a spatial separation of the components. This has been coined as the 'mustard oil bomb' theory. Proteins that form complexes with myrosinases have been described: myrosinase-binding proteins (MBPs) and myrosinase-associated proteins (MyAPs/ESM). The roles of these proteins and their biological relevance are not yet completely known. Other proteins of the myrosinase enzyme system are the epithiospecifier protein (ESP) and the thiocyanateforming protein (TFP) that divert the glucosinolate hydrolysis from isothiocyanate production to nitrile/epithionitrile or thiocyanate production. Some glucosinolate hydrolysis products act as plant defence compounds against insects and pathogens or have beneficial health effects on humans. In this review, we survey and critically assess the available information concerning the localization, both at the tissular/cellular and subcellular level, of the different components of the myrosinase enzyme system. Data from the model plant Arabidopsis thaliana is compared to that from other glucosinolate-producing Brassicaceae in order to show common as well as divergent features of the 'mustard oil bomb' among these species.

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“…In the undamaged broccoli plant tissue, glucosinolates are separated from myrosinases (EC.3.2.1.147) by membranes, preventing the myrosinase isoenzymes from catalyzing the glucosinolate transformations to different bioactive products [Bellostas et al, 2008a;. Disruption of cell membranes, due to cell damage, processing or chewing, leads thus to contact between glucosinolates and the myrosinase isoenzymes, and in the presence of water this results in many different kinds of transformation products, depending on the type of glucosinolates present and the reaction conditions [Bellostas et al, 2007Bones & Rossiter, 2006].…”

Section: Unauthenticatedmentioning

confidence: 99%

Effects of Novel Processing Techniques on Glucosinolates and Membrane Associated Myrosinases in Broccoli

Frandsen¹,

Markedal²,

Martı́n-Belloso³

et al. 2014

Pol. J. Food Nutr. Sci.

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longed shelf-life by inactivation of micro-organisms and enzymes. HP/HT can at temperatures above 50 o C lead to pasteurization using pressure levels from 250-500 MPa [Wilson et al., 2008], while sterilization can be obtained with HP at temperatures higher than 70 o C . Focus is thus on the application of HP in combination with high temperatures (HT), which may lead to tissue compression, cell wall breakage and cell membrane disruption, depending on the HP and HT levels as well as the type of vegetable or fruit that are treated [Michel & Autio, 2002]. In the present study, where broccoli has been processed, the food quality is particularly connected with the enzyme-substrate (myrosinase--glucosinolate) system, where the different types of non-enzymatic and enzymatic catalyzed hydrolysis products give rise to effects from being health promoting to affecting the taste and odor [Hansen et al., 1995[Hansen et al., , 1997 Jeffery & Aray, 2009].PEF is considered a non-thermal treatment, which can lead to pasteurization at ambient temperatures using electrical field strengths of 20-30 kV/cm, and short pulses of 1-10 µs [Mosqueda-Melgar et al., 2008]. Application of short electric pulses may result in pore formation in cell membranes, and depending on the treatment intensity, time, electric field and pulse energy the membrane permeabilisation High pressure/high temperature (HP/HT) and pulsed electric field (PEF) treatment of food are among the novel processing techniques considered as alternatives to conventional thermal food processing. Introduction of new processing techniques with fast and gentle processing steps may reveal new possibilities for preservation of healthy bioactive compounds in processed food. However, effects on various food components due to autolysis and fast reactions prior to the applied HP/HT or PEF need to be considered as the total contribution of processing steps affects the obtained food quality. The present experiments were performed on broccoli (Brassica oleracea var. Italica) florets, purée and juice. Specific focus was given to effects of HP/HT and PEF processing on the content of glucosinolates and activities of myrosinase isoenzymes (EC.3.2.1.147) in the broccoli preparations. Certain conditions applied in HP/HT processing of broccoli florets were able to maintain a high level of intact glucosinolates. Treatment at 700 MPa and 20°C for 10 min was found to inactivate myrosinase activity, but also pressure treatments at 300 MPa and 20°C were able to maintain a high level of intact glucosinolates present in the untreated broccoli florets. PEF processing of broccoli purée and juice showed that the myrosinase activities resulted in nearly total glucosinolate transformations as result of autolysis during puréeing and juice making prior to the PEF processing. These data demonstrated that insight into potential effects on myrosinase activities from application of PEF processing implies specific focus on the sample steps preceding the PEF processing. Processing of broccoli and autolysis reaction...

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A fast and gentle method for the isolation of myrosinase complexes from Brassicaceous seeds

Cited by 26 publications

References 39 publications

The impact of high pressure on glucosinolate profile and myrosinase activity in seedlings from Brussels sprouts

The impact of high pressure on glucosinolate profile and myrosinase activity in seedlings from Brussels sprouts

The ‘mustard oil bomb’: not so easy to assemble?! Localization, expression and distribution of the components of the myrosinase enzyme system

Effects of Novel Processing Techniques on Glucosinolates and Membrane Associated Myrosinases in Broccoli

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