Proteomic Analysis of Pollen and Blossom Honey from Rape Seed Brassica Napus L.

Abstract

A b s t r a c t In the study, honey from oilseed rape Brassica napus L., and both hand-collected (winter rape Visby and Cult) and bee-collected pollen of oilseed rape were analyzed for their proteome content, in order to see if any plant proteins were present to allow the proteo-typing of the oilseed rape honey. Proteins were fractionated by two-dimensional gel electrophoresis (2DE), stained by Coomassie blue and then analyzed by mass spectrometry. All identified proteins were divided into few groups due to their biological function. In 2DE gels with separated proteins from blossom honey, only bee (Apis mellifera) main proteins (Major royal jelly protein 1-5 and Glucosidase) were found. So we analyzed all proteins using gel-free based analysis with the SYNAPT G2 high definition mass spectrometry. We identified proteins that were present in both oilseed rape pollen and honey (Bna, Polygalacturonase, Non-specific lipid-transfer protein, GAPDH and others). We believe that these proteins are important for the nutritional value of plant pollen-enriched honey and further research is required on honey and honeybee pollen protein.Keywords: Brasica napus L., honey, honeybee pollen, proteomics, rape In recent years, there has been an increased demand for honeybee pollen as a healthy food supplement. The content of protein, fat, phosphorus, iron and vitamins (E, D, B 12 ) makes pollen's nutritive value comparable to dried beans or beef (Erlund, 2004). The unique properties of pollen were one of the reasons for using it as a pharmaceutical preparation. It was also important whether pollen was gathered by hand directly from the flower or by bees which is particularly rich in vitamins with a far higher concentration of pantothenic acid. The German Federal Board of Health has officially recognized the use of pollen as medicine; pollen has been helpful when administered in cases of chronic prostatitis (Cai et al., 2014; Wagenlehner et al., 2009). Some chronic diseases such as cancer, coronary, and neurological degeneration have been reported to be a consequence of oxidative damage (Babizhayev, Vishnyakova, & Yegorov, 2014; Jiang, Sun, & Chen, 2016). The therapeutic potential of honey is almost always associated with the antioxidant capacity against reactive oxygen species (Ferreira et al., 2009 Al-Meeri, & Al-Habori, 2002; Tonks et al., 2003;Brudzynski & Kim, 2011; Nasir et al., 2010; Erejuwa, Sulaiman, & Wahab, 2012). Oilseed rape (Brassica napus) is one of the most cultivated crop plants around the world. It is an important source of oil and medicinal components. The cultivation rate of oilseed rape around the globe has been increasing over the last ten years with about 31.5 Mha of lands used to cultivate in 2010 (Gulden, Warwick,& Thomas, 2008; Islam et al., 2013). Rape seed (Brassica napus L.) crops and honey are very good and commonly used bee food. Its pollen contains such amino acids as Threonine, Valine and Methionine and 23% to 24% crude protein, but the exact content is not exactly known. Honey differs in its com...

Floral nectar chitinase is a potential marker for monofloral honey botanical origin authentication: A case study from loquat (Eriobotrya japonica Lindl.)

“…This is consistent with previous findings that this protein is the dominant proteinaceous component in honey and prone to be degraded during the honey ripening and storage (Borutinskaite, et al, 2017;Di Girolamo et al, 2012). Failure of the 2-DE gel approach to detect floral proteins in honey is consistent with suggestions that these might be digested by honeybees, or degraded during honey ripening (Borutinskaite, et al, 2017;Di Girolamo et al, 2012). However, another possibility is that the concentration of floral nectar proteins in honey might be at an extremely low level, and/or obscured by high-abundant bee-originated proteins in honey when using routine proteomics approaches.…”

“…Among these, major royal jelly protein-1 from Apis mellifera was predominant, accounting for 19 spots from the 2-DE gel. This is consistent with previous findings that this protein is the dominant proteinaceous component in honey and prone to be degraded during the honey ripening and storage (Borutinskaite, et al, 2017;Di Girolamo et al, 2012). Failure of the 2-DE gel approach to detect floral proteins in honey is consistent with suggestions that these might be digested by honeybees, or degraded during honey ripening (Borutinskaite, et al, 2017;Di Girolamo et al, 2012).…”

“…On the gel, loquat honey proteins exhibited a series of zones estimated spanning the 5 to 100 kDa interval, the most abundant one was estimated to centre at 55 kDa. The profile was similar as other monofloral honey proteins, such as from chestnut, sunflower, orange, acacia and rapeseed (Borutinskaite, et al, 2017;Di Girolamo et al, 2012).…”

“…Therefore, the predominance of bee-origin proteins in honey may impede the detection of less abundant plant-origin proteins via regular gel-based approaches, and alternative methods may be needed. Using gel-free based analysis with high definition mass spectrometry, Brassica napus pollen proteins were identified in honey and deemed to be important for the nutritional value of plant pollen-enriched honey (Borutinskaite et al, 2017). However, very few studies have examined the existence of nectar proteins in honey, or explored their utility for honey botanical origin authentication.…”

Antibacterial potential of Swiss honeys and characterisation of their bee‐derived bioactive compounds

“…Unfortunately, the content of catalase and its enzymatic activity compared to GOX activity in different honey samples have not been determined yet. Surprisingly, identification of catalase in honey via proteomic approaches often failed …”