Purification and characterization of proteins in multifloral honey from kelulut bee (stingless bee)

Abstract

The kelulut bee (Meliponini) is a subfamily of stingless bees that produce honey. A total of 89 species out of a total of 500 species of kelulut bees are known to originate from the Indo-Australian region. Kelulut bees do not have quality standards so they still refer to the Codex and EU Directive which basically only applied for Apis honey. The Codex and EU Directive are formed by several psychochemical parameters, one of it is diastase activity. Diastase activity in kelulut honey is known not to meet existing standards or even undetectable. Therefore, this study aimed to explore proteins inside kelulut honey and investigate the possibility of using a specific protein as a biomarker to differentiate honey produced by kelulut bee from other honey. This research can also be considered as an initial step to optimize the exploration of protein in kelulut honey. This research is divided into two sections which are the preliminary research and the research expansion. From preliminary section, glucose dehydrogenase enzyme (GDH) was found to be present inside Tetragonula spp honey. A further examination of GDH enzyme was made in four kelulut bee honeys namely Tetragonula leaviceps, T. biroi, Heterotrigona itama, and Geniotrigona thoracica. The preliminary research has five stages that are exactly the as expansion research section except it didn't include GDH activity measurement. The research includes seven main stages. First honeys were dialyzed to remove the sugar content followed by centrifugation. The samples were then purified using liquid chromatography with anion exchanger column. The molecular weight of proteins was analysed by SDS-PAGE method. The GDH activity was measured using spectrophotometer followed by qualitative analysis using LC-MS/MS. The peptide sequences resulted from LC-MS/MS were then matched with Uniprot to identify the unknow protein. The results showed that only T. biroi and T. laeviceps had GDH enzyme activity of 0,1891 U/mL and 0,1652–1,579 U/mL, respectively. Bands from both species were also qualitatively identified as GDH. With these results, it can be concluded that the GDH enzyme cannot be used as a biomarker to distinguish the kelulut honey.

Current Trends and Technological Advancements in the Study of Honey Bee-Derived Peptides with an Emphasis on State-of-the-Art Approaches: A Review

“…Table 3. Different factors that impact the separation and purification of honey bee peptides [35,37,47,62,[65][66][67][68][69][70][71][72][73][74][75][76][77][78].…”

Determination of Antioxidant Activity, Total Phenolic and Flavonoid Contents of Malaysian Stingless Bee Propolis Extracts

“…Stingless bees are known locally as "lebah kelulut" are members of the Apidae family and the Meliponini tribe with approximately 500 identified species from 32 genera across the globe (Sahlan et al, 2019). Stingless bees lack a functional sting and are of little harm to bee-keepers as opposed to honeybees (Asem et al, 2019).…”

Effect of Thermal Treatment on Kelulut Honey Towards the Physicochemical, Antioxidant and Antimicrobial Properties

“…In Malaysia, stingless bee honey is commonly referred to as Kelulut honey. There are many different species of stingless bee honey, namely Heterotrigona erythrogastra, Tetragonula biroi and Heterotrigona itama (Sahlan et al, 2019). Kelulut honey has higher antioxidant and antimicrobial properties compared to Tualang honey (Sulaiman & Sarbon, 2020).…”