My presentations

RISE 2021 Presentation transcript

2021

• Rahman M, et al. Indexing and characterisation of a new class of protein from rapeseed Brassica rapa meal. Research in Science, Engineering and Environment Conference, 26-27 October 2021, Southern Cross University, NSW, Australia.

Transcript

0:00-0:26

Good morning, everyone.

0:27-0:55

This is Mahmudur Rahman. I am welcoming you to my presentation on possibilities of rapeseed as human food by indexing and characterisation of a new class of protein from rapeseed Brassica rapa meal. Rapeseed is commonly known Canola; rapeseed is also a close variety of mustard.

0:56-1:10

The world is suffering from scarcity of food. specially protein, that is one of the important components of food, costliest among the components, hard to grow and hard to obtain.

1:11-1:50

The topmost cause of human death in the globe is hunger. This is more than COVID 19 cases, cardiovascular disease, road accident and any other diseases. About 21,000 people die of hunger every day. In 10 second, one child dies due to hunger and malnutrition. Even in the US and Australia, the scenario is not completely good.

1:51-2:24

So everywhere in the world there are people are crying for the lack of food, including protein. It is not an easy task to supply lots of food for the world population of about 8 billion people. To back this up, to feed this huge amount of people, a continuous cultivation and production chain should be backed up.

2:25-2:50

So, plant-based proteins are highly thought to be a potential source of protein, they are also thought to be climate friendly, they require less amount of land area, less energy and they are proven as associated with less emission of greenhouse gases.

2:51-3:15

So, worldwide, there is a big shift of consumer behaviours from animal-based protein to plant-based protein. Research focus is pointed on the abundant and healthy protein sources. This abundant and healthy protein source in my study is Canola.

3:16-3:50

Canola is an oilseed crop that have huge amount of oil in its seeds. After extraction of oil, the residue is rich of protein, 40% of the total seed. So, the waste by-product could be a great source of protein for human. It is cultivated worldwide in huge amount. Australia is one of the countries cultivating Canola in broad acre. Australian Canola is mainly non genetically modified. It is cultivated in Australia as a rotational crop with wheat.

3:51-4:20

Science has proven that it is one of the healthiest cooking oils that is widely used through the globe. After extracting the oil in the industrial scale, the remaining residue that is a by-product, called as “Canola meal”, globally produced in huge amounts.

4:21-4:51

The most cultivated edible oilseed is soy. The oil extracted residue is utilised as human food as tofu, plant-based protein in burger, soy milk, soy sauce, in different ways. Whereas Canola meal is only utilised as livestock feed and soil fertilizer. There is no use as human food in past days.

4:52-5:24

Considering the fact that Canola proteins are cheap, they are abundant, have higher amino acid levels and higher protein efficiency ratio-that is the ration of protein we eat, and the amount of protein utilised to form the body-mass is called protein efficiency ratio. Canola meal has higher protein efficiency ratio, they contribute to build-up our body muscle.

5:25-5:33

These facts are summarised and published as a review at the beginning of this study.

5:34-6:15

In the study, we found that there is no well-developed method to analyse the proteins in Canola seed. We used an EMS mutant population for our analysis developed in University of Nottingham for our analysis. And we developed and optimised a method to analyse the proteins in a single seed. We used simple SDS-PAGE analysis and Werstern blotting and utilizing them-the optimized method is published in the journal-“Agronomy”.

6:16-6:59

Southern Cross Plant Science, Southern Cross University hosts the Brassica database. They have an in-house database of Brassica rapa seed proteins. We used that database to explore what proteins are present in the seed. We compared that data, which is technically called “BLAST”, we BLAST’ed the data with a well known plant-Arabidopsis thaliana, which genetic information is well studied for this plant. We compared them and some candidate genes were found in Brassica rapa.

7:00-7:18

To get confirmed about the protein genes, we bioinformatically did some comparison, which is called multiple sequence alignment. High degree of homology was found with the candidate protein genes with the known plant proteins

7:19-7:31

A phylogenetic relationship was studied to reveal the genetic architecture and genome evolution as well as to predict the functions of the protein genes.

7:32-7:44

Three-dimensional modelling was done for the candidate proteins and the outcome was published in the journal “Clinical and Experimental Allergy”.

7:45-8:11

Traditionally, mustard and rapeseed are used in the diseases where bacteria are involved. They are also used in the food preservation where also bacteria are involved. While we did the bioinformatic study, we find the candidate proteins have high sequence similarity and proteomic similarity with the antimicrobial peptides reported earlier.

8:12-8:26

So, we did some in-silico studies and we find amino acid sequence similarities. We also did molecular docking; it also shows that the proteins have high tendency to bind with the bacterial enzymes and inhibit their activities.

8:28-8:37

In in vitro analysis, we also found the proof antimicrobial activities on bacteria.

8:38-8:56

All of these outcomes justified the use of mustard and rapeseed in bacteria borne diseases. The study was published in Frontiers in Pharmacology.

8:57-9:19

As a next step, we analysed the EMS mutant seed samples, the single seed, there were about two thousand seed samples of different varieties. We studied them using mass spectrometric fingerprinting and we identified and characterized 323 major proteins abundant in the seed.

9:20-9:37

Earlier, it was known that Albumin and globulins are the major seed storage proteins. Seed storage proteins are the proteins that are stored in the seed, and they supply nutrition when the seed germinates. The seed storage proteins have high potential of use as human food.

9:38-9:48

We identified and characterized a new class of plant protein in rapeseed which is called vicilin and characterized the role of the identified protein in seed physiological processes.

9:49-10:18

To summarize these, the obtained information in this study could be used to determine the biophysics of protein structure, determine the three-dimensional structure, they give us the physiological function of the protein, we can get idea about the bioactivities of the proteins and finally, they give us the metabolic profile of the proteins. Utilizing the metabolic profile of the proteins, we can predict the use of the protein as human food.

10:19-10:31

Tailoring all the information, it is expected that we’ll get rapeseed based proteins in our plate as food very soon. The study was published in PLOS ONE.

10:32-11:08

The scenic beauty of the yellow mustard and rapeseed field, in near future, it is expected, not only fill our eyesi;: ght but also come and fill our plates. More than three thousand metric tons of healthy proteins will be added in our food chain and help to mitigate the cry of hungry people and bring back the smile to the hungry children and the population.

11:09-11:22

Thank you

Animation courtesy: Manitoba Canola Growers/What is Canola? Retrieved from https://canolagrowers.com/on-the-farm/what-is-canola/

Youtube link: https://www.youtube.com/watch?v=w1_H-TJP72U

Journal article published from this study:

i. Rahman M, et al., Shotgun proteomics of Brassica rapa seed proteins identifies vicilin as a major seed storage protein in the mature seed. PLoS ONE 2021, 16(7): e0253384. https://doi.org/10.1371/journal.pone.0253384.

ii. Rahman M, et al., A Single Seed Protein Extraction Protocol for Characterizing Brassica Seed Storage Proteins. Agronomy 2021, 11(1), 107. DOI:https://doi.org/10.3390/agronomy11010107.

iii. Rahman M, et al., Identification, characterization and epitope mapping of proteins encoded by putative allergenic napin genes from Brassica rapa. Clinical and Experimental Allergy, 50 (7): 848-68, 2020. https://doi.org/10.1111/cea.13612.

iv. Rahman M, et al., In silico, molecular docking and in vitro antimicrobial activity of the major rapeseed proteins-napin and cruciferin. Frontiers of Pharmacology, 11, Article 1340 2020. Published as a part of research Topic: Natural Antimicrobial Peptides: Hope for New Antibiotic Lead Molecules. https://doi.org/10.3389/fphar.2020.01340.

v. Rahman M, et al., Brassicaceae Mustards: Traditional and Agronomic Uses in Australia and New Zealand. Molecules 23 (1), 231. (Invited review for the Special Issue-Natural Products Research in Australia and New Zealand. https://doi.org/10.3390/molecules23010231.