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Global Research journal of Natural Science &  
Technology (GRJNST)  
Volume: 04 - Issue 2 (2026), 2071  
ISSN P: 2790-7643 ISSN E: 2790-7651  
Biochemical characterization and profiling of major seed storage proteins in  
selected medicinal plant found in Sindh Pakistan  
Received: 25 March 2026 Accepted: 10 April 2026. Published: 27 April 2026  
S.K. Khanzada  
Institute of Plant Sciences, University of Sindh  
AK Khanzada  
Institute of Plant Sciences, University of Sindh  
Beenish khanzada  
Institute of Biochemistry University of Sindh  
Shaheen Memon  
Institute of Plant Sciences, University of Sindh  
B. Sadiq  
Institute of Plant Sciences, University of Sindh  
Imran Ali Bhurt  
Institute of Plant Sciences, University of Sindh  
Abstract: In this study investigated the composition and biochemical properties of  
seed storage proteins in important medicinal plants, including the seeds of  
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Syzygium cumini L., Lawsonia inermis L., Calotrpis procera. R Bro, Ricinus  
communis L., Linum usitatissimum L., Brassica juncea L Czern., Withania  
coagulans Dun, and Cucumis melo var. agrestis Naudin. Protein extraction from  
seed flour was carried out using sequential steps of extraction: delipidation (removal  
of oil), water (albumin), 5.0 M NaCl (globulin), 70% ethanol (prolamin), and  
150.2M sodium phosphate buffer, pH 8.0 (glutelin). Quantification was performed  
using the dye binding method of Bradford which showed substantial differences in  
their concentrations and total yields. Comparative analysis of seed storage proteins  
showed different dominance patterns among the plants studied. The highest levels  
of albumin were found in Brassica juncea L Czern. (50.40%), which suggests a  
strong water-soluble protein fraction, the highest globulin content was found in  
Ricinus communis (67.11%), suggesting a high salt-soluble protein fraction, the  
highest prolamin content was found in Withania coagulans (55.93%), suggesting a  
strong alcohol-soluble protein fraction, and the highest glutelin was found in  
Syzygium cumini (15.12%).  
Keywords: Seed storage proteins (SSPs) Extraction, Protein estimation, Medicinal  
plants  
INTRODUCTION  
During seed germination and the early stages of seedling growth, seed storage proteins serve  
as important stores of nitrogen and amino acids. These proteins are classified based on their  
solubility and structural properties into albumins, globulins, prolamins, and glutelins. In  
addition to their physiological role in plants, seed storage proteins have become more  
interesting to study due to their nutritional value and bioactive activities, such as  
antibacterial, antioxidant, and medicinal properties (Yang et al., 2023; Zeng et al., 2024),  
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Recent studies have also emphasized the importance of medicinal plant seeds as a source of  
bioactive peptides and functional proteins with pharmacological applications (Latif &  
Nawaz, 2025). The seeds of Syzygium cumini L. rich in albumins and globulins and are  
well known for their antioxidant and antidiabetic activities, which are attributed to a variety  
of bioactive chemicals and protein fractions (Latif & Nawaz, 2025). The presence of high  
levels of storage proteins, especially albumins, and its hepatoprotective and antibacterial  
properties make Lawsonia inermis L. an important plant in traditional medicine (Kumar et  
al., 2024). A number of protein components, including defense-related and enzymatic  
proteins, have anti-inflammatory and antibacterial properties in the seeds of Calotropis  
procera R. Br. (Tiwari et al., 2022). Ricinus communis L. seeds contain both toxic proteins  
such as ricin and storage proteins such as globulins, but the seeds can be used medicinally if  
properly prepared (Yang et al., 2023). The seeds of Linum usitatissimum L. contain  
globulins and albumins and are used medicinally for their cardioprotective, antioxidant, and  
anticancer activities, in part due to the composition of their proteins and related metabolites  
(Zeng et al., 2024). Due to their intricate biochemical composition, Brassica juncea L  
Czern., seeds contain antibacterial and chemoprotective compounds and high levels of  
globulins (Yang et al., 2023). Research on storage protein albumin, globulin, prolamin,  
and glutelin. of Apiaceae Family medicinal plant, seeds are valuable, nutrient sources of  
seed proteins ,these seeds are utilized in home remedies to manage gastrointestinal issues  
such as indigestion, flatulence, and colic pain, while also acting as flavoring, aromatic, and  
preservative agents in food supplies (S.K. Khanzada et al. 2021), Bioactive proteins and  
secondary metabolites like withanolides are reported to be responsible for the antidiabetic  
and immunomodulatory properties of Withania coagulans Dun. seeds (Latif et al , 2025).  
Storage proteins are also important from a nutritional standpoint, and seeds of Cucumis  
melo var. agrestis Naudin. provide antioxidant and anti-inflammatory properties for  
traditional therapeutic use (Kumar et al., 2024). They have been investigated as a source of  
food and pharmaceuticals due to their biochemical composition, nutritional importance, and  
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medicinal properties, including medicinal plant seed storage proteins (M. Kumar et al.,  
2022). In human metabolism, seed storage proteins exhibit a wide range of activities, and  
albumins and globulins are abundant in angiospermic plants, including monocots and dicots,  
such as cereals, ferns, and palms (Templeman et al., 1987). Glutelins, on the other hand, are  
resistant to pepsin digestion and thus less digestible by humans. The biochemical  
characterization of (SSPs) in medicinal plants is important for understanding their  
nutritional value and therapeutic applications in the food and pharmaceutical industries.  
SSPs are simple biomolecules that are necessary for the growth, development, and  
reproduction of plants. (López-Guzmán et al., 2025). In this study investigated the  
biochemical properties and protein profiles of seed storage proteins in eight medicinally  
important plant species of various families.  
MATERIAL AND METHODS  
Material: Seeds of eight medicinally important plants belonging to the different plant  
families: Syzygium cumini L., (Myrtaceae), Lawsonia inermis L., (Lythraceae), Calotrpis  
procera. R Bro (Asclepiadaceae), Ricinus communis L., (Euphorbiaceae), Linum  
usitatissimum L., (Linaceae) Brassica juncea L Czern., (Brassicaceae), Withania coagulans  
Dun (Solanaceae), and Cucumis melo var. agrestis Naudin. (Cucurbitaceae). were collected  
from three different localities, Nawabshah, Hyderabad, Jamshoro University of Sindh  
Jamshoro for conducted to investigate the composition, biochemical properties of seed  
storage proteins in important medicinal plants.  
Seed storage protein extraction: The seed were ground with mortar pistil, the flour was  
delipidated by solvent extraction by stirring with pure hexane (1:10 w/v) for 30 min, and  
this procedure repeated three times at room temperature. Using the microextraction method  
(Sammour et al., 1999), the sample was air dried and kept under hood up to twenty-four  
hours in alcohol, water, high salt, and in alkaline buffer conditions. Finally, the seed was  
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extracted in triplicate in Eppendorf tubes with 500 micrograms (i) deionized water (ii)  
145.0M NaCl (iii) 70% ethanol and (iv) 0.2 M, NaPObuffer, pH8.0, to extract the four  
major seed storage proteins (prolamin, albumin, glutelin, and globulin), each at 500  
micrograms, for 20 min at 350 rpm/25C using Thermomixer comfort (Eppendorf,  
Germany). This extraction was performed at 14000 rpm for 15 min at 4ºC (Biofuge Primo  
R Heraeus, Japan) and repeated twice to remove residual protein from each fraction. The  
supernatants of each fraction were pooled and stored at -20C until protein profiling. The  
total protein from the entire extraction was measured using the modified color binding assay  
(Bradford, 1976) with bovine serum albumin as a standard. Protein quantitation was  
performed in triplicate using a microplate reader (Sunrise Tecan, Austria).  
Results and Discussion  
The seed storage protein content of eight medicinal plant species from Sindh, Pakistan is  
presented in Table 1. The results show noticeable variation among the species in the four  
main protein fractions, namely albumin, globulin, prolamin, and glutelin. In this study  
investigated the biochemical properties and protein profiles of seed storage proteins in eight  
medicinally important plant species of various families. S.cumini L., L. inermis L., C.  
procera. R Bro, R. communis L., L. usitatissimum L., B. juncea L Czern., W. coagulans Dun,  
and C.melo var. agrestis Naudin..These differences indicate that each species is composed of  
a different protein set, which could make it more or less nutritious and play different  
biological roles. Albumin showed a wide range of values (22.88 to 50.40), with B. juncea L  
Czern having the highest content (50.40), followed by C. procera. R Bro, (44.40), L.  
inermis L., (41.86), L. usitatissimum L., (40.14), and W. coagulans Dun (31.40), and the  
lowest in R. communis L (22.88). This high albumin proportion in B. juncea L Czern  
indicates it may be a good source of water-soluble proteins with high digestibility. Globulin  
values varied considerably, from 1.70% in L. inermis L. to 67.11% in R. communis L.,with  
C. procera. R Bro, (36.02%) and L. usitatissimum L., (34.82%) also exhibiting significant  
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globulin levels. In a previous study, 43.25% protein recovery and 17.2% extraction  
efficiency was reported for L. usitatissimum L., protein extracted at pH 12 (Sharma et al.,  
2022). However, the protein fractions differed and the major seed storage proteins were  
albumin (40.14%), globulin (34.82%), prolamin (22.38%), and glutelin (11.57%). L.  
inermis L., (1.70%) and W. coagulans Dun (4.29%) contained the lowest amounts. These  
differences represent species-specific storage strategies and can impact seed nutritional  
quality and industrial applications. Prolamin was the major protein fraction in many species,  
W. coagulans Dun (55.00%), L. inermis L., (53.32%), and S.cumini L (48.00%) followed  
by Cucumis melo var. agrestis (39.80%), B. juncea L Czern (26.56%), R. communis L  
(5.16%), and C. procera. R Bro, (7.75%). The high prolamin content in certain species  
suggests potential functional roles in seed physiology and possible applications in food and  
pharmaceutical industries, although prolamins are generally less nutritionally balanced due to  
low lysine content. Glutelin content ranged from 3.10% to 15.12%, with S.cumini L  
(15.12%) and C.melo var. agrestis Naudin (14.42%) showing the highest levels. Moderate  
amounts were observed in C. procera. R Bro, (11.81%) and L. usitatissimum L., (11.57%),  
whereas L. inermis L., (3.10%) and B. juncea L Czern (3.43%). The distribution of these  
proteins indicates that albumin and prolamin are the major proteins in most species, and  
that globulin is the major protein in R. communis L., (SSPs) storage protein composition is  
highlighted by the observed variation in protein fractions, which may be impacted by genetic  
determinants, ecological adaptability, and seed functional needs.The extent to which the  
proportions of these proteins differ between species may have significant nutritional quality  
and industrial applications.  
Table 1. Comparison of the concentrations and total percent yield of the major seed storage  
proteins from Syzygium cumini L., Lawsonia inermis L. Calotrpis procera.R Bro Ricinus  
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communisL., Linum usitatissimum L., Brassica juncea L Czern., Withania coagulans Dun,  
and Cucumis melo var. agrestis Naudin. seeds found in Sindh, Pakistan.  
S.NO. Plant Name  
FamilyName  
Globulin *  
Prolamin *  
# mg/g  
%yield  
Glutelin *  
#
Albumin  
#
mg/g %yield  
mg/g %yield  
mg/g %yield  
1.  
2.  
3.  
4.  
5.  
6.  
7.  
Syzygium cumini Myrtaceae  
0.83 30.53 0.19  
0.61 41.86 0.02  
0.13 44.40 0.10  
1.86 22.88 5.47  
1.34 40.14 1.17  
0.76 50.40 0.29  
0.08 31.44 0.01  
7.00  
1.30  
0.78  
48.00 0.40  
15.12  
L.  
Lawsonia inermis Lythraceaea  
1.70  
53.32 0.04  
3.10  
11.81  
4.81  
11.57  
3.43  
9.32  
L.  
Calotrpis  
Asclepiadaceace  
36.02 0.02  
67.11 0.42  
34.82 0.75  
19.60 0.40  
7.75  
5.16  
0.03  
0.39  
procera.R Bro  
Ricinus communis Euphorbiaceae  
L.  
Linum  
Linaceae  
22.38 0.38  
26.56 0.05  
55.00 0.02  
usitatissimum L.  
Brassica juncea L Brassicaceae  
Czern.  
Withania  
Solanaceae  
4.29  
0.15  
coagulance Dun.  
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8.  
Cucumis melo  
L.var. agrestis  
Cucurbitaceae  
0.03 30.77 0.01  
15.00 0.05  
39.80 0.01  
14.43  
Naudin.  
* Concentrations in mg/g of seed flour. Values are mean of three independent extractions.  
#
% yield of a particular protein in total protein contents of seed flour.  
Table 2: Seed storage protein (SSPs) composition, functional importance, and medicinal  
significance of selected medicinal plants found in Sindh Pakistan  
Medicinal plants With  
Importance of protein  
Medicinal significance  
family name  
Provide essential amino acids and  
support antioxidant defense  
mechanisms in human metabolism  
Syzygium cumini  
(Myrtaceae)  
Antidiabetic, anti-inflammatory,  
antioxidant  
Antimicrobial, antifungal,  
hepatoprotective  
Easily digestible proteins (albumins)  
enhance nutrient absorption and  
immune support  
Lawsonia inermis  
(Lythraceae)  
Defense-related proteins may exhibit  
bioactive and therapeutic properties  
in small doses  
Calotropis procera  
(Asclepiadaceae)  
Anti-inflammatory, analgesic  
Storage proteins include potent  
bioactive molecules; require  
detoxification for safe use  
Ricinus communis  
(Euphorbiaceae)  
Laxative, antimicrobial, anti-  
inflammatory  
Linum usitatissimum  
High-quality proteins contributes to Cardioprotective, antioxidant,  
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(Linaceae)  
cardiovascular health and metabolic  
regulation  
anticancer  
Rich in functional proteins (e.g.,  
cruciferin) supporting detoxification  
and cellular protection  
Brassica juncea  
(Brassicaceae)  
Antimicrobial, anti-inflammatory  
chemoprotective  
Bioactive proteins and peptides aid  
in immune modulation and glucose  
regulation  
Withania coagulans  
(Solanaceae)  
Antidiabetic, hepatoprotective,  
immunomodulatory  
Antioxidant, anti-inflammatory,  
diuretic  
Nutritionally important proteins  
contribute to hydration balance and  
metabolic support  
Cucumis melo var.  
agrestis (Cucurbitaceae)  
Cucumis melo L.var.  
agrestis  
Antioxidant, anti-inflammatory,  
diuretic  
Nutritionally important proteins  
contribute to hydration balance and  
metabolic support  
Naudin.  
Table 2 showed these medicinal plants are very valuable for human nutrition and  
therapeutic applications, seeds contain four types of protein (SSPs) that provide a balance  
of rapidly digestible proteins (albumins) and slow release, long-lasting proteins (globulins,  
prolamins, glutelins) to meet immediate metabolic needs, long-term amino acid supply, and  
delivery of bioactive compounds, which are directly linked to the observed antioxidant,  
antidiabetic, immunomodulatory, and cardioprotective activities of these plants. L.  
usitatissimum L., and L. inermis L. seeds are rich sources of albumin and globulin and,  
therefore, are especially important for human nutrition and therapeutic use.  
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Fig. 1. Graphical representation of the four major seed storage protein (SSPs)(Albumin,  
Globulin, Prolamin and Glutelin.) concentrations (mg/g) measured from Syzygium  
cumini L., Lawsonia inermis L. Calotrpis procera.R Bro Ricinus communisL., Linum  
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usitatissimum L., Brassica juncea L Czern., Withania coagulans Dun, and Cucumis melo  
var. agrestis Naudin. seeds found in Sindh, Pakistan.  
Conclusion  
The findings of the present study reveal that the composition of seed storage proteins differs  
markedly among the selected medicinal plant of Sindh, Pakistan. Albumin was found to be  
the dominant protein fraction in B. juncea L Czern, suggesting its key role as a primary  
water-soluble storage protein in this species. Contrastingly, R. communis L is highest  
amount of globulin, suggesting an abundance of salt-soluble proteins. W. coagulans Dun.  
had a unique protein profile, with prolamin being the dominant fraction, suggesting an  
adaptive mechanism of protein storage. S.cumini L had relatively higher levels of glutelin  
compared to the other species, but glutelin was always the least abundant fraction. This  
indicates that seed storage protein profiles are species-specific and influenced by genetic and  
physiological factors, and albumin, globulin, prolamin, and glutelin fractions vary in relation  
to seed structure, nutritional potential, and metabolic functions.  
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