Legume Indian Bean (Phaseolus Vulgaris) Partially Purified Anti-Nutritional Protein Factor Exhibit Anti-Oxidant, Immunomodulatory and Anti-Cancer Properties

Abstract

Objective: This study was aimed to access one of the hidden ANF in Indian bean and assessment of its antioxidant, immunomodulatory, antiangiogenic, and anti-cancer properties. Methods: Indian beans were processed and evaluated for the presence of ANF through HA activity. Bean-ANF was partially purified by ammonium sulfate precipitation (ASP) and dialysis. Obtained Partially Purified bean-ANF (PPb-ANF) resolved on 12% SDS-PAGE, analyzed protease digestibility. The immunogenicity of PPb-ANF was characterized by producing polyclonal antibodies and checked for the presence of natural antibodies in healthy human sera. Pharmacological studies of PPb-ANF were assessed through anti-oxidant, immunogenic, anti-angiogenic and anti-cancer assays. Results: Partially purified Bean-ANF (PPb-ANF) showed significant specific HA activity up to 150 Units/mg compared to crude extract 9.3 of Units/mg. The PPb-ANF was identified to have more than 90% homogeneity in purity with an observed M W of 30-32 kDa. PPb-ANF showed significant pepsin digestive stability up to 2 h at 37 ᵒC and appears digestive resistant. Rabbit produced anti-PPb-ANF IgG polyclonal antibody on immunization and exhibited immunogenic properties. Natural human IgG and IgE antibodies were found in the non-atopic human sera against PPb-ANF, indicative of its immunogenicity. Pharmacological studies of PPb-ANF confirmed its potent antioxidant activity. At 400 µg of PPb-ANF showed the highest around 90% DPPH activity and at 150 µg of PPb-ANF showed 89% inhibitory lipid peroxidation activity. The PPb-ANF showed 70% (3 μg) and 40% (1.5 μg) inhibitory in blood vessel density compared to control and exhibited significant anti-angiogenic activity. The PPb-ANF showed a significant (*p<0.05.) reduction in solid tumor weight upon dose-2 treatment (50 mg/kg body weight) and also increased lifespan up to 70 d compared with the control tumor model. Conclusion: PPb-ANF was identified as digestive-resistant, hemagglutinin with a molecular weight of 30-32 kDa. The protein is immunogenic and has potential antioxidant, antiangiogenic and anti-cancer properties. Further purification and mechanistic-based pharmacological characterization might explore this protein as a potential anti-cancer molecule.

An Assessment of the Relative Antioxidant Activity of Bark Extracts of Acacia Catechu by in Vitro Free Radical Scavenging Analysis

“…Furthermore, Acacia catechu extracts have been shown to control the production of nitric oxide by peritoneal macrophages in a way that depends on the dose given. It has been discovered that the same stimulus elicits the secretion of IL-10, which is a crucial immunoregulator involved in inflammation, while simultaneously inhibiting the synthesis of TNF-α, a mediator of inflammation, that is released by monocytes and macrophages [17][18][19][21][22][23]. In a separate investigation, the researchers examined the efficacy of the 70% methanolic extract derived from the heartwood of Acacia catechu in scavenging several radicals, including nitric oxide, peroxynitrite, hydrogen peroxide, singlet oxygen, and hypochlorous acid [20,24,25].…”

A Comparative Study of Polyphenolic Content in Acacia Catechu Bark Extracts and Bibliographic Analysis With Reference to Guna (Madhya Pradesh), India

“…Although research has been thorough, the precise ways in which polyphenols work in the human body have not been definitively confirmed. However, there is compelling evidence indicating that certain targets including nitric oxide metabolism, carbohydrate digestion, and oxidative enzymes play a crucial role in providing health advantages [21,[23][24][25][26].…”

Evaluation of Comparative in Vitro Free Radical Scavenging Potential of Bark Extracts of Acacia Catechu

“…Although the use of herbal remedies is widely used throughout the world [3]. In almost all developing nations, including India, people mostly rely on local traditional medicine, which often includes herbalism, as their primary form of health care [14]. Notably, over 30% of pharmaceutical preparations continue to be derived either directly or indirectly from botanical sources [5,15].…”

“…The traditional medicinal practices of India have engrossed individuals by virtue of their enduring cultural significance and intergenerational transmission [14]. Moreover, the traditional system of herbal medicines for the prevention and treatment of various diseases is observed by over 80% of the people residing in the state of Madhya Pradesh, India [16][17][18].…”