Sustainable L-asparaginase Production from Agricultural Waste: A Method for Utilizing Spent Grain

Abstract

The presentinvention discloses a sustainable method for producing L-asparaginase from agricultural waste, specifically spent grain, through microbial fermentation. This process not only enhances enzyme production but also addresses waste management issues in the agricultural sector.

Specification

Description:FIELD OF THE INVENTION:
The invention relates to biotechnology and enzyme production, specifically utilizing spent grain from the brewing industry as a substrate for L-asparaginase production. It addresses sustainable practices in enzyme manufacturing while promoting waste reduction, offering an eco-friendly solution for producing this essential enzyme used in cancer treatment.
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3. BACKGROUND OF THE INVENTION:
L-asparaginase is a crucial enzyme used in the treatment of leukemia and other malignancies. Traditional production methods often rely on expensive substrates and contribute to environmental degradation. Agricultural waste, particularly spent grain from brewing, presents an underutilized resource. This invention addresses the challenges of cost and sustainability in enzyme production by harnessing spent grain, which is typically discarded. By utilizing this waste product, the proposed method not only reduces environmental impact but also promotes a circular economy, making L-asparaginase production more economical and environmentally friendly. This innovation aligns with global efforts to enhance sustainability in biotechnological processes.________________________________________


4. OBJECTIVES OF THE INVENTION:
The primary objective of this invention is to provide ansustainable method for the production of L-asparaginase using various agricultural wastes. Specific objectives include:
1. Sustainable Production: To develop an eco-friendly method for producing L-asparaginase using spent grain, minimizing reliance on costly and non-renewable substrates.
2. Waste Valorization: To utilize agricultural waste effectively, reducing environmental impact and promoting a circular economy in the brewing industry.
3. Cost Efficiency: To lower production costs associated with L-asparaginase by employing low-cost substrates, making the enzyme more accessible for therapeutic applications.
4. Enhanced Enzyme Yield: To optimize fermentation conditions and microbial strains to maximize the yield of L-asparaginase from spent grain.
5. Contribution to Public Health: To provide a sustainable source of L-asparaginase for cancer treatment, thereby addressing the increasing demand for effective biopharmaceuticals.
5. SUMMARY OF THE INVENTION:
The invention provides a method for producing L-asparaginase using spent grain as a substrate. The process includes the following steps:
1. Collection of Spent Grain: Gathering spent grain from local sources.
2. Pre-treatment of Spent Grain: Physically and/or chemically treating spent grain to enhance the availability of nutrients.
3. Inoculation with Microorganisms: Introducing specific microbial strains capable of producing L-asparaginase.
4. Fermentation Process: Cultivating the microorganisms in a controlled environment to promote enzyme production.
5. Extraction and Purification: Isolating and purifying L-asparaginase from the fermentation broth.________________________________________

6. DETAILED DESCRIPTION OF THE INVENTION:
The method described in this invention is designed to produce L-asparaginase from agricultural waste. The detailed steps are as follows:
Step 1: Collection of Spent Grain: Spent grain is collected from breweries, ensuring it is fresh and free from contaminants.
Step 2: Pre-treatment of Spent Grain: Spent grain undergoes pre-treatment processes such as drying, grinding, or enzymatic treatment to enhance nutrient availability. This step may involve adjusting the pH or adding certain chemicals to facilitate microbial growth.
Step 3: Inoculation with Microorganisms: Microbial strains, such as E. coli or Bacillus subtilis, are inoculated into the pre-treated spent grain substrate. The choice of microorganism is based on its known ability to produce L-asparaginase efficiently.
Step 4: Fermentation Process: The inoculated mixture is incubated under controlled temperature and aeration conditions to promote the growth of the microorganisms and production of L-asparaginase. Optimal fermentation parameters (e.g., temperature, pH, time) are determined through preliminary studies.
Step 5: Extraction and Purification: After fermentation, the broth is centrifuged to separate biomass from the supernatant. L-asparaginase is extracted using techniques such as precipitation, dialysis, or chromatography.
This process results in better utilization of the agricultural wastes, efficient production of the enzyme, and lower amount of liquid wastes as compared to the traditional methods.
, Claims:1. I/We claim a method for producing L-asparaginase comprising:
• Utilizing spent grain as a substrate.
• Inoculating with a microbial strain capable of producing L-asparaginase.
• Optimizing fermentation conditions to maximize yield.
2. I/We claim the method of claim 1, wherein the microbial strain is selected from the group consisting of E. coli, Bacillus subtilis, and Aspergillus niger.
3. I/We claim a product of the method claimed in claims 1 or 2, wherein the L-asparaginase produced is suitable for use in cancer therapy.
4. I/We claim the method of claim 1, wherein the spent grain is pre-treated by methods selected from the group consisting of steaming, soaking, and enzymatic treatment to enhance enzyme yield.
5. I/We claim the method of claim 1, wherein the fermentation is carried out at a temperature range of 25°C to 37°C.
6. I/We claim the method of claim 1, wherein the pH is maintained in the range of 6.0 to 8.0 during fermentation.
7. I/We claim the method of claim 1, wherein the fermentation duration is optimized to be between 24 to 72 hours.
8. I/We claim the method of claim 1, wherein additional nutrients are added to the fermentation medium to support microbial growth and enzyme production, including nitrogen sources and carbon sources.

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