Team:SSTi-SZGD/Applied Design

SSTi-SZGD---Apply_Design

Overview

In the current Applied Design section, we described in details on how our project works. Started by a summary of core biological components that would be used in the formulation of products, then various aspects related to product design, including environmental impact, were considered. Later the target customers and marketing strategies were analyzed, and finally how our products could impact socially & economically, and areas for improvement were discussed.

Core Biological Component

Our wet lab results demonstrated that 1) LightOff system has relatively high induction efficiency, low leakage, and is feasible to be used to highly produce OPH or MHE 2) with the aids of E. coli Tat translocase, hydrolase could be secreted to the cell surface and work directly on pesticides; 3) expression of supernova gene may release ROS upon light irradiation to cause cell death. Pictures below summarize some key experimental results. For a full description of wet lab results, check Expression and Degradation pages. Once these elements were further verified and functionally optimized, we would put them together to create complete constructs to use either in industrial enzyme production, or directly as live biocatalysts.

Product design

Current Degradation methods

Pesticide residue degradation in soil is mainly achieved through natural degrading processes induced by sunlight, rainfall, heat or microorganisms. Solar radiation causes a cracking of chemical bonds of pesticide thus reduce toxicity. However pesticide stability and sunlight sensitivity varies. Rainfall physically removes soluble pesticide but can result to underground water contamination. High temperature removed pesticide by evaporation but it is limited by the volatile rate of pesticides. Microorganisms use pesticide or its secondary metabolites as carbon source, and it is the most efficient and cost-effective natural method. However specific pesticide degradation requires the availability of specific microorganism groups. Natural degradation approaches usually take a long time and the efficiency is very limited. On the other hand, domestic degradation of pesticide residues on fruits and vegetables can be achieved using chemical or physical methods. Chemical agents, although can thorough detoxify the surface of the food products, may cause secondary pollution, and their waste solutions are harmful to the environment. Physical methods, through the application of ultrasound, ionizing radiation and adsorption, require expensive equipment and expertise to operate, which is unrealistic to ordinary customers.

Our Proposed Products

Product name: Pesticide Residue Sweeper (R)

(suitable for organophosphate or carbendazim)

1) Enzyme products: Fast Acting Enzyme for pesticide residue removal

We plan to develop two forms of enzyme products: liquid form and powder form, which both contain active enzymes. Powder form is produced by spray-dried method, it retains the stable structure of enzyme and easy for transportation and storage. Our products targets customers in Great China region, which can be roughly divided as Northern China and Southern China with distinctly different local climate. Powder form is designed for users in Northern China region as in dry and cold condition, liquid is easily crystalized, layer separated or become frozen, while powder can be stored relatively well, and has better working efficiency in cold and dry environment. In contrast, liquid form enzyme product is designed for users in Southern China region as powder absorbs moist quickly and becomes solidified in hot and wet conditions. These two forms can both be diluted in water and sprayed onto the target object (automatically and manually).

2) Whole cell product: Super Whole cell for pesticide residue removal

In addition to enzyme products, we also designed ‘whole cell product’ that is a mixture of dried bacteria cells and dehydrated culture media. We also developed an automatic spraying system (refer to Hardware) that allows users to spray live bacteri a cells directly to the target object. The feasibility of ‘whole cell product’ relies on fact that a protein export component was engineered in the bacteria cells (refer to Degradation). This component allows the enzymes to be exported into periplasmic space, and further to be secreted in the cultural media.

The reason to manufacture whole cell product is to omit the needs having to manufacture enzyme products, which require stringent manufacturing procedure include ultrafiltration, concentration, low temperature drying, etc. A large amount of investment is needed to purchase expensive equipment, renting plant space and hiring labor force to embark on operation. On the contrary, the production of whole cell product requires bioreactors aside from standard laboratory equipment. The total investment cost is thus significantly reduced.

Product design considerations

1) In order to scale up for industrial production purpose, we need to cultivate engineered bacteria cultural in large size bioreactors and fermentor. A detailed production procedure and relevant financial cost analyses were illustrated in Business Plan.

2) Packaging size, active ingredient contents, usage area were designed by consulting to other similar products currently available in the market. For instance, a pack of 500 g enzyme product usually contains 5-10% active ingredient. This was designed to achieve both degradation efficiency of a designated area (i.e. one arce of land) and economical returns (refer to Business plan). We are still at very early stage of product formulation, further evaluation is necessary to increase return on investment rate.

3)For the use of whole cell product, as is used as live biocatalysts, it takes a considerable amount of time for cells to respond to darkness induction and express corresponded degrading enzymes. The current experimental results suggests 12-20 h is sufficient for this process to take place. We have included this in the consideration of Hardware design. Further improvements can be achieved by using a stronger promoter or introducing mutation.

Environment Impact

If our products can be put into use in the future, then we need to consider many biosafety issues, including human safety, potential harmfulness to the natural environment. Our enzyme product will be manufactured in a closely regulated industrial plant so that GMO rules and regulations should be complied to minimize contamination concern. In the contrary, our whole cell product works by spraying live bacteria cells to the target object, using an automatic spraying system.

This may result to live GMO exposure to natural environment, can be a health and environmental concern. Our currently solution is to add a suicide system that kills host cells when degradation is done, so that E. coli cells die as soon as completing their tasks. In addition, if we were to change host cell to gram-positive bacteria or other organisms that release hydrolase into the cultural medium, a 0.45um filter (refer to above) that traps bacteria cells will be added to the cultivation tank. In this case, only cultural media containing hydrolase will be released to the natural environment. These precaution approaches should reduce the risk of GMOs contamination to the greatest degree.

Market analysis

1) Target customers:

I, private garden/farm owners
Currently in China, private garden/farm start appearing in economical affluent areas, such as Pearl River Delta region and Long River Delta region. People with more buying powers are not satisfied with food products from mass scale production, as these products are considered to be pesticide residue contaminated and less nutrient. Given the circumstance, many of them developed their own vegetable/fruit garden on the building roof, backyard of the villa, or even purchase a piece of landfrom local farmers.

By growing their own produce (sometimes hiring farmers to do so), they are very conscious of soil/water quality, addition of pesticide and chemical fertilizer. In order to approach these potential customers, we plan to offer free soil/water testing service at the beginning to evaluate the circumstance of pesticide residual contamination on their land. Then we would recommend specific products for pesticide degradation according to the test results as well as offer professional advises on how to maintain pesticide residue free on the produce.

II, New Countryside
By referring to new countryside, we talk about those rural villages with local government officials who fully cooperated the concept of "Green mountains clean rivers are treasures" that is strongly promoted by current Chinese central government. Ordinary farmers are accustomed to use heavy loads of chemical fertilizer and various pesticides with fast and instant results. They are generally less interest in bio-degradable pesticides (due to slow response rate) and usually are little concerned with environmental issues. Heavily polluted soil and rivers are common scenes in many villages, and pesticide packages are scattered around the farm. However, with the introduction of concept of "New Countryside", officials of these villages took the initiatives to change the way farming has been done by using less polluted fertilizer, bio-degradable pesticides, or even not using any to some extent.

They are more interested in pesticide degradation products in order to improve soil quality and protect environment. The reason behind is that with improved land, water quality, and general environment, these villages get environmental subsidies from the central government that helps with local infrastructure development. We performed an evaluation test on their land, and discovered that considerable level of herbicide and traces of insecticides (organophosphate and carbendazim) can be detected. It is under this circumstance that the officials from these villages are interested in our products. Also in NanXi Village, it is the first time we came up with an idea to set up an automatic spraying system. With this system, we wished to help farmers degrading traces of pesticide directly on the soil and the crops right before the produces are picked for selling

III, small-medium size rural cooperative farmland
Rural cooperative farmland is a fast growing form of farming In modern Chinese agriculture. This type of farm cooperation has the advantage of attracting external funding source, therefore able to secure large scale land contract and purchase large scale modern machinery. Cooperative farm is usually operated like a business cooperation, and is more conscious of cost, return on investment and product quality. By visiting cooperative farmland and understood their needs, we added a few new features to our proposed automatic spraying system to fully incorporate the needs of small~medium size cooperative farmland.

2) Marketing strategy

A company website will be set up as well as settled in large, renowned B2B platforms (such as Alibaba, Amazon, etc) as branded shops for product promotion and direct sale channel (refer to Business Plan).. Ideally, customers can purchase our products through the following procedures

In the same time, we will try to make cooperation with other companies that manufacture inorganic fertilizer products in order to expand product portfolio, increase sales and generate profits.

3) SWOT Analyses of the products:

If we were to make viable products based on our techniques and promote them in current market, we would need to know the strengths and weakness of our products, as well as the opportunities and threats we are facing. Therefore a SWOT analysis comes in handy to provide a summary of our product features, and to help with developing strategies for product promotion.

Evaluation of Impact

Areas for Improvement

Our project primarily focuses on degradation of pesticide residues in soil, along with possible application on agricultural crops before harvest. There are a broad spectrum of pesticide residues exist in riverstream, underground water, fruits, vegetables, etc. Currently we only completed the degradation studies of two types of pesticides. In the future we are committed to develop a series of pesticide residue degradation products and related services based on different needs and requirements.

We endeavor to provide an all-round security from farm to dinner tables. However every degrading scenario requires different considerations. For instance, if we were to degrade pesticides on fruits and vegetables, GMOs on food safety would to be considered; if were to degrade pesticides in water, solubility of enzyme products would to be considered.