Implementation
Overview
Since the reform and opening up of China, animal husbandry has got outstanding achievements. The supply for products of husbandry animals has improved a lot, which changed from "overall undersupply" to "sufficient supply of aggregate." The market scale has gradually increased in recent years, and the demand for feed has also increased.
Figure 1: Changes in ruminant feed production since January 2018
Figure 2: From 2011 to 2017, the total production value of additives is increasing annually
However, animal feed mainly includes silage feed and cereal feed, silage contains a lot of cellulose, which is usually combined with hemicellulose, pectin, and lignin, making it difficult to digest and absorb. For cereal energy feed has high potential nutritional value as feed for poultry, the high content of soluble non-starch polysaccharides hinders the utilization and absorption of poultry. We aim to develop the product as feed additives to improve the utilization rate of silage and cereal through the multi-enzyme synergistic degradation strategy.
Target Customers
We divide the buyers by the quantities of ruminants on the farm. The farms with more than 3,000 cows are large-scale farms, and the farms with less than 3,000 cows are small-scale farms. For large-scale farms, there will be more cows with poor gastrointestinal systems, so it is worth buying the additive digestion aid. For instance, for those large-scale meat suppliers, their demand for feed additives is large- the reason is that the additive could help the cow grow better to get better meat. However, small-scale farms do not have much bargaining ability. Also, small-scale farms do not need to produce such a large scale.
To conclude, We choose three types of consumers, local retail, small feedlots, and local large feedlots.
Product
Our products as feed additives are able to help the animal gain more energy from the feed. Animals become more healthy, and with the increase in health value and better taste, the price of meat will rise.
We design the package of the products as shown below to make them more convenient to store. As seen in figure 3, our product for sale will be a pearl white plastic bag of white powder which weighs 1kg, what need to notice is we must keep our product in a dry condition and the temperature must be under 4 degree centigrade.
Figure 3. Proposed product package with our logo on it
Product Design Engineering
In order to improve the degradation rate of feed at the same time and solve the bottleneck problem of low efficiency and high cost of cellulase hydrolysis, we adopt the strategy of multi-enzyme collaborative degradation. Xylanase has the characteristics of hydrolyzing hemicellulose and can cooperate with cellulase to promote the biotransformation of lignocellulose. We further added the xylanase xynA gene of Bacillus subtilis into the cloning recombinant vector, it could be expressed with cellulase PKC-01 and β-xylosidase at the same time.
Figure 4. Multi-enzymatic synergistic degradation.
Safety
Safety is always the foremost priority we need to consider. Luckily there were no experimental accidents during our iGEM program. Due to the nature of our experiments, we used lots of enzymes and bacteria. These include E. coli Bl21, DNA T4 ligase, XynA, Xyl3A, CcxynA, restriction enzymes, DNA polymerase, PNC-5, and TOP10 competent cells. All of the enzymes we used are safe, active, and stable, and stored at appropriate temperatures. All lab equipment, such as Petri dishes, beakers, and graduated cylinders, were cleaned or sterilized before use.
Challenge
We are facing challenges from different aspects as below:

Challenges in development
1. How to reduce the degradation rate of the enzyme itself, (by optimizing the amino acid sequence)
2. How to improve the digestion efficiency of the enzyme (by studying the amino acid composition of the binding pocket, designing and improving the enzyme according to structural biology)
3. Investigate the ratio of enzyme to feed for maximum work efficiency
4. How to maximize enzyme production yield and minimize cost
5. How to design the PEGylation of enzymes so that animals can take them orally and lower the threshold for use
6. How to design and optimize enzyme-producing bacteria so as not to disturb the intestinal flora environment of animals
7. How to solve the decreased activity caused by lyophilization of enzymes
8. How to design and make the enzyme completely degraded in the body after the action is completed to reduce environmental pollution
9. How to design the ratio of 4 enzymes for optimal work efficiency
10. How to quantify the actual efficiency of enzymes in organisms and in feed
11. Genetically modified bacteria may affect the ecological environment
12. Although anti-degradation substances are added to the tablets compressed by enzymes, they may still be degraded in advance in the animal body or in the external environment, which cannot degrade the feed and allow the animals to better absorb nutrients.
13. If livestock eat too much, it may cause problems in their stomachs and intestines.

Legal challenges
The Chinese government has clear regulations on feed additives, as stated in the revised regulations on the administration of feed and feed additives issued by the State Council of the People's Republic of China in 2012.
6.2.1 Feed additives must be supervised and managed by the agricultural administrative department of the State Council.
6.2.2 The monitoring period of feed and new feed additives is 5 years.
6.2.3 There are workshops, equipment and storage facilities suitable for the production of feed and feed
additives,
and full-time technical personnel suitable for the production of feed and feed additives.
Above all, before our products are launched in the future, further testing and confirmation are required for the corresponding requirements.

Ethical challenges
For our product research, we resolutely do not use human tissue, cells, or animals for experiments. The use of enzymes can effectively avoid moral and ethical issues, and will not violate relevant regulations and laws, which will reduce the possibility of social controversy.
Our products use genetically modified engineering. Although many genetically modified products have appeared on the market, most people still have reservations. In order to overcome this difficulty, we will determine the safety of the products and constantly take efforts on promotion.

Marketing challenges
Since it is a genetically modified project and uses a lot of technological machines, the cost is high, which may lead to the high price of the products initially sold. However, we will gradually reduce the price of products through continuous improvement of technology, so that products become the choice of more people . The newly developed products may not be well-known and may not be trusted by users, which may cause the sales volume of our products to be inferior to those of other well-known brand companies. However, we will vigorously promote our products through the Internet and Offline promotion increases the visibility of products and companies.
Future plan
The focus of our future development is still on the commercialization of products, which we mainly discuss on two aspects, product development and marketing strategy.

In regard to the product development, per the challenges listed previously, we will consult more experts even cooperate with some relevant research group to overcome those challenges and optimize our products.

About the marketing strategy, we also drew a plan for this as shown below:
Price strategy
In the early stage of product sales, the products were first sold to larger domestic farmers at very low prices. It is best to achieve the purpose of buying in large quantities because the product is relatively cheap, and at the same time conduct small-scale sales in the market, which is also called the trial sale stage. After a long-term return visit after the trial sale, ask the farmers about the short-term and long-term consumption of cattle and sheep, whether there will be vomiting, abnormal excretion, or other conditions. Or whether it has the effect of promoting digestion and absorption, whether the effect is significant, etc. With the answer from the farmers, we will improve the product until the possibility of negative reactions of cattle and sheep is minimized as much as possible. And then sell it on a large scale. In the long-term product research and development, we will strive to reduce the price and create affordable and efficient products. This is to shape the brand reputation and company image, so that the company's products will become the choice of more people.
After-sales service
As a start-up, we will add after-sales service to the marketing plan to maintain the customers. When the buyer buys a small amount, we will give a gift bundled with the product, keep the contact information of the buyer, make regular return visits once or twice, listen to the follow-up evaluation of the product, and give gifts after the return visit. , call regularly to congratulate the purchaser on a happy holiday. Gifts are given at more important events. When the buyer purchases the sales volume stipulated by the company, we will reduce the unit price of the product, keep the contact information of the buyer, do regular playback three to four times, and also listen to the follow-up evaluation.
Promotion
For the ways of exposing our brand and our product, we divide it into online and offline ways.
Online, we could invite celebrities to promote our products. Meantime we will make use of various social media, like Bili Bili and Weibo to promote our product.
Offline, we could take our products to the agricultural fair, promote our products at the agricultural fair, and advertise our safety along with an expert who could answer questions from our potential buyers. In addition, we could also hold an event themed around our product to attract buyers.