Soybean is among the most abundant agricultural products in North Dakota (ND). Thus far, the major use of soybean is extracting soybean oil, which has become an important income of ND soybean farmers. After extracting the oil, the residual components, although containing food proteins, valuable polysaccharides, and chemicals of energy resources, are often utilized as animal foods. This is a tremendous waste given the huge market on food and energy industries and the high soybean yield in ND. The cause of such a waste is the difficulty in extracting these valuable components from the intense soybean cellulose network. By far, little success has been achieved, due to the lack of proper approaches to efficiently and rapidly degrade the cellulose with low-cost, minimal damage to the valuable portions, and no unwanted side-products. Here, we will develop a unique nanoscale materials that can overcome these barriers. The key is to immobilize all enzymes that can degrade the cellulose network on the surface of a recently developed crystal called Metal-Organic Framework (MOF). MOFs offer enhanced stability to the trapped enzymes so that the reaction can be carried out under elevated temperatures and acidic pHs to enhance the efficiency of enzymes. The use of enzymes generates no damage to any valuable components. The developed nanoscale crystals are easily recovered for reuse, which increases the cost-efficiency, and produce no pollution, contaminant, or metal-toxicity (because food-grade metals will be utilized). The proposed research will maximize the use of soybean crops for industry and improve the incomes of soybean growers.

1) Preparing nanocrystals with endo/exocellulase and xylanase encapsulated on MOF crystals;
2) Characterizing the structure, activity, stability, and reusability of the nanocomposites;
3) Optimizing experimental temperature and pH to rapidly degrade soybean cellulose network using the developed nanocrystals.

1) High-purity, stable, functional nanoscale crystals with endo/exocellulase and xylanase encapsulated on the surface;
2) Optimal reaction conditions to degrade cellulose/hemicellulose network of soybeans using the developed nanocrystals;
3) a new strategy to encapsulate multiple large-substrate enzymes for other biomass degradation and/or conversion reactions.

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Update:
Research conducted
The central goal of this project is to find out a way to better use one of the most important agriculture products at North Dakota (ND), soybean. We are focused on obtaining valuable sugar and food proteins from the soybean residuals after soybean oil production. To do so, we developed nanocrystals with three enzymes that degrade the key component of soybean oil residues, cellulose, hosted on them. We demonstrated the effectiveness of our approach on native cellulose materials and are heading toward large-scale production of our strategy.

Why the research is important to ND soybean farmers
Soybean is one of the most abundant products of ND. The major use of soybean is extracting oil. After the oil extraction, the residual soybean is mainly used as animal foods, which are generally considered low-valued. However, these residuals contain important sugar and food proteins hidden in the intense cellulose network of the residuals. If these valuable parts can be taken out from the cellulose network, the value of soybean can be maximized leading to increased income of local farmers/soybean industries.

Final findings of the research
We found it possible to keep all the key enzymes on several nanocrystals with different properties (size, shape, thermal stability, and magnetic properties) and proved that each enzyme is active on the nanocrystals. We were also successful in proving the long-term stability of the resultant nanocrystals. It is difficult to recycle and reuse the nanocrystals after a few reaction rounds though, which has been the bottleneck of this project. However, we fully believe that with a combination of several strategies, it is possible to overcome this problem.

Benefits/Recommendations to North Dakota soybean farmers and industry
We have demonstrated the possibility of degrading cellulose from soybean residuals. This suggests that for soybean industry, the residuals after soybean oil extraction should be collected, cleaned, and subjected for more options of applications. Perhaps a portion of the residuals can still be used as animal foods, yet the majority should be degraded by cellulose-degrading enzymes such shown in our work and subjected to sugar and food protein extraction. There is still a long way to go toward this goal. However, if succeed, this will lead to a significant increase in our local income as well as the attraction of food and energy companies to ND, resulting in an overcome increase the in local economy and employment.

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Developing Multi-Enzyme Metal-Organic Framework Nanocrystals for Rapid Soybean Biomass Conversion
Zhongyu Yang, Assistant Professor, Department of Chemistry and Biochemistry, North Dakota State University
Executive Summary

Research conducted
The central goal of this project is to find out a way to better use one of the most important agriculture products at North Dakota (ND), soybean. We are focused on obtaining valuable sugar and food proteins from the soybean residuals after soybean oil production. To do so, we developed nanocrystals with three enzymes that degrade the key component of soybean oil residues, cellulose, hosted on them. We demonstrated the effectiveness of our approach on native cellulose materials and are heading toward large-scale production of our strategy.
Why the research is important to ND soybean farmers
Soybean is one of the most abundant products of ND. The major use of soybean is extracting oil. After the oil extraction, the residual soybean is mainly used as animal foods, which are generally considered low-valued. However, these residuals contain important sugar and food proteins hidden in the intense cellulose network of the residuals. If these valuable parts can be taken out from the cellulose network, the value of soybean can be maximized leading to increased income of local farmers/soybean industries.

Final findings of the research
We found it possible to keep all the key enzymes on several nanocrystals with different properties (size, shape, thermal stability, and magnetic properties) and proved that each enzyme is active on the nanocrystals. We were also successful in proving the long-term stability of the resultant nanocrystals. It is difficult to recycle and reuse the nanocrystals after a few reaction rounds though, which has been the bottleneck of this project. However, we fully believe that with a combination of several strategies, it is possible to overcome this problem.

Benefits/Recommendations to North Dakota soybean farmers and industry
We have demonstrated the possibility of degrading cellulose from soybean residuals. This suggests that for soybean industry, the residuals after soybean oil extraction should be collected, cleaned, and subjected for more options of applications. Perhaps a portion of the residuals can still be used as animal foods, yet the majority should be degraded by cellulose-degrading enzymes such shown in our work and subjected to sugar and food protein extraction. There is still a long way to go toward this goal. However, if succeed, this will lead to a significant increase in our local income as well as the attraction of food and energy companies to ND, resulting in an overcome increase the in local economy and employment.

The key to improve the utilization of residuals after soybean oil extraction is to develop multienzyme/MOF nanocrystals that can stabilize multiple cellulose-degradation enzymes under elevated temperatures (to help loosen the cellulose network) and acidic pHs (to maximize the catalytic efficiency of involved enzymes). The accomplishment of the proposed research will benefit North Dakota soybean growers/farmers by bringing in food and energy industries and improving the overall local economy. In particular, the developed nanocrystals will maximize the use of the “waste” after soybean oil extraction to generate new food and energy resources. Considering the high soybean production and the large potential market of new energy resources and food industries, being able to maximize the use of soybeans will increase the income of ND soybean growers. It will also facilitate the cooperation of different industrial fields to take the best use of agricultural products, which will attract more companies to North Dakota and improve the local economy.