Principles of biochemistry, molecular biology, chemistry and chemical engineering are applied to: improve the nutritive value of soybean meal and reduce the environmental pollution from undigested phytic acid by use of phytase enzyme; enhance utilization of cereal, manure, and oilseed byproducts as adsorbent to remove toxic compounds found in waste water effluents; develop enzymatic processes to increase the reactivity of vegetable oils to produce industrial oils of added value; develop understanding of the mechanisms of decomposition of sucrose and other sugars during refining operations for food and fuel; and develop new and expanded uses of cottonseed products and by-products.

CRIS PROJECTS

The term "CRIS" stands for Current Research Information System and is simply the term we use when we talk about a research project. The following projects are funded through federal appropriations:

Develop Enhanced Phytases For Animal Feed And For Incorporation Into New plant Cultivars Requiring Less Phosphorus Fertilizer (CRIS 6435-13410-003-00D)

SCIENTISTS: Edward J. Mullaney and Abul H. Ullah

OBJECTIVES: First, develop phytases with significantly higher specific activity and broad substrate utilization by employing molecular biology techniques. Second, engineer higher heat stability in phytase and combine this with increased specific activity to produce a more cost effective enzyme for the animal feed industry. This will be followed by optimizing the enzymatic and nutritional properties of phytase for specific applications.

APPROACH: Analyze the sequence and molecular structural data on phytase molecules to achieve higher specific activity for phytic acid and then mutate the substrate specificity site and neighboring amino acid residues in A. niger NRRL 3135 phyA to effect these changes. These same techniques will also be employed to widen the variety of substrates that can be utilized. Increase heat tolerance in phytase will be effected by replacement of specific amino acid residues in the phyA molecule with amino acids occurring at higher frequency in stable proteins; this will result in a mutant phyA with increased heat tolerance. Once this goal is achieved, a combination of increased thermostability with the mutations conferring the higher specific activity will be undertaken.

Characterization And Improvement Of Sugar Industry Process Units Impacted By New Production Practices (CRIS 6435-41000-095-00D)

SCIENTISTS: Gillian Eggleston and Sarah E. Lingle

OBJECTIVES: To determine the physico-chemical properties of juice from sugarcane varieties subjected to new harvesting and storage practices. Develop markers/indicators of deteriorated and green (unburnt) sugarcane, based on physico-chemical properties, that can be used to predict processing problems. To determine the levels and effects of green trash that impact sugar processing, and improve harvest and processing methods to increase efficiency and reduce sugar losses.

APPROACH: Develop and deliver methods/tests to sugar factory personnel, which can be used to measure sugarcane quality indicator compounds that can predict future processing problems. Undertake studies to characterize the physico-chemical quality and processing ability of different components of green sugarcane trash. Undertake parallel field and factory studies to evaluate the effect of trash quality parameters on factory performance. Undertake collaborations with process managers and designers, to reduce the negative impact of green trash impurities on the efficiency of two specific industrial processes by improving process controls and designs and the use of processing aids.

New And Expanded Uses Of Oilseed Products And By-Products (CRIS 6435-41000-096-00D)

SCIENTISTS: Michael K. Dowd, Oliver Dailey Jr., and H. N. Cheng

OBJECTIVES: The objectives of this project are to (1)develop new industrial uses for cottonseed products and co-products as alternative fuel additives, biobased lubricants, and concrete-form release agents, (2) develop cost-effective methods for producing gossypol from cottonseed and cottonseed by-products, (3) develop new methods and processes for the conversion of oleic and linoleic acids, the most abundant fatty acids of cottonseed, to branched-chain, conjugated, and hydroxylated fatty acids, and (4) develop cost-effective methods for producing new value-added fatty acid-based products from cottonseed processing by developing new process methods for removing gosssypol and phospholipids from cottonseed oil miscella.

APPROACH: The research team will focus on developing value added uses and modified processes for cottonseed. This will be accomplished by developing cottonseed oil-based formulations that will be useful as fuel additives, lubricants, and concrete-form-releasing agents, by developing processes to remove gossypol from cottonseed miscella and processes for preparing gossypol from cottonseed and cottonseed products, by developing crystallization techniques for the preparation of chiral gossypol, and by developing new chemical approaches to convert linoleic or oleic acids into valuable conjugated, hydroxylated, and branched-chain fatty acids.

Enzymatic Processes For Increasing Industrial Utilization of Vegetable Oils (CRIS 6435-41000-087-00D)

SCIENTISTS: Jay Shockey, Heping Cao, and K. Thomas Klasson

OBJECTIVES: Identify enzymes involved in the biosynthesis of tung oil. Develop a microbial expression system for bioconversion of low-cost oils into tung-like drying oils. Optimize activity of tung enzymes in microbial bioconversion system.

APPROACH: Genes encoding the enzymes for tung oil biosynthesis will be identified by two complementary methods: 1. A homology-based approach in which tung genes are identified by their similarity to previously identified, oil-related genes characterized in other plant species. Gene-specific probes will be designed to screen a tung seed cDNA library by either PCR or traditional library screening. 2. A genomics approach in which all genes expressed during tung oil biosynthesis are first identified, then microarray or mass sequencing projects are used to identify oil related genes by bioinformatics methods. A microbial expression system tailored for the bioconversion of low-cost oils into tung-like drying oils will be generated by engineering common baker's yeast to secrete lipases into the growth media. The yeast cells will also be modified to co-express two or more tung enzymes to increase the conversion rate of exogenously acquired oils into tung-like drying oils. The activity of lipid-modifying enzymes will be optimized by studying the relationship between protein half-life and enzyme activity. Factors that increase or decrease enzyme activity will be identified and the information utilized to improve enzyme performance in yeast cells.

Agricultural By-Products as Adsorbents For Environmental Remediation (CRIS 6435-41000-088-00D)

SCIENTISTS: K. Thomas Klasson and Isabel Lima

OBJECTIVES: Research will be conducted on the conversion of agricultural residues, which include plant by-products and animal manures, 1) to develop activated carbons from poultry wastes, as adsorbents for inorganic contaminants such as metal ions and ammonia gas and 2) to develop strategies for the conversion of plant by-products to activated carbons and ion exchange resins that are specific for anionic pollutants.

APPROACH: Low value, high volume agricultural plant and animal waste will be converted to higher value products, such as activated carbon and ion exchange resins, intended for the commercial marketplace. The products will be used in environmental applications to improve air and water quality across the United States. Cost-effective processes will be developed for each product and a series of assessments will be made on the feasibility of establishing product markets for futurecommercialization. Two CRADAs have been completed, one with Grain Millers Inc, Eden Prairie, MN and another with Technology International Inc, LaPlace, LA to move forth the utilization of plant-based activated carbons. Additionally, agreements are in place with University of Maryland (for testing of adsorptive capability of animal waste- and pant-based activated carbon in sediment applications), SEKAB E-Technology (for creation of activated carbons from biorefinery by-product), and Alterna Energy (for developing custom carbons from plant waste. Collaboration is anticipated with additional companies to develop custom-made activated carbons from animal wastes and plants.