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State University, AR, United States

Yactayo-Chang J.P.,Arkansas State University | Yoon S.,Arkansas State University | Yoon S.,University of California at Davis | Teoh K.T.,Arkansas State University | And 4 more authors.
New Negatives in Plant Science | Year: 2016

Background Expansin has been proposed to be an enhancer of cellulase activity in the deconstruction of biomass for sugars for industrial applications. However, the expansin protein is present in plant tissue only in minute quantities for promoting growth. Thus, producing adequate amounts of expansin for applications in industry will require a heterologous system that will over-express an expansin gene to produce large quantities of expansin protein. Development of a production system requires a facile, rapid assay. However, because no straightforward assay for expansin protein exists, we attempted to make milligram quantities of the protein in a fast or transient system for anti-expansin antibody preparation for use on Western blots or in ELISA assays. Results We tested the expression of the cucumber expansin gene in several heterologous systems including Escherichia coli and transient Nicotiana benthamiana leaves with limited success. We also had limited success in transiently expressing an alternative expansin gene from bamboo in N. benthamiana leaves. In order to determine if expansin over-expression is limited to a seed system, Arabidopsis thaliana seeds were tested. Although all positive and negative controls behaved as expected, none of these common systems expressed the expansin gene well. Conclusions Over-expression of cucumber expansin in three heterologous systems, E. coli, transient tobacco leaves, and Arabidopsis seeds was unsuccessful. The cause of this failure is not known. These results confirm the necessity of experimentally exploring several heterologous systems for protein production in order to find one with utility. © 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license. Source

Hood E.E.,Arkansas State University | Devaiah S.P.,Arkansas State University | Fake G.,Applied Biotechnology Institute, Inc. | Egelkrout E.,Applied Biotechnology Institute, Inc. | And 7 more authors.
Plant Biotechnology Journal | Year: 2012

Using plants as biofactories for industrial enzymes is a developing technology. The application of this technology to plant biomass conversion for biofuels and biobased products has potential for significantly lowering the cost of these products because of lower enzyme production costs. However, the concentration of the enzymes in plant tissue must be high to realize this goal. We describe the enhancement of the accumulation of cellulases in transgenic maize seed as a part of the process to lower the cost of these dominant enzymes for the bioconversion process. We have used breeding to move these genes into elite and high oil germplasm to enhance protein accumulation in grain. We have also explored processing of the grain to isolate the germ, which preferentially contains the enzymes, to further enhance recovery of enzyme on a dry weight basis of raw materials. The enzymes are active on microcrystalline cellulose to release glucose and cellobiose. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd. Source

A method for providing a service to assist in obtaining regulatory approval of a product includes using a computing device programmed to search at least one database of literature and programmed to identify data relative to determining substantial equivalence for the product to provide a first data set. The method further includes determining experimental data to collect for the product based in part on the first data set, collecting the experimental data for the product to provide a second data set, and documenting comparative data comprising comparisons between the first data set and the second data set data indicative of substantial equivalence for the product.

Garda M.,Arkansas State University | Devaiah S.P.,Arkansas State University | Devaiah S.P.,East Tennessee State University | Vicuna Requesens D.,Arkansas State University | And 7 more authors.
Transgenic Research | Year: 2015

Transgenic plants in the US and abroad generated using genetic engineering technology are regulated with respect to release into the environment and inclusion into diets of humans and animals. For crops incorporating pharmaceuticals or industrial enzymes regulations are even more stringent. Notifications are not allowed for movement and release, therefore a permit is required. However, growing under permit is cumbersome and more expensive than open, non- regulated growth. Thus, when the genetically engineered pharmaceutical or industrial crop is ready for scale-up, achieving non-regulated status is critical. Regulatory compliance in the US comprises petitioning the appropriate agencies for permission for environmental release and feeding trials. For release without yearly permits, a petition for allowing non-regulated status can be filed with the United States Department of Agriculture with consultations that include the Food and Drug Administration and possibly the Environmental Protection Agency, the latter if the plant includes an incorporated pesticide. The data package should ensure that the plants are substantially equivalent in every parameter except for the engineered trait. We undertook a preliminary study on transgenic maize field-grown hybrids that express one of two cellulase genes, an exo-cellulase or an endo-cellulase. We performed field observations of whole plants and numerous in vitro analyses of grain. Although some minor differences were observed when comparing genetically engineered hybrid plants to control wild type hybrids, no significant differences were seen. © 2014, Springer International Publishing Switzerland. Source

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