TITLE:
Enhancement of the Essential Amino Acid Composition of Food Crop Proteins through Biotechnology
AUTHORS:
Godson O. Osuji, Eustace Duffus, Paul Johnson, Sela Woldesenbet, Aruna Weerasooriya, Peter A. Y. Ampim, Laura Carson, Yoonsung Jung, Sanique South, Edna Idan, Dwiesha Johnson, Diadrian Clarke, Billy Lawton, Alfred Parks, Ali Fares, Alton Johnson
KEYWORDS:
Plant Proteins, Citric Acid Cycle, Glycolysis, Glyoxylate Cycle, HPLC, Glutamate Dehydrogenase-Synthesized RNA, mRNA Co-Silencing, Mineral Nutrients, Permutation of Biochemical Pathways
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.6 No.19,
December
10,
2015
ABSTRACT: Lack of essential amino acids (EAA) in the diet of at-risk populations could beget a state of food
insecurity. Plant proteins are deficient in some essential amino acids. Animals obtain EAA from
plant sources. Simple biotechnologies are being developed for improving the EAA composition of
crop proteins. The aim was to integrate-discriminate glycolysis and citric-glyoxylic acid cycles to
optimize biosynthesis of EAA in food crops. Permutation of diverse metabolic pathways at the
mRNA level by glutamate dehydrogenase (GDH)-synthesized RNA is a common biotechnology for
doubling the nutritious compositions of plants. Peanuts were planted in plots and treated with
mineral salts mixed according to stoichiometric ratios. Protein-bounded and free amino acids of
mature peanut seeds were determined by HPLC. GDH-synthesized RNA probes homologous to the
mRNAs encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate mutase
(PGlycM), phosphoenolpyruvate carboxylase (PEPCase), enolase, malate dehydrogenase (MDH),
isocitrate lyase (ICL), and malate synthase (MS) were prepared from peanut seeds using restriction
fragment double differential display PCR method. Northern assays of peanut total RNA showed
that the mRNAs encoding PGlycM, PEPCase, MDH, and MS shared extensive sequence homologies
that produced a dense network of cross-talks, resulting to co-differential silencing of the mRNAs
thereby permuting glycolysis, citric-glyoxylic acid cycles. There were 42 permutations in the NPPKtreated,
105 in control, 420 in KN-, and NPKS-treated peanuts. Because of permutations involving
the mRNAs encoding ICL and MS, wherever the abundances of these mRNAs were high (control,
and NPPK-treated peanuts) the concentrations of the α-ketoglutarate group of total glutamate,
glutamine, arginine, proline, and histidine were minimized (28.0 mg/g). The integration of glycolysis, citric and glyoxylic acid cycles increased the
quality and doubled the concentrations of the protein-bounded EAA composition of NPPK-treated
(33.37 mg/g) compared with the control peanut (15.66 mg/g). The commanding biotechnology
was the stoichiometric mineral salts-based induction of GDH to synthesize the RNAs that integrated
glycolysis, citric-glyoxylic acid cycles to one functional unit.