Corn Cell Wall Research Articles

SIFAT KIMIA TEPUNG JAGUNG LOKAL PUTIH TIMOR TERMODIFIKASI MELALUI FERMENTASI BAKTERI Lactobacillus casei

The characteristics of flour much determine the quality of food products. Maize starch has several limitations to be developed into food products. Hence, to expand its use in food production, the starch needs to be modified. Modification of the starch can be done through the fermentation process. This modification is expected to change its physico-chemical aspects and functional properties. Some lactic acid bacteria (including L. bulgaricus and L.casei) are capable of degrading the corn cell wall, resulting in the starch granules to come out of the cell which eases the grinding process. L. bulgaricus and L. casei also can degrade starch contained in the growth medium into simple sugars, degrade proteins, and peptides into amino acids (Ganzle et al., 2008). The modified maize starch is also expected to be a proper substitution of regular flour in food products. The purpose of the research is to examine the quality of the modified starch through fermentation. The results showed that the fermentation time and concentration of bacteria dramatically affect the physical results where fermented corn was whiter and slightly acidic and chemically decreased the pH of corn flour to 4.5 and was relatively stable at 48 hours fermentation with a concentration of 4% and 6 %. The bestmodified chemical properties of local white corn are 36 hours fermentation with a concentration of 2% lactobacillus casei with a total protein value of 8.7%.

Cell Wall Characteristics of a Maize Mutant Selected for Decreased Ferulates

The cross-linked nature of plant cell walls provides structural integrity for continued growth and development, but limits degradation and utilization by ruminants. In grasses a major cross-linking component is ferulic acid that is incorporated into cell walls as an ester linked residue on arabinoxylans. Ferulates can become coupled to each other and to lignin forming a highly cross-linked matrix of carbohydrates and lignin. Seedling ferulate ester mutants (sfe) were produced in maize using the transposon system and evaluated in feeding trials. The work described here was undertaken to characterize changes in the ferulate cross-linked nature as well as other components of the corn cell wall matrix in leaf, sheath and stem tissues. Total ferulates decreased modestly due to the mutation and were more apparent in leaf tissue (16% - 18%) compared to sheath (+5 to -6% change) and stem (8% - 9% decrease). The most significant changes were in the ether linked ferulates to lignin, both monomer and dehydrodiferulates (14% to 38% decrease). Other characteristics of the cell wall (lignin, neutral sugar composition) also showed modest changes. The change in total ferulates was modest, but led to improved animal performance. These findings suggest that relatively small changes can have a significant impact upon how well plant materials can be broken down and utilized by ruminants such as dairy cows.

Ferulic Acid Dehydrodimer and Dehydrotrimer Profiles of Distiller’s Dried Grains with Solubles from Different Cereal Species

Ferulic acid dehydrodimers (DFA) and dehydrotrimers (TriFA) ester-linked to plant cell wall polymers may cross-link not only cell wall polysaccharides but also other cell wall components including proteins and lignin, thus enhancing the rigidity and potentially affecting the enzymatic degradation of the plant cell wall. Corn, wheat, and mixed-cereal distiller's dried grains with solubles (DDGS) were investigated for composition of DFAs and TriFAs by reversed phase high-performance liquid chromatography with ultraviolet detection. Corn DDGS contained 5.3 and 5.9 times higher contents of total DFAs than wheat and mixed-cereal DDGS, respectively. Furthermore, the contents of total TriFAs were 5.7 and 6.3 times higher in corn DDGS than in wheat and mixed-cereal DDGS, respectively. In addition, both corn grains and corresponding DDGS had similar profiles of individual DFAs and TriFAs, indicating that ferulic acid cross-links in the corn cell wall are presumably not modified during fermentation and DDGS processing.

Comparing Corn Types for Differences in Cell Wall Characteristics and p-Coumaroylation of Lignin

This study was undertaken to compare cell wall characteristics including levels of p-coumarate (pCA) and lignin in corn (Zea mays L.) types. Five different types of corn, four commercial and Teosinte, were grown in the greenhouse in individual pots. For each corn type replicate stems were harvested at tassel emergence. Tissues for cell wall analysis were harvested from stems (separated into rind and pith tissues) and roots. Stem cell wall characteristics across the different corn types were similar for total neutral sugars, total uronosyls, lignin, and phenolic acids. However, the neutral sugar composition of root cell walls was markedly different, with high levels of galactose and arabinose. Levels of pCA in the different tissues ranged from 13.8 to 33.1 mg g(-1) of CW depending upon the type of tissue. There was no evidence that pCA was incorporated into cell walls attached to arabinoxylans. Lignin levels were similar within a given tissue, with pith ranging from 86.1 to 132.0 mg g(-1) of CW, rind from 178.4 to 236.6 mg g(-1) of CW, and roots from 216.5 to 242.6 mg g(-1) of CW. The higher values for lignins in root tissue may be due to suberin remaining in the acid-insoluble residue, forming Klason lignins. With the exception of root tissues, higher pCA levels accompanied higher lignin levels. This may indicate a potential role of pCA aiding lignin formation in corn cell walls during the lignification process.

Reduced genomic potential for secreted plant cell-wall-degrading enzymes in the ectomycorrhizal fungus Amanita bisporigera, based on the secretome of Trichoderma reesei

Based on the analysis of its genome sequence, the ectomycorrhizal (ECM) basidiomycetous fungus Laccaria bicolor was shown to be lacking many of the major classes of secreted enzymes that depolymerize plant cell wall polysaccharides. To test whether this is also a feature of other ECM fungi, we searched a survey genome database of Amanita bisporigera with the proteins found in the secretome of Trichoderma reesei (syn. Hypocrea jecorina), a biochemically well-characterized industrial fungus. Additional proteins were also used as queries to compensate for major groups of cell-wall-degrading enzymes lacking in the secretome of T. reesei and to substantiate conclusions drawn from the T. reesei collection. By MS/MS-based “shotgun” proteomics, 80 proteins were identified in culture filtrates of T. reesei strain RUTC30 grown on corn cell walls and in a commercial “cellulase” preparation, Spezyme CP. The two T. reesei enzyme preparations were qualitatively and quantitatively similar, the most striking difference being the lack of at least five major peptidases from the commercial enzyme mixture. Based on our analysis of A. bisporigera, this ECM fungus is deficient in many major classes of cell-wall-degrading enzymes, including both glycosyl hydrolases and carbohydrate esterases. By comparison, the genomes of the saprophytic basidiomycetes Coprinopsis cinerea and Galerina marginata (using a genome survey sequence approximately equivalent in depth to that of A. bisporigera) have, like T. reesei, a much more complete complement of cell-wall-degrading enzymes.

The Yeast Signal Sequence Trap Identifies Secreted Proteins of the Hemibiotrophic Corn PathogenColletotrichum graminicola

The hemibiotroph Colletotrichum graminicola is the causal agent of stem rot and leaf anthracnose on Zea mays. Following penetration of epidermal cells, the fungus enters a short biotrophic phase, followed by a destructive necrotrophic phase of pathogenesis. During both phases, secreted fungal proteins are supposed to determine progress and success of the infection. To identify genes encoding such proteins, we constructed a yeast signal sequence trap (YSST) cDNA-library from RNA extracted from mycelium grown in vitro on corn cell walls and leaf extract. Of the 103 identified unigenes, 50 showed significant similarities to genes with a reported function, 25 sequences were similar to genes without a known function, and 28 sequences showed no similarity to entries in the databases. Macroarray hybridization and quantitative reverse-transcriptase polymerase chain reaction confirmed that most genes identified by the YSST screen are expressed in planta. Other than some genes that were constantly expressed, a larger set showed peaks of transcript abundances at specific phases of pathogenesis. Another set exhibited biphasic expression with peaks at the biotrophic and necrotrophic phase. Transcript analyses of in vitro-grown cultures revealed that several of the genes identified by the YSST screen were induced by the addition of corn leaf components, indicating that host-derived factors may have mimicked the host milieu.

Characterization of p‐coumarate accumulation, p‐coumaroyl transferase, and cell wall changes during the development of corn stems

AbstractBACKGROUND: Developmental changes occur in corn (Zea mays L.) stems from cell initiation to fully mature cell types. During cell wall maturation the lignin is acylated with p‐coumarates (pCA). This work describes characterization studies of the p‐coumaroylation process in relation to corn stem development.RESULTS: Corn plants from three locations were harvested and tissues were analyzed from all nodes and even‐numbered internodes above soil line. Changes in carbohydrates reflect a shift to lignification at the expense of structural polysaccharide synthesis. Accumulation of pCA paralleled the incorporation of lignin while ferulate (FA) remained relatively constant as a proportion of the cell wall (5–7 g kg−1 CW). The p‐coumaroyl transferase (pCAT), which is responsible for attaching pCA to lignin monomers, displayed maximum levels of activity in the middle region of the stem (internodes 10–12, 2–3 nmol L−1 min−1 mg−1). The syringyl content as a proportion of the total lignin did not change significantly with cell wall maturation although there was a trend towards increased amounts of syringyl units in the more mature cell walls.CONCLUSIONS: Incorporation of pCA into corn cell walls not only mirrored lignification but the pCAT activity as well. Levels of pCAT activity may be an indicator of rapid lignification specifically for syringyl type lignin. Copyright © 2008 Society of Chemical Industry

Synergistic effects of cellulosomal xylanase and cellulases from Clostridium cellulovorans on plant cell wall degradation.

Plant cell walls are comprised of cellulose and hemicellulose and other polymers that are intertwined, and this complex structure presents a barrier to degradation by pure cellulases or hemicellulases. In this study, we determined the synergistic effects on corn cell wall degradation by the action of cellulosomal xylanase XynA and cellulosomal cellulases from Clostridium cellulovorans. XynA minicellulosomes and cellulase minicellulosomes were found to degrade corn cell walls synergistically but not purified substrates such as xylan and crystalline cellulose. The mixture of XynA and cellulases at a molar ratio of 1:2 showed the highest synergistic effect of 1.6 on corn cell wall degradation. The amounts both of xylooligosaccharides and cellooligosaccharides liberated from corn cell walls were increased by the synergistic action of XynA and cellulases. Although synergistic effects on corn cell wall degradation were found in simultaneous reactions with XynA and cellulases, no synergistic effects were observed in sequential reactions. The possible mechanism of synergism between XynA and cellulases is discussed.

Seed Protein Topology: Molecular Profiles

Corn, wheat, and soybean seed proteins were examined for potentially useful regularity, size, and polarity. Volume and amphiphilicity profiles evidence analogies to spider silk and human collagen, but seed proteins are 28% larger and 3.6 times more polar per residue. Wheat proteins are more uniform than silk; corn zeins less polar than collagen. A soybean cell wall protein is the most uniform (±5 A 3 ); a corn cell wall peptide is the smallest (92±9 A 3 ) and least polar (2±18) per residue. Selected parts appear to be more useful than whole molecules

Cloning and targeted gene disruption of XYL1, a beta 1,4-xylanase gene from the maize pathogen Cochliobolus carbonum.

The gene, XYL1, encoding the major extracellular endo-beta 1,4-xylanase from the maize pathogen Cochliobolus carbonum was cloned using a synthetic, degenerate oligonucleotide based on a tryptic fragment from the purified enzyme. The deduced product of XYL1 has a M(r) of 20,869 and a predicted pI of 9.1, in good agreement with the measured M(r) and pI of the purified enzyme. The XYL1 product has strong amino acid identity to seven endo-beta 1,4-xylanases from six prokaryotes but no obvious similarity to 10 other prokaryotic endoxylanases or a yeast endoxylanase. An internal fragment of the gene was used to create a specific xylanase mutant by transformation-mediated gene disruption via homologous recombination. Total extracellular xylanase activity in the mutant was reduced by 85-94%. When analyzed by cation exchange HPLC, culture filtrates of the mutant and wild type had identical protein profiles, but the mutant lacked the major peak of UV absorption corresponding to the major xylanase activity. Xylanase II activity was also missing in the mutant, but xylanase III activity was still present. The XYL1 mutant grew as well as the wild type on sucrose, on corn cell walls, and on xylan. The pathogenicity of the mutant was indistinguishable from the wild type, indicating that XYL1 is not required for pathogenicity.

Extracellular Enzymes Produced by Colletotrichum lindemuthianum and Helminthosporium maydis During Growth on Isolated Bean and Corn Cell Walls

Extracellular Enzymes Produced by Colletotrichum lindemuthianum and Helminthosporium maydis During Growth on Isolated Bean and Corn Cell Walls

Degradation of Corn Cell Walls by Extracellular Enzymes Produced by Helminthosporium maydis Race T

Degradation of Corn Cell Walls by Extracellular Enzymes Produced by Helminthosporium maydis Race T