Cereal seed storage protein synthesis: fundamental processes for recombinant protein production in cereal grains

2010 ◽  
Vol 8 (9) ◽  
pp. 939-953 ◽  
Author(s):  
Taiji Kawakatsu ◽  
Fumio Takaiwa
Keyword(s):  
Protein Synthesis ◽  
Recombinant Protein ◽  
Storage Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Seed Storage Protein ◽  
Seed Storage ◽  
Cereal Grains ◽  
Cereal Seed

scholarly journals The yeast eIF2 kinase Gcn2 facilitates H2O2-mediated feedback inhibition of both protein synthesis and ER oxidative folding during recombinant protein production

2021 ◽  
Author(s):  
Veronica Gast ◽  
Kate Campbell ◽  
Cecilia Picazo Campos ◽  
Martin Engqvist ◽  
Verena Siewers ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Recombinant Protein ◽  
Translation Initiation ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Feedback Inhibition ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Oxidative Folding ◽  
Amylase Production

AbstractRecombinant protein production is a known source of oxidative stress. Knowledge of which ROS are involved or the specific growth phase in which stress occurs however remains lacking. Using modern, hypersensitive genetic H2O2-specific probes, micro-cultivation and continuous measurements in batch culture, we observed H2O2 accumulation during and following the diauxic shift in engineered Saccharomyces cerevisiae, correlating with peak α-amylase production. In agreement with previous studies supporting a role of the translation initiation factor kinase Gcn2 in the response to H2O2, we find Gcn2-dependent phosphorylation of eIF2α to increase alongside translational attenuation in strains engineered to produce large amounts of α-amylase. Gcn2 removal significantly improved α-amylase production in two previously optimized high-producing strains, but not in the wild-type. Gcn2-deficiency furthermore reduced intracellular H2O2 levels and the unfolded protein response whilst expression of antioxidants and the ER disulfide isomerase PDI1 increased. These results suggest protein synthesis and ER oxidative folding to be coupled and subject to feedback inhibition by H2O2.ImportanceReactive oxygen species (ROS) accumulate during recombinant protein production both in yeast and Chinese hamster ovary cells, two of the most popular organisms used in the multi-million dollar protein production industry. Here we document increased H2O2 in the cytosol of yeast cells producing α-amylase. Since H2O2 predominantly targets the protein synthesis machinery and activates the translation initiation factor kinase Gcn2, we removed Gcn2, resulting in increased recombinant α-amylase production in two different previously engineered high-producing protein production strains. Removal of this negative feed-back loop thus represents a complementary strategy for improving recombinant protein production efforts currently used in yeast. Gcn2-deficiency also increased the expression of antioxidant genes and the ER-foldase PDI1, suggesting that protein synthesis and ER oxidative folding are linked and feed-back regulated via H2O2. Identification of additional components in this complex regulation may further improve protein production and contribute to the development of novel protein-based therapeutic strategies.

CONTROL OF SEED STORAGE PROTEIN SYNTHESIS IN DEVELOPING PISU

1981 ◽  
Vol 9 (2) ◽  
pp. 254P-254P ◽  
Author(s):  
R. R. D. CROY ◽  
J. A. GATEHOUSE ◽  
G. LYCETT ◽  
D. BOULTER
Keyword(s):  
Protein Synthesis ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Seed Storage

The role of mRNA and protein sorting in seed storage protein synthesis, transport, and deposition

2005 ◽  
Vol 83 (6) ◽  
pp. 728-737 ◽  
Author(s):  
Andrew J Crofts ◽  
Haruhiko Washida ◽  
Thomas W Okita ◽  
Mio Satoh ◽  
Masahiro Ogawa ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Storage Protein ◽  
Storage Proteins ◽  
Seed Storage Protein ◽  
Protein Localization ◽  
Protein Sorting ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Rna Localization ◽  
Endomembrane System

Rice synthesizes and accumulates high levels of 2 distinct classes of seed storage proteins and sorts them to separate intracellular compartments, making it an ideal model system for studying the mechanisms of storage protein synthesis, transport, and deposition. In rice, RNA localization dictates the initial site of storage protein synthesis on specific subdomains of the cortical endoplasmic reticulum (ER), and there is a direct relation between the RNA localization site and the final destination of the encoded protein within the endomembrane system. Current data support the existence of 3 parallel RNA localization pathways leading from the nucleus to the actively synthesizing cortical ER. Additional pathways may exist for the synthesis of cytoplasmic and nuclear-encoded proteins targeted to organelles, the latter located in a stratified arrangement in developing endosperm cells. The study of rice mutants, which accumulate unprocessed glutelin precursors, indicates that these multiple pathways prevent nonproductive interactions between different classes of storage proteins that would otherwise disrupt protein sorting. Indeed, it appears that the prevention of disruptive interactions between different classes of storage proteins plays a key role in their biosynthesis in rice. In addition to highlighting the unique features of the plant endomembrane system and describing the relation between RNA and protein localization, this minireview will attempt to address a number of questions raised by recent studies on these processes.Key words: mRNA localization, protein localization, endomembrane system, seed storage proteins, rice.

Analysis of Migrated Hydrogel Used for Breast Augmentation Revealed Prolamin (a Cereal Seed Storage Protein)

2011 ◽  
Vol 36 (1) ◽  
pp. 207-212
Author(s):  
Se Il Lee ◽  
Jung Kyu Han ◽  
Kun Hwang ◽  
Sowhey Park ◽  
Shin-A Jang
Keyword(s):  
Breast Augmentation ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Seed Storage ◽  
Cereal Seed

scholarly journals The yeast eIF2 kinase Gcn2 facilitates H 2 O 2 -mediated feedback inhibition of both protein synthesis and ER oxidative folding during recombinant protein production

2021 ◽  
Author(s):  
Veronica Gast ◽  
Kate Campbell ◽  
Cecilia Picazo ◽  
Martin Engqvist ◽  
Verena Siewers ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Feedback Inhibition ◽  
Translation Initiation Factor ◽  
Inhibitory Mechanism ◽  
Oxidative Folding ◽  
Amylase Production ◽  
Novel Protein

Recombinant protein production is a known source of oxidative stress. Knowledge of which ROS are involved or the specific growth phase in which stress occurs however remains lacking. Using modern, hypersensitive genetic H 2 O 2 -specific probes, micro-cultivation and continuous measurements in batch culture, we observed H 2 O 2 accumulation during and following the diauxic shift in engineered Saccharomyces cerevisiae , correlating with peak α-amylase production. In agreement with previous studies supporting a role of the translation initiation factor kinase Gcn2 in the response to H 2 O 2 , we find Gcn2-dependent phosphorylation of eIF2α to increase alongside translational attenuation in strains engineered to produce large amounts of α-amylase. Gcn2 removal significantly improved α-amylase production in two previously optimized high-producing strains, but not in the wild-type. Gcn2-deficiency furthermore reduced intracellular H 2 O 2 levels and the Hac1 splicing ratio whilst expression of antioxidants and the ER disulfide isomerase PDI1 increased. These results suggest protein synthesis and ER oxidative folding to be coupled and subject to feedback inhibition by H 2 O 2 . Importance Recombinant protein production is a multi-billion dollar industry. Optimizing the productivity of host cells is, therefore, of large interest. In several hosts oxidants are produced as an unwanted side product of recombinant protein production. The buildup of oxidants can result in intracellular stress responses which could compromise the productivity of the host cell. Here we document a novel protein synthesis inhibitory mechanism that is activated by the buildup of a specific oxidant (H 2 O 2 ) in the cytosol of yeast cells upon the production of recombinant proteins. At the center of this inhibitory mechanism lies the protein kinase Gcn2. By removing Gcn2 we observed a doubling of recombinant protein productivity in addition to reduced H 2 O 2 levels in the cytosol. By this study we want to raise awareness of this inhibitory mechanism in eukaryotic cells to further improve protein production and contribute to the development of novel protein-based therapeutic strategies.

scholarly journals A novel NAC family transcription factor SPR suppresses seed storage protein synthesis in wheat

2021 ◽  
Author(s):  
Lisha Shen ◽  
Guangbin Luo ◽  
Yanhong Song ◽  
Junyang Xu ◽  
JingJing Ji ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Protein Synthesis ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Seed Storage ◽  
Nac Family

scholarly journals Scanning probe microscopy studies of cereal seed storage protein structures

Scanning ◽  
1999 ◽  
Vol 21 (5) ◽  
pp. 293-298 ◽  
Author(s):  
Arthur S. Tatham ◽  
Neil H. Thomson ◽  
Terence J. McMaster ◽  
Andrew D. L. Humphris ◽  
Mervyn J. Miles ◽  
...  
Keyword(s):  
Scanning Probe Microscopy ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Protein Structures ◽  
Seed Storage ◽  
Scanning Probe ◽  
Probe Microscopy ◽  
Cereal Seed

scholarly journals Molecular cytogenetical and biochemical studies on some Lupinus species

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Hoda B. M. Ali ◽  
Samy A. A. Heiba
Keyword(s):  
Related Species ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Lupinus Albus ◽  
Seed Storage ◽  
Rrna Genes ◽  
Large Chromosome ◽  
45S Rdna ◽  
Lupinus Polyphyllus ◽  
Electrophoretic Patterns

Abstract Background Lupins are cultivated as human consumption grains and forage legumes. The chromosomes of lupins are too small to be karyotyped by conventional techniques, because they reveal a general lack of distinctive cytological features. In the current study, Fluorescence in situ Hybridization (FISH) was used to locate 5S and 45S ribosomal gene sites on the chromosomes of Lupinus albus ssp albus, Lupinus albus ssp graecus, Lupnus termis (all with 2n = 50), and Lupinus polyphyllus lindl var. polyphyllus (2n = 48), FISH together with seed storage protein electrophoretic patterns were used to find out the relationship among these species. Results The double-target FISH on the chromosomes of the studied species with rDNA probes revealed that the two types of rRNA genes are located on different chromosomes. The detected loci of rRNA genes partially reflected the taxonomical similarity among the two Lupinus albus subspecies and L. termis. Lupinus polyphyllus lindl var. polyphyllus was exception by having unique large chromosome mostly is covered by one signal of 45S rDNA, whereas its homologous chromosome seems to be normal-sized and have the other 45S rDNA locus. The similarity matrix among the Lupinus species as computed according to Jaccardʼs Coefficient from the SDS-PAGE, showed that L. albus ssp. Albus and L. albus ssp. Graecus are the most similar species (~ 97%), and then comes L. termis, and L. polyphyllus lindl var. polyphylus has been placed in separate clade and still the most related species to it among the studied species is L. termis (~ 70%). Conclusion It could be postulated from FISH and seed storage protein electrophoretic patterns that the relationships among the studied species is as follows, Lupinus albus ssp albus, is the most related species to Lupinus albus ssp graecus then comes Lupnus termis and Lupinus polyphyllus lindl var. polyphyllus at a distal position.

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