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Nonoichi, Japan

Ishikawa Prefectural University is a public university in the city of Nonoichi in Ishikawa Prefecture, Japan. The predecessor of the school was founded in 1971, and it was chartered as a university in 2005. Wikipedia.

The plastid genome of lettuce (Lactuca sativa L.) cv. Berkeley was site-specifically modified with the addition of three transgenes, which encoded β,β-carotenoid 3,3'-hydroxylase (CrtZ) and β,β-carotenoid 4,4'-ketolase (4,4'-oxygenase; CrtW) from a marine bacterium Brevundimonas sp. strain SD212, and isopentenyl diphosphate isomerase from a marine bacterium Paracoccus sp. strain N81106. Constructed transplastomic lettuce plants were able to grow on soil at a growth rate similar to that of non-transformed lettuce cv. Berkeley and generate flowers and seeds. The germination ratio of the lettuce transformants (T0) (98.8%) was higher than that of non-transformed lettuce (93.1 %). The transplastomic lettuce (T1) leaves produced the astaxanthin fatty acid (myristate or palmitate) diester (49.2% of total carotenoids), astaxanthin monoester (18.2%), and the free forms of astaxanthin (10.0%) and the other ketocarotenoids (17.5%), which indicated that artificial ketocarotenoids corresponded to 94.9% of total carotenoids (230 μg/g fresh weight). Native carotenoids were there lactucaxanthin (3.8%) and lutein (1.3 %) only. This is the first report to structurally identify the astaxanthin esters biosynthesized in transgenic or transplastomic plants producing astaxanthin. The singlet oxygen-quenching activity of the total carotenoids extracted from the transplastomic leaves was similar to that of astaxanthin (mostly esterified) from the green algae Haematococcus pluvialis. Source

Misawa N.,Ishikawa Prefectural University
Current Opinion in Biotechnology

Pathway engineering is to engineer biosynthetic pathways for compounds of interests in heterologous organisms such as microbes and higher plants, which has also been one of the most important fields in metabolic engineering and synthetic biology. This review focuses on pathway engineering researches for the production of functional isoprenoids containing monoterpenes, sesquiterpenes, diterpenes, and triterpenes as well as carotenoids and for the elucidation of relevant biosynthesis genes and enzymes, which have been performed in the last two years. As microbial hosts, Escherichia coli and Saccharomyces cerevisiae have often been employed, since they, specifically the former, are fully amenable to genetic manipulations with extensive molecular resources. Various crops have also been used as the hosts for engineering pathways of functional isoprenoids of the plant origin, particularly carotenoids. © 2011 Elsevier Ltd. Source

Ohki S.,Japan Advanced Institute of Science and Technology | Takeuchi M.,Japan Advanced Institute of Science and Technology | Mori M.,Ishikawa Prefectural University
Nature Communications

Stomatal development in plants is regulated by defensin-like secretory epidermal patterning factor (EPF) peptide hormones. Only one of these, stomagen, is a positive regulator, whereas EPF1, EPF2, and possibly others are negative regulators. Here we explore the structureĝ€"function relationships of EPFs, by integrating NMR and semi-in vitro stomagen experiments. We show that stomagen is composed of a loop and a scaffold containing three disulphide bonds. A mutant composed of the stomagen loop and the EPF2 scaffold positively regulates the stomatal density on Arabidopsis cotyledons. The reciprocal mutant composed of the EPF2 loop and the stomagen scaffold acts negatively. Deletion of the disulphide bond introduces unfolding and inactivity. Our results suggest that the loop confers the functional specificity of EPFs and that the scaffold is structurally required for their activity. This structural decomposition approach to elucidating the functional site could be adapted for the analysis of other cysteine-rich peptide families. © 2011 Macmillan Publishers Limited. All rights reserved. Source

Minami H.,Ishikawa Prefectural University
Bioscience, Biotechnology and Biochemistry

Higher plants produce diverse chemicals, including alkaloids, terpenoids, and phenolic compounds (phenylpropanoids and flavonoids) as secondary metabolites. These chemicals are widely used for human health and nutrition. Alkaloids, for example, are valued in medicine due to their high biological activities, but most of these metabolites accumulate at low levels in plant cells, resulting in poor extraction yields. Increasingly, attention is devoted to the production of plant metabolites by reconstructing plant biosynthetic pathways in microorganisms. This technology has been aided by advances in synthetic biology and metabolic engineering. Here, the review a fermentation platform for low-cost production of numerous alkaloids using bioengineered Escherichia coli and/or Saccharomyces cerevisiae. Source

Yamamoto K.,Ishikawa Prefectural University
Bioscience, Biotechnology and Biochemistry

This review describes the relationship between hetero-oligosaccharides and microorganisms. It is possible to prepare aminosugar nucleotides as donors for heterooligosaccharide synthesis with a combination of yeast fermentation and bacterial enzymes, and to use the product to test for a rare human blood group. We have isolated various glycosidases produced by microorganisms, mainly from soil, to elucidate the structure and function of hetero-oligosaccharides. Among them, a mold endoglycosidase was found to have specific transglycosylation activity in addition to hydrolysis activity, and we have used it to synthesize chemo-enzymatically various bioactive glycopeptides by the attachment of a hetero-oligosaccharide to a peptide. We found that lactic acid bacteria bound to a hetero-oligosaccharide on the intestinal tract cell surface in animals. We also analyzed the bifidobacterial hetero-oligosaccharidehydrolyzing enzymes involved in the degradation of mucin glycoprotein in the host intestinal tract and human milk oligosaccharides, and identified a specific saccharide that acted as a bifidobacteria growth factor. Source

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