title: Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: cytological studies.
abstract: Extraction of hyaluronan from animals or microbial fermentation has risks including contamination with pathogens and microbial toxins. In this work, tobacco cultured-cells (BY-2) were successfully transformed with a chloroviral hyaluronan synthase (cvHAS) gene to produce hyaluronan. Cytological studies revealed accumulation of HA on the cells, and also in subcellular fractions (protoplasts, miniplasts, vacuoplasts, and vacuoles). Transgenic BY-2 cells harboring a vSPO-cvHAS construct containing the vacuolar targeting signal of sporamin connected to the N-terminus of cvHAS accumulated significant amounts of HA in vacuoles. These results suggested that cvHAS successfully functions on the vacuolar membrane and synthesizes/transports HA into vacuoles. Efficient synthesis of HA using this system provides a new method for practical production of HA.
authors: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima 739-8530, Japan.
authors: Numfon
authors: N
authors: Rakkhumkaew
authors: Shigeo
authors: S
authors: Shibatani
authors: Takeru
authors: T
authors: Kawasaki
authors: Makoto
authors: M
authors: Fujie
authors: Takashi
authors: T
authors: Yamada
mesh_terms: D001483:Base Sequence; D002473:Cell Wall; D002478:Cells, Cultured; D017931:DNA Primers; D004798:Enzymes / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000235:genetics*; D000076002:Hyaluronan Synthases; D006820:Hyaluronic Acid / Q000096:biosynthesis* / Q000378:metabolism; D015388:Organelles / Q000378:metabolism; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D013347:Subcellular Fractions / Q000201:enzymology; D014026:Nicotiana / Q000166:cytology / Q000201:enzymology*
chemical_list: D017931:DNA Primers; D004798:Enzymes; D006820:Hyaluronic Acid; D014453:Glucuronosyltransferase; D000076002:Hyaluronan Synthases