title: Heterologous Production of Hyaluronic Acid in an ε-Poly-L-Lysine Producer, Streptomyces albulus.
abstract: Hyaluronic acid (HA) is used in a wide range of medical applications, where its performance and therapeutic efficacy are highly dependent on its molecular weight. In the microbial production of HA, it has been suggested that a high level of intracellular ATP enhances the productivity and molecular weight of HA. Here, we report on heterologous HA production in an ε-poly-l-lysine producer, Streptomyces albulus, which has the potential to generate ATP at high level. The hasA gene from Streptococcus zooepidemicus, which encodes HA synthase, was refactored and expressed under the control of a late-log growth phase-operating promoter. The expression of the refactored hasA gene, along with genes coding for UDP-glucose dehydrogenase, UDP-N-acetylglucosamine pyrophosphorylase, and UDP-glucose pyrophosphorylase, which are involved in HA precursor sugar biosynthesis, resulted in efficient production of HA in the 2.0 MDa range, which is greater than typical bacterial HA, demonstrating that a sufficient amount of ATP was provided to support the biosynthesis of the precursor sugars, which in turn promoted HA production. In addition, unlike in the case of streptococcal HA, S. albulus-derived HA was not cell associated. Based on these findings, our heterologous production system appears to have several advantages for practical HA production. We propose that the present system could be applicable to the heterologous production of a wide variety of molecules other than HA in the case their biosynthesis pathways require ATP in vivo.
authors: Yokohama Research Center, JNC Corporation, Yokohama, Kanagawa, Japan.
authors: Tomohiro
authors: T
authors: Yoshimura
authors: Yokohama Research Center, JNC Corporation, Yokohama, Kanagawa, Japan.
authors: Nobuyuki
authors: N
authors: Shibata
authors: Department of Bioscience, Fukui Prefectural University, Eiheiji-cho, Yoshida-gun, Fukui, Japan.
authors: Yoshimitsu
authors: Y
authors: Hamano
authors: Yokohama Research Center, JNC Corporation, Yokohama, Kanagawa, Japan kazuyay@kansai-u.ac.jp.
authors: Kazuya
authors: K
authors: Yamanaka
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D053898:Biosynthetic Pathways / Q000235:genetics*; D006820:Hyaluronic Acid / Q000494:pharmacology*; D060847:Metabolic Engineering; D011107:Polylysine / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D011107:Polylysine; D000255:Adenosine Triphosphate; D006820:Hyaluronic Acid