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Lysosomal Storage Disorders and Enzyme Replacement Therapy
Published in Peter Grunwald, Pharmaceutical Biocatalysis, 2020
Mucopolysaccharidosis IX (MPS IX or Natowicz Syndrome) is another extremely rare congenital disorder of which only a four cases have been reported up to 2015 in the relevant literature. MPS type IX is caused by the deficiency of enzyme hyaluronidase 1 (Hyal-1) which degrades hyaluronic acid (Kiykim et al., 2016) as schematically shown oppositely. In addition to its function as a passive structural molecule, e.g., as a component of articular cartilage and in tissue hydration, hyaluronan (HA) also acts as a signaling molecule by interacting with cell surface receptors (Vigetti et al., 2014), regulates cell proliferation a property depending on molecular weight and concentration of HA (Zhao et al., 2015), and cell migration (which is increased in combination with interleukin 6; Choi et al., 2014), as well as differentiation (e.g., embryonic stem cell (ESC) differentiation and embryogenesis (Choudhary et al., 2007; Simpson et al., 2016). In addition, HA plays a key role in regulating inflammation (Petrey and de la Motte, 2014) and is important in tumorigenesis (Mikami et al., 2018) and physiologic and pathological angiogenesis (Pardue et al., 2008). Hence, in clinical medicine, HA is used as a diagnostic marker for many diseases including cancer such as hepatocellular carcinoma, rheumatoid arthritis and liver pathologies (Kogan et al., 2007). Whereas in mammals GAG synthesis normally occurs in the Golgi apparatus, the site of HA biosynthesis is the cellular plasma membrane where it proceeds in presence of the three hyaluronan synthase isoenzymes (HAS1, HAS2, and HAS3) and uridine diphosphate (UDP) glucuronic acid (UDP-GlcUA) and UDP N-acetylglucosamine (UDP-GlcNAc) as building blocks (for a review of HA biosynthesis and signaling, see Vigetti et al., 2014).
Higher titer hyaluronic acid production in recombinant Lactococcus lactis
Published in Preparative Biochemistry and Biotechnology, 2018
Cansu Sunguroğlu, Dilber Ece Sezgin, Pınar Aytar Çelik, Ahmet Çabuk
The NICE system was used to drive expression of hasA gene under the control of the PnisA promoter in the expression plasmid pNZ8150. Recombinant L. lactis CES15 strain was cultured in M17G broth as a seed culture, until OD600 value reached 3. General protocol of nisin induction was followed as recommended by manufacturer.[29] Fresh medium was inoculated 1:25 (vol/vol) with this culture, cells were grown for ∼1 h in an OD600 of 0.3. Then, 0, 2, 5, 7.5, 10, and 12.5 ng/ml nisin were added into culture mediums and cells were incubated for further 3 hr to induce the expression of desired hyaluronan synthase enzyme. Optical density at 600 nm of each nisin concentration, at the end of the induction period were measured to be able to compare produced HA titers with each other.