Part:BBa_K2593004

1)LHyal gene ( leech HAase-encoding gene)
This part is a functional gene from Leech encoding a hyaluronidase called LHAase.

Biology

LHyal gene encodes LHAase (Mw=58kD), which is a hyaluronidases from the hyaluronate 3-glycanohydrolases sub-family of leech origin. Hyaluronidase (HAase) is a large family of glycoside hydrolases that predominantly degrades hyaluronic acid (HA), which is a polysaccharide composed of disaccharides unit of N-acetyl glucosamine and glucuronic acid polymerization. Based on substrate specificity and hydrolysis products, HAases are commonly grouped into three families: first group is hyaluronate lyases (EC 4.2.2.1, Streptococcus hyaluronate lyase), it’s commonly use to produce LHAase but has some potential risk on product. Second group is hyaluronate 4-glycanohydrolases (EC 3.2.1.35, Bovine testicular hyaluronidase, BTH) Commercial BTH has been widely used in clinical medicine, and its hydrolysis mechanism has been studied extensively. The disadvantages of the enzymatic production of specific or narrow-spectrum HA oligosaccharides by BTH include the limited source material (bovine testes), its considerably high price and the broad range of degradation products. Third group is hyaluronate 3-glycanohydrolases(EC 3.2.1.36, Leech HAase).Compared with BTH and Streptococcus hyaluronate lyase, leech HAase has higher substrate specificity and a narrow-spectrum of enzymatic products16,17. In particular, leech HAase is unable to degrade chondroitin or chondroitin sulfate compared to other HAase sources due to its strong substrate specificity. Although mammalian HAase has been widely used as a drug diffusion agent, such HAase activity is susceptible to heparin inhibition. Leech HAase activity, in theory, is not affected by heparin, and has more medical value in clinic and other medical aspects. In addition, the use of recombinant leech HAase does not pose any risk of animal cross-infection. Therefore, high-level production of recombinant leech HAase would be of great significance for both clinical medical treatment (such as surgery, ophthalmology and internal medicine) and producing narrow-spectrum HA oligosaccharides at the industrial scale.

Usage

Fig 1 . the LHyal gene was validated by using PCR primer pMA-LHyal-F and pMA-LHyal-R,the expected size is 520 bp

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 157