Method of increasing the catalytic activity of a glycanase...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/56 (2006.01) A23K 1/165 (2006.01) A23K 1/18 (2006.01) C12N 1/21 (2006.01) C12N 9/24 (2006.01) C12N 15/74 (2006.01)

Patent

CA 2136784

A method for increasing the catalytic activity of a glycanase having two or more independent catalytic domains by isolating the catalytic domains from the intact enzyme is described. If the catalytic domains in the intact enzyme are linked by a proline/hydroxyamino acid-rich linker sequence, the catalytic domains can be isolated and activated by cleavage within or adjacent to the linker sequence with a protease. The catalytic domains can also be isolated, whether or not they are linked by a hydroxyamino acid-rich linker sequence, by expressing the DNA sequences encoding each domain in separate transformed microbial hosts. An isolated and purified low-activity xylanase, denoted XynC, and which is obtained from Fibrobacter succinogenes is also described. XynC contains two independant active catalytic domains separated by a serine rich linker region. The enzyme has a mass of 63 kDa and an activity of 6 U/mg (µmol/min/mg) protein. The subcloned domains expressed separatedly exhibit 55 to 116 fold higher activity than the intact enzyme has a specific activity of approximately 6 U/mg protein and a mass of 63 kDa and consisting of two independently active catalytic domains linked by a hydroxyamino acid-rich linker sequence. XynC can be activated by treatment of the intact enzyme with a protease. The utilization of feed by domesticated animals and poultry can be improved by admixing the feed with XynC and a catalytic quantity of protease as the XynC is activated by proteases indigenous to the digestive tract of the animal.

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