Alginate, a polysaccharide found in brown algae, plays a role of great
importance for the world both in marine ecosystems and in industrial
applications. Nevertheless, the true potential of marine polysaccharides
is limited in a way due to its disorganized branching and macromolecule
figure, which leads to its low solubility and high solution viscosity.
Besides, its capability to be easily absorbed by the body is weakened to
a certain extent, leading to inconvenience to put it into further use,
such as systemic therapies. The need to find its replacement is then
highlighted. It’s noticed that with the glycosidic bonds between the
β-D-mannuronic acid(M) and α-L-guluronic acid(G) residues cleaved away
from alginate, a better material referred to as alginate
oligosaccharide(AOS) is generated[1]. Studies have shown its anti-tumor,
anti-virus, and anti-inflammatory effects. Moreover, easier diffusion
and absorption features are shown on it as a result of its smaller size.
It also functions in biological activities like signaling and biofilms
penetration[2,3]. To make AOS available, the enzyme that specifically
helps with the breakdown of alginate is necessary. Performing such
unique ability to degrade alginate, alginate lyase deserves further
improvement to become more efficient when it comes to facilitate the
process. Thus, we planned to improve the enzyme activity of alginate
lyase, through which we found the error-prone PCR method was possibly to
achieve our goal.
Error-prone PCR is an evolution of standard PCR
technology. It is a technique that makes DNA more prone to mismatches
during the amplification process, hence it is also known as mismatch PCR
or error-prone PCR. Error-prone PCR typically utilizes low-fidelity Taq
DNA polymerase and alters some components of the PCR reaction system to
reduce the fidelity of DNA replication during the PCR process,
increasing the mutation rate and thus obtaining different DNA sequences
or genes [4,5].
