Table 3.

The advantages and disadvantages of main mutagenesis strategies.

Mutagenesis strategies	Different Forms	Advantages	Disadvantages	Refs
Random Mutagenesis	error-prone PCR, saturation mutagenesis, DNA shuffling	Huge library with various mutants	High proportion of non-relevant mutations, low efficiency	(84, 99, 100, 101, 102)
Target Mutagenesis	Ala scanning, site-directed mutation	More targeted, High efficiency	limited mutants, Point mutations usually have no effect.	(77, 78, 103, 104, 105, 106, 107, 108)
In silico mutagenesis	Protein analysis software combining different algorithms	Virtual huge library mutants; better structure and interaction analyze	Restrict by different algorithms; Lack of maturity and credibility	(73, 79, 109, 110, 111)